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MedWorm: Microbiology



MedWorm.com provides a medical RSS filtering service. Over 7000 RSS medical sources are combined and output via different filters. This feed contains the latest news and research in Microbiology



Last Build Date: Mon, 28 Mar 2016 07:44:02 +0100

 



The MIT lab flushing out a city’s secrets

Sun, 27 Mar 2016 08:00:11 +0100

Researchers in Massachusetts are looking at ways to tackle public health issues by delving into the sewers. Luckily, a robot does all the dirty work…Here in this small room within the Massachusetts Institute of Technology in Cambridge, MA, I am making the acquaintance of Luigi. As explorers go, he lacks charisma – not for him a winning smile, witty catchphrase and firm handshake. But then, Luigi isn’t your typical pioneer. He’s a robot. And it isn’t curios he collects. It’s sewage.At first take, it seems an unlikely subject for Luigi’s creators – the Senseable City Lab – to embrace. Along twisting corridors, sleek black panels showcase the group’s off-the-wall ideas: LED-clad micro helicopters, location-tracked trash, the Copenhagen wheel – a motorised hub for bicycle...

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Prevalence of Leishmania infantum and co-infections in stray cats in northern Italy.

Sun, 27 Mar 2016 05:20:02 +0100

Authors: Spada E, Canzi I, Baggiani L, Perego R, Vitale F, Migliazzo A, Proverbio D Abstract Stray cats in the city of Milan, Italy, were tested for Leishmania infantum and other selected infections. Twenty-seven cats (30.0%) were seroreactive by indirect fluorescent antibody test (IFAT), with an antibody titer of 1:40 for 16 (17.7%) cats and 1:80 (cut-off for feline L. infantum infection) for 11 (12.2%) cats. One blood (1.1%) and one popliteal lymph node (1.1%) sample tested positive by real-time polymerase chain reaction; no oculoconjunctival swabs tested positive. Feline immunodeficiency virus, feline leukemia virus, and feline coronavirus (FCoV) seroprevalence determined by enzyme-linked immunosorbent assay was 6.1, 6.1, and 39.0%, respectively. Toxoplasma gondii, Bartonella he...



Pheno- and genotypic characterization of Pasteurella multocida isolated from cats, dogs and rabbits from Brazil.

Sun, 27 Mar 2016 05:20:02 +0100

This study aimed to characterize the genetic diversity of P. multocida isolated from dogs, cats and rabbits, and to evaluate their antimicrobial susceptibility profiles. A total of 620 animals were studied; 51 were positive for P. multocida and 92 strains were isolated. 60.9% of the strains belonged to the capsular type A, while the remaining were classified as non-typeable. The hgbA, ptfA, sodC, tadD and hsf2 genes were more frequent among the rabbit strains. Sulfonamides and cotrimoxazole presented the highest resistance rate, followed by erythromycin. PFGE clustered strains according to host species. Our results indicate that P. multocida from companion animals carry several virulence factors and are resistant to antimicrobials commonly used in human and veterinary medicine. PMID: ...



Cathepsin K inhibition renders equine bone marrow nucleated cells hypo-responsive to LPS and unmethylated CpG stimulation in vitro.

Sun, 27 Mar 2016 05:20:02 +0100

In conclusion, CatK inhibition in horses did affect BMNCs other than mature osteoclasts rendering them hypo-responsive to both TLR4- and TLR9-induced inflammation, predicting a proteolytic activity for CatK within the MyD88 pathway and/or the following proteolytic events required for the cytokines secretion. PMID: 27012920 [PubMed - in process] (Source: Comparative immunology, microbiology and infectious diseases.)



Antimicrobial resistance in faecal Escherichia coli isolates from farmed red deer and wild small mammals. Detection of a multiresistant E. coli producing extended-spectrum beta-lactamase.

Sun, 27 Mar 2016 05:20:02 +0100

Authors: Alonso CA, González-Barrio D, Tenorio C, Ruiz-Fons F, Torres C Abstract Eighty-nine Escherichia coli isolates recovered from faeces of red deer and small mammals, cohabiting the same area, were analyzed to determine the prevalence and mechanisms of antimicrobial resistance and molecular typing. Antimicrobial resistance was detected in 6.7% of isolates, with resistances to tetracycline and quinolones being the most common. An E. coli strain carrying blaCTX-M-1 as well as other antibiotic resistant genes included in an unusual class 1 integron (Intl1-dfrA16-blaPSE-1-aadA2-cmlA1-aadA1-qacH-IS440-sul3-orf1-mef(B)Δ-IS26) was isolated from a deer. The blaCTX-M-1 gene was transferred by conjugation and transconjugants also acquired an IncN plasmid. This strain was typed as ST22...



Clinical isolates of Acinetobacter baumannii from a Portuguese hospital: PFGE characterization, antibiotic susceptibility and biofilm-forming ability.

Sun, 27 Mar 2016 05:20:02 +0100

Authors: Duarte A, Ferreira S, Almeida S, Domingues FC Abstract Acinetobacter baumannii is an emerging pathogen associated with nosocomial infections that in addition has shown an increasing resistance to antibiotics. In this work the genetic diversity of A. baumannii isolates from a Portuguese hospital, their antibiotic resistance profiles and ability to form biofilms was studied. Seventy-nine clinical A. baumannii isolates were characterized by pulsed-field gel electrophoresis (PFGE) with 9 different PFGE profiles being obtained. Concerning the antimicrobial susceptibility, all A. baumannii isolates were resistant to 12 of the 17 tested antibiotics and classified as multidrug-resistant (MDR). In addition, 74.7% of the isolates showed biofilm formation ability, however no statisti...

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Molecular evidence of vector-borne pathogens in dogs and cats and their ectoparasites in Algiers, Algeria.

Sun, 27 Mar 2016 05:20:02 +0100

Authors: Bessas A, Leulmi H, Bitam I, Zaidi S, Ait-Oudhia K, Raoult D, Parola P Abstract In Algeria, only limited information is currently available on the prevalence of emergent canine and feline vector-borne diseases. The aim of the present work was to detect by qPCR vector-associated bacteria in stray dogs and cats and their ectoparasites from Algiers. 18/117 (15.38%) dogs and 2/107 (1.87%) cats were positive for at least one vector-borne agent. Coxiella burnetii and Bartonella henselae were identified in 1/117 (0.85%) dog individually. Ehrlichia canis DNA was detected in 17/117 (14.52%) dogs. 1/107 (0.93%) cat was positive to C. burnetii and another 1/107 (0.93%) to B. henselae. DNA of Rickettsia massiliae, Rickettsia conorii and E. canis was detected in Rhipicephalus sanguineu...



Eyeworm infections in dogs and in a human patient in Serbia: A One Health approach is needed.

Sun, 27 Mar 2016 05:20:02 +0100

We describe the first case of human thelaziosis in Serbia, along with two cases in dogs from the same area. A One Health approach, based on cooperation amongst veterinarians and physicians, is strongly advised for this emerging infection in order to assess the risk for and prevent of the zoonotic infection. PMID: 27012916 [PubMed - in process] (Source: Comparative immunology, microbiology and infectious diseases.)



An evaluation of ELISA using recombinant Brucella abortus bacterioferritin (Bfr) for bovine brucellosis.

Sun, 27 Mar 2016 05:20:02 +0100

In this study, cloning and expression of Brucella abortus bacterioferritin (Bfr) was accomplished by PCR amplification into an expression vector system, and purification of a recombinant B. abortus Bfr (rBfr). The immunogenicity of rBfr was confirmed by Western blot with Brucella-positive bovine serum. To determine whether rBfr has a potential benefit for use in the serodiagnosis of bovine brucellosis, rBfr-based ELISA was performed. Interestingly, rBfr was able to detect anti-Brucella antibodies in positive sera in a dependent manner of TAT values but did not show an immunoreaction with negative samples. Particularly, average OD492 values at the lowest, medium and highest TAT titer levels were 1.4, 2.2 and 2.6-fold increase compared with the cutoff value, respectively. The accuracy, speci...



Faecal virome of red foxes from peri-urban areas.

Sun, 27 Mar 2016 05:20:02 +0100

In this study, we evaluated the faecal virome of juvenile and adult foxes from peri-urban areas in central Croatia. The dominating mammalian viruses were fox picobirnavirus and parvovirus. The highest number of viral reads (N=1412) was attributed to a new fox circovirus and complete viral genome was de novo assembled from the high-throughput sequencing data. Fox circovirus is highly similar to dog circoviruses identified in diseased dogs in USA and Italy, and to a recently discovered circovirus of foxes with neurologic disease from the United Kingdom. Our fox picobirnavirus was more closely related to the porcine and human picobirnaviruses than to known fox picobirnaviruses. PMID: 27012914 [PubMed - in process] (Source: Comparative immunology, microbiology and infectious diseases.)



Quillaja brasiliensis saponins induce robust humoral and cellular responses in a bovine viral diarrhea virus vaccine in mice.

Sun, 27 Mar 2016 05:20:02 +0100

Authors: Cibulski SP, Silveira F, Mourglia-Ettlin G, Teixeira TF, Dos Santos HF, Yendo AC, de Costa F, Fett-Neto AG, Gosmann G, Roehe PM Abstract A saponin fraction extracted from Quillaja brasiliensis leaves (QB-90) and a semi-purified aqueous extract (AE) were evaluated as adjuvants in a bovine viral diarrhea virus (BVDV) vaccine in mice. Animals were immunized on days 0 and 14 with antigen plus either QB-90 or AE or an oil-adjuvanted vaccine. Two-weeks after boosting, antibodies were measured by ELISA; cellular immunity was evaluated by DTH, lymphoproliferation, cytokine release and single cell IFN-γ production. Serum anti-BVDV IgG, IgG1 and IgG2b were significantly increased in QB-90- and AE-adjuvanted vaccines. A robust DTH response, increased splenocyte proliferation, Th1-ty...

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[Can the urine dipstick be used in the diagnosis of urinary bacterial colonizations in a preoperative urological assessment?]

Sun, 27 Mar 2016 00:33:05 +0100

CONCLUSION: In spite of a good negative predictive value linked to a low prevalence of colonized patients (5%), the low sensitivity of the urine dipstick test entails a non-negligible number of false negatives. Its use as a single test of preoperative screening would expose colonized patients to the prospect of an operation, which seems to be unacceptable for some of them, notably endoscopic ones. LEVEL OF EVIDENCE: 4. PMID: 27012836 [PubMed - as supplied by publisher] (Source: Progres en Urologie)



Acute Pneumonia

Sat, 26 Mar 2016 07:44:23 +0100

Acute pneumonia is an active infection of the lungs that results when an individual at risk gets exposed to a particular microbiological pathogen. Acute pneumonia is the leading cause of death in the United States that is attributable to an infection. The risk factors, pathogenesis, and microbiological organisms involved differ if the pneumonia develops in the community versus health care-associated environment. The development of concise and comprehensive guidelines has led to an improvement in the management of the problem. However, the emergence of multidrug-resistant organisms and the increase in the percentage of elderly population keep mortality risk very substantial. (Source: Critical Care Nursing Quarterly)



Evaluation of Ag containing hydroxyapatite coatings to the Candida albicans infection

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 26 March 2016 Source:Journal of Microbiological Methods Author(s): S. Ciuca, M. Badea, E. Pozna, I. Pana, A. Kiss, L. Floroian, A. Semenescu, C.M. Cotrut, M. Moga, A. Vladescu In this research work, the synthesis of Ag doped hydroxyapatite coatings for dental or orthopedic implants was performed. The main goal was to determine the influence of Ag content on the roughness and antimicrobial performance of the prepared thin films. The films were deposited on Ti6Al4V alloy by means of RF magnetron sputtering. Those coatings were characterized by X-ray diffraction (XRD) and 3D surface profilometry. The antifungal activity after 1 and 7days of culture was evaluated in the presence of Candida albicans (ATCC — 10231). The increase of Ag content incr...



Optimisation of a droplet digital PCR assay for the diagnosis of Schistosoma japonicum infection: A duplex approach with DNA binding dye chemistry

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 26 March 2016 Source:Journal of Microbiological Methods Author(s): Kosala G. Weerakoon, Catherine A. Gordon, Geoffrey N. Gobert, Pengfei Cai, Donald P. McManus Schistosomiasis is a chronically debilitating helminth infection with a significant socio-economic and public health impact. Accurate diagnostics play a pivotal role in achieving current schistosomiasis control and elimination goals. However, many of the current diagnostics procedures, which rely on detection of schistosome eggs, have major limitations including lack of accuracy and the inability to detect pre-patent infections. DNA-based detection methods provide a viable alternative to the current tests commonly used for schistosomiasis diagnosis. Here we describe the optimisation of a nov...

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Multiplex real-time PCR assay for the detection of extended-spectrum β-lactamase and carbapenemase genes using melting curve analysis

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 25 March 2016 Source:Journal of Microbiological Methods Author(s): Prashant Singh, Yvonne Pfeifer, Azlin Mustapha Real-time PCR melt curve assays for the detection of β-lactamase, extended-spectrum β-lactamase and carbapenemase genes in Gram-negative bacteria were developed. Two multiplex real-time PCR melt curve assays were developed for the detection of ten common β-lactamase genes: bla KPC-like, bla OXA-48-like, bla NDM-like, bla VIM-like, bla IMP-like, bla CTX-M-1+2-group, bla CMY-like, bla ACC-like, bla SHV-like and bla TEM-like. The assays were evaluated using 25 bacterial strains and 31 DNA samples (total n=56) comprising different Enterobacteriaceae genera and Pseudomonas spp. These strains were previously characterized at five research in...



Development of simple sequence repeat (SSR) markers for discrimination among isolates of Fusarium proliferatum

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 25 March 2016 Source:Journal of Microbiological Methods Author(s): I. Moncrief, C. Garzon, S. Marek, J. Stack, A. Gamliel, P. Garrido, F. Proaño, M. Gard, H. Dehne, J. Fletcher The plant pathogen Fusarium proliferatum has a wide host range and occurs worldwide. Many isolates of the fungus produce mycotoxins in plant tissues, which, if ingested, can cause harm to animals and humans. In 2008, an outbreak of salmon blotch of onions, caused by F. proliferatum, was detected in southern Israel. The source and distribution of the fungus in Israel were unknown. Inter-simple sequence repeats (ISSR) were used to identify repetitive motifs present in seven isolates of F. proliferatum from Israel, Germany and Austria. ISSR repeat motifs were, used to dev...



Cpe1786/IscR of Clostridium perfringens represses expression of genes involved in Fe-S cluster biogenesis

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 26 March 2016 Source:Research in Microbiology Author(s): Gaelle André, Elise Haudecoeur, Emmanuelle Courtois, Marc Monot, Bruno Dupuy, Dmitry A. Rodionov, Isabelle Martin-Verstraete Cpe1786 of Clostridium perfringens is an Rrf2-type regulator containing the three-cysteine residues coordinating a Fe-S in IscR, the repressor controlling Fe-S homeostasis in enterobacteria. The cpe1786 gene formed an operon with iscSU involved in Fe-S biogenesis and tmrU. This operon was transcribed from a σA-dependent promoter. We showed that in the heterologous host Bacillus subtilis, Cpe1786, renamed IscRCp, negatively controlled its own transcription. We constructed an iscR mutant in C. perfringens. We then compared the expression profile of strain 13 and of th...



Metagenomes provide valuable comparative information on soil microeukaryotes

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 26 March 2016 Source:Research in Microbiology Author(s): Samuel Jacquiod, Jonas Stenbæk, Susana S. Santos, Anne Winding, Søren J. Sørensen, Anders Priemé Despite the critical ecological roles of microeukaryotes in terrestrial ecosystems, most descriptive studies of soil microbes published so far focused only on specific groups. Meanwhile, the fast development of metagenome sequencing leads to considerable data accumulation in public repositories, providing microbiologists with substantial amounts of accessible information. We took advantage of public metagenomes in order to investigate microeukaryote communities in a well characterized grassland soil. The data gathered allowed the evaluation of several factors impacting the community structure...



Sensitivity of IgG ELISA for diagnosing cysticercosis in high risk group in and around Aligarh District of Uttar Pradesh, India

Fri, 25 Mar 2016 23:00:00 +0100

Conclusions Cysticercosis is a health problem in and around Aligarh with higher risk in males. Seroprevalence of cysticercosis in high risk community of Aligarh is 7.4%. (Source: Asian Pacific Journal of Tropical Disease)

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Barriers to uptake of antimicrobial advice in a UK hospital: a qualitative study

Fri, 25 Mar 2016 23:00:00 +0100

Conclusions This study illustrates the importance of examining relations between ID/CM and non-ID/CM clinicians in the hospital context, indicating that AMS models that focus exclusively on delivering advice rather than managing interprofessional relationships may be limited in their capacity to optimize antibiotic use. AMS and, specifically, antibiotic approval systems would likely be more effective if they incorporated time and resources for fostering and maintaining professional relationships. (Source: Journal of Hospital Infection)



Utility of multiplex real-time PCR in the diagnosis of extrapulmonary tuberculosis

Fri, 25 Mar 2016 23:00:00 +0100

Conclusion PCR can play an important role in rapid and accurate diagnosis of extrapulmonary tuberculosis. IS6110 alone is an effective target in our part of the country. (Source: The Brazilian Journal of Infectious Diseases)



Propionibacterium prosthetic joint infection: experience from a retrospective database analysis

Fri, 25 Mar 2016 23:00:00 +0100

Conclusion Patients with persistent postoperative pain and/or loosening of implants should be screened for PJI with low-virulent organisms such as Propionibacterium, including. (Source: European Journal of Orthopaedic Surgery and Traumatology)



Therapeutic effect of Pseudomonas aeruginosa phage YH30 on mink hemorrhagic pneumonia

Fri, 25 Mar 2016 23:00:00 +0100

The objective of this study was to investigate the potential of using phages as a therapy against hemorrhagic pneumonia in mink. A broad-host-range phage from the Podoviridae family, YH30, was isolated using the mink-originating P. aeruginosa (serotype G) D7 strain as a host. The genome of YH30 was 72,192bp (54.92% G+C), contained 86 open reading frames and lacked regions encoding known virulence factors, integration-related proteins or antibiotic resistance determinants. These characteristics make YH30 eligible for use in phage therapy. The results of a curative treatment experiment demonstrated that a single intranasal administration of YH30 was sufficient to cure hemorrhagic pneumonia in mink. The mean colony count of P. aeruginosa in the blood and lung of YH30-protected mink was less t...



Chloroquine inhibits Rhodococcus equi replication in murine and foal alveolar macrophages by iron-starvation

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 26 March 2016 Source:Veterinary Microbiology Author(s): Leticia T. Gressler, Angela I. Bordin, Cole M. McQueen, Noah D. Cohen, Agueda Castagna de Vargas Rhodococcus equi preferentially infects macrophages causing pyogranulomatous pneumonia in young foals. Both the vapA and rhbC genes are up-regulated in an iron (Fe)-deprived environment, such as that found within macrophages. Chloroquine (CQ) is a drug widely used against malaria that suppresses the intracellular availability of Fe in eukaryotic cells. The main objective of this study was to evaluate the ability of CQ to inhibit replication of virulent R. equi within murine (J774A.1) and foal alveolar macrophages (AMs) and to verify whether the mechanism of inhibition could be Fe-deprivation-depend...

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Evolution of the nasopharyngeal microbiota of beef cattle from weaning to 40days after arrival at a feedlot

Fri, 25 Mar 2016 23:00:00 +0100

The objective was to describe the evolution of the nasopharyngeal microbiota of beef cattle from weaning to 40days after arrival at a feedlot. Deep nasal swabs (DNS) from 30 Angus-cross steers were collected at weaning, on arrival at a feedlot, and at day 40 after arrival. The DNA was extracted from DNS and the hypervariable region V3 of the 16S rRNA gene was amplified and sequenced (Illumina MiSeq platform). Nasopharyngeal microbiota underwent a profound evolution from weaning to arrival at the feedlot and from arrival to day 40, with the abundance of 92 Operational Taxonomic Units (OTUs) significantly changing over time. Mycoplasma (M. dispar and M. bovirhinis) was the most abundant genus in the nasopharynx, accounting for 53% of the total bacterial population. Because an evolving bacter...



Persistence of nasal colonisation with methicillin resistant Staphylococcus aureus CC398 among participants of veterinary conferences and occurrence among their household members: a prospective cohort study, Germany 2008 to 2014

Fri, 25 Mar 2016 23:00:00 +0100

Publication date: Available online 26 March 2016 Source:Veterinary Microbiology Author(s): Jan Walter, Werner Espelage, Cornelia Adlhoch, Christiane Cuny, Susanne Schink, Andreas Jansen, Wolfgang Witte, Tim Eckmanns, Julia Hermes There are only few data on the persistence and transmission of methicillin resistant Staphylococcus aureus (MRSA) of the clonal complex (CC) 398 among veterinarians and their household members. We therefore investigated the long-term colonisation with MRSA CC398 among participants of veterinary conferences in Germany in 2008/2009 and their household members. Forty-five initially MRSA CC398 positive and 180 initially MRSA CC398 negative conference participants were included in a longitudinal study. These persons and their household members were tested fo...



Fish oils against Burkholderia and Pseudomonas aeruginosa: in vitro efficacy and their therapeutic and prophylactic effects on infected Galleria mellonella larvae

Fri, 25 Mar 2016 21:22:18 +0100

ConclusionThe fish oils, particularly in the free fatty acid form, are active in killing Burkholderia and P. aeruginosa.This article is protected by copyright. All rights reserved. (Source: Journal of Applied Microbiology)



Association of Cell-adhesion Activities with Virulence in Shiga toxin-producing Escherichia coli O103:H2.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Kobayashi N, Maeda E, Saito S, Furukawa I, Ohnishi T, Watanabe M, Terajima J, Hara-Kudo Y Abstract The characteristics of 11 strains of Stx1-producing and Stx2-non-producing STEC O103:H2 were analyzed to investigate the differences in virulence in a single serotype of Shiga toxin (Stx) -producing Escherichia coli (STEC). Differences in the cell-adhesion activity to Caco-2 cells were observed among the strains. The activity of the one strain, isolated from a patient with hemolytic uremic syndrome was 4-20-fold higher than those of the other strains. Although the strains with high cell-adhesion activity showed high expressions of eae, espB, espD, and tir in the locus of enterocyte effacement related with cell-adhesion, those were not specific for this strain. In addition, ...

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Fate of Escherichia coli O157 Cells Inoculated into Lightly Pickled Chinese Cabbage during Processing, Storage and Incubation in Artificial Gastric Juice.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Inatsu Y, Ohata Y, Ananchaipattana C, Latiful Bari M, Hosotani Y, Kawasaki S Abstract Fate of Escherichia coli O157 cells was evaluated when inoculated into each step after production of lightly pickled Chinese cabbage. The efficacy of surface sterilization by 100 mg/L of chlorine water for 10 min on raw leaves (6.0 log CFU/g) was 2.2 log CFU/g reduction. No meaningful change of the population of E. coli O157 (3.5 log CFU/g to 1.5 log MPN/g) contaminated into 19 kinds of products was observed. These results indicated the difficulty of estimating the viable count of the cells between contaminated on farms and further processing and storage steps. The population of E. coli O157 (3 log CFU/g to 1 log MPN/g) inoculated into the Chinese cabbage products was reduced less than 0....



Identification of Cereulide-Producing Bacillus cereus by Nucleic Acid Chromatography and Reverse Transcription Real-Time PCR.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Ueda S, Yamaguchi M, Eguchi K, Iwase M Abstract RNA extracts were analyzed with a nucleic acid sequence-based amplification (NASBA) - nucleic acid chromatography and a reverse transcription-quantitative PCR assay (RT-qPCR) based on the TaqMan probe for identification of cereulide-producing Bacillus cereus. All 100 emetic B. cereus strains were found to give positive results, but 50 diarrheal B. cereus strains and other bacterial species showed negative results in the NASBA-chromatography. That is, the assay could selectively identify the emetic strains among B. cereus strains. Also, the B. cereus contents of more than 10(7) cfu/ml were required for the identification of the cereulide-producing strains in this assay. In qRT-PCR assays, all 100 emetic type strains of B. cere...



Low-Pressure Plasma Application for the Inactivation of the Seed-borne Pathogen Xanthomonas campestris.

Fri, 25 Mar 2016 17:05:02 +0100

In this study, the effectiveness of low-pressure plasma treatment in the inactivation of the seed-borne plant pathogenic bacterium, Xanthomonas campestris, inoculated on cruciferous seeds, was evaluated. The highest inactivation effect was observed when the treatment voltage and argon gas flow rate were 5.5 kV and 0.5 L/min, respectively. The viable cell number of X. campestris was 6.6 log cfu/seed before plasma treatment, and decreased by 3.9 log after 5 min of treatment and by 6.6 log after 40 min. Ethidium monoazide treatment and quantitative real-time PCR results indicated that both the cell membrane and target DNA region were damaged following 5 min of plasma treatment. Although both heat and ozone were generated during the plasma treatment, the contribution of both factors to the ina...



Isolation and Characterization of a Purple Non-Sulfur Photosynthetic Bacterium Rhodopseudomonas faecalis Strain A from Swine Sewage Wastewater.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Wei H, Okunishi S, Yoshikawa T, Kamei Y, Maeda H Abstract A purple non-sulfur photosynthetic bacterium (PNSB), PSB Strain A was isolated from swine sewage wastewater. Phylogenetic analysis revealed that PSB Strain A was most closely related to Rhodopseudomonas faecalis. Growth of the isolate under anaerobic-light conditions with a variety of carbon sources was investigated. Both PSB Strain A and the standard strain showed good growth with acetic acid, propionic acid, and n-butyric acid at a concentration of 20 mM. At the high concentration of 200 mM, PSB Strain A showed better growth in pyruvate, acetate, propionate, succinate and malate. By applying PSB Strain A to treat swine sewage wastewater, the concentration of VFAs, which were acetic acid and propionic acid, decreas...



Relationship between Virucidal Efficacy and Free Iodine Concentration of Povidone-Iodine in Buffer Solution.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Wada H, Nojima Y, Ogawa S, Hayashi N, Sugiyama N, Kajiura T, Ueda T, Morimoto S, Yokota K Abstract Povidone-iodine solutions prepared to various concentrations (0.01, 0.1, 1 and 10%) with 0.2M phosphate buffer (pH 7.0) (PVP-I PB) were analyzed to determine their free iodine concentrations using membrane permeation cells, and their inactivation effects on three viruses (influenza A virus, poliovirus type 1 and adenovirus type 3) were examined. The free iodine concentrations in the 0.01-10% PVP-I PB were determined to be 1.84, 4.88, 1.58 and 0.17 ppm (approximate values), respectively, with the maximum obtained for the 0.1% solution. The virucidal efficacy of these PVP-I PB against poliovirus type 1 and adenovirus type 3 was found to be generally dependent on free iodine con...

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Effects of Modified Atmosphere Packaging, Food Life Extenders and Temperature on the Shelf Life of Ready-Made Dishes.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Sato J, Maenishi T, Saito Y, Masuda T, Kadotani N, Kozakai H, Ito M Abstract The combined effect of several microbial control factors including gas barrier of containers, modified atmosphere packaging, food life extenders and storage temperature was discussed in order to determine the possibility for improving the shelf life for hamburger steak and deepfried chicken, representative ready-made dishes sold at convenience stores in Japan. Multiple measures including cold storage were effective in improving the shelf life of ready-made dishes. It was also suggested that storage tests for ready-made dishes should be conducted at 10℃, a practical temperature, to confirm the storable period, as well as at 15℃, an adequate abuse temperature, to confirm the effects of various m...



Inactivation of Bacterial Spore, Endotoxin, Lipid A, Normal Prion and Abnormal Prion by Exposures to Several Sorts of Gases Plasma.

Fri, 25 Mar 2016 17:05:02 +0100

Authors: Shintani H Abstract This review discusses the application of several sorts of non-equilibrium gas plasma discharges for sterilization and disinfection treatments against spores or bioburden on/in the healthcare products or biological indicators. The basic properties of electrical discharges are briefly reviewed and thereafter the paper discusses the interactions of gas plasma with several sorts of biological systems such as bacteria, bacterial spores, endotoxins, lipid A and normal and abnormal prion proteins. PMID: 27009504 [PubMed - in process] (Source: Biocontrol Science)



Epidemiology of hospital-acquired pneumonia: Results of a Central European multicenter, prospective, observational study compared with data from the European region.

Fri, 25 Mar 2016 15:35:02 +0100

CONCLUSIONS: The present study was the first to collect multicenter data on the epidemiology of HAP in the Central European Region, with respect to the incidence of etiologic agents causing HAP. It was concerned with relationships between 30-day patient mortality and the type of HAP, etiologic agent and adequacy of initial empirical antibiotic therapy. PMID: 27003315 [PubMed - as supplied by publisher] (Source: Biomedical Papers of the Medical Faculty of the Univ Palacky Olomouc Czech Repub)



Stripped-Down Synthetic Organism Has Smallest Genome Yet

Fri, 25 Mar 2016 14:41:58 +0100

This study is definitely trying to understand a minimal basis of life," said Venter. But the researchers said that even with such a simple organism, that understanding remained elusive. They noted that even though their organism has so few genes, they were still uncertain about the function of nearly a third of them, even after more than five years of work. The researchers predicted their work would yield practical applications in developing new medicines, biochemicals, biofuels and in agriculture. "Our long-term vision has been to design and build synthetic organisms on demand where you can add in specific functions and predict what the outcome is going to be," said Daniel Gibson, vice president for DNA technologies at Synthetic Genomics Inc, the company handling commercial applications f...



Archaeal Community Changes Associated with Cultivation of Amazon Forest Soil with Oil Palm.

Fri, 25 Mar 2016 14:21:02 +0100

This study compared soil archaeal communities of the Amazon forest with that of an adjacent area under oil palm cultivation by 16S ribosomal RNA gene pyrosequencing. Species richness and diversity were greater in native forest soil than in the oil palm-cultivated area, and 130 OTUs (13.7%) were shared between these areas. Among the classified sequences, Thaumarchaeota were predominant in the native forest, whereas Euryarchaeota were predominant in the oil palm-cultivated area. Archaeal species diversity was 1.7 times higher in the native forest soil, according to the Simpson diversity index, and the Chao1 index showed that richness was five times higher in the native forest soil. A phylogenetic tree of unclassified Thaumarchaeota sequences showed that most of the OTUs belong to Miscellaneo...

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Strategies to Block Bacterial Pathogenesis by Interference with Motility and Chemotaxis.

Fri, 25 Mar 2016 14:18:01 +0100

Authors: Erhardt M Abstract Infections by motile, pathogenic bacteria, such as Campylobacter species, Clostridium species, Escherichia coli, Helicobacter pylori, Listeria monocytogenes, Neisseria gonorrhoeae, Pseudomonas aeruginosa, Salmonella species, Vibrio cholerae, and Yersinia species, represent a severe economic and health problem worldwide. Of special importance in this context is the increasing emergence and spread of multidrug-resistant bacteria. Due to the shortage of effective antibiotics for the treatment of infections caused by multidrug-resistant, pathogenic bacteria, the targeting of novel, virulence-relevant factors constitutes a promising, alternative approach. Bacteria have evolved distinct motility structures for movement across surfaces and in aqueous environmen...



Editorial Board.

Fri, 25 Mar 2016 08:08:01 +0100

Authors: PMID: 27001962 [PubMed - in process] (Source: Applied and Environmental Microbiology)



Articles of Significant Interest Selected from This Issue by the Editors.

Fri, 25 Mar 2016 08:08:01 +0100

Authors: PMID: 27001961 [PubMed - in process] (Source: Applied and Environmental Microbiology)

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Botulism in waterbirds: Mortality rates and new insights into how it spreads

Fri, 25 Mar 2016 04:00:00 +0100

(American Society for Microbiology) Outbreaks of botulism killed large percentages of waterbirds inhabiting a wetland in Spain. The botulinum toxin's spread may have been abetted by an invasive species of water snail which frequently carries the toxin-producing bacterium, Clostridium botulinum, and which is well adapted to wetlands polluted by sewage. Global warming will likely increase outbreaks, said corresponding author Rafael Mateo, Ph.D. The research was published March 25 in Applied and Environmental Microbiology, a journal of the American Society for Microbiology. (Source: EurekAlert! - Biology)



Induced Bacterial Cross-Resistance toward Host Antimicrobial Peptides: A Worrying Phenomenon

Fri, 25 Mar 2016 02:24:10 +0100

Osmel Fleitas, Octávio L. Franco (Source: Frontiers in Microbiology)



Management of septic shock and severe infections in migrants and returning travelers requiring critical care

Fri, 25 Mar 2016 02:23:13 +0100

Abstract During the past decade, global human movement created a virtually “borderless world”. Consequently, the developed world is facing “forgotten” and now imported infectious diseases. Many infections are observed upon travel and migration, and the clinical spectrum is diverse, ranging from asymptomatic infection to severe septic shock. The severity of infection depends on the etiology and timeliness of diagnosis. While assessing the etiology of severe infection in travelers and migrants, it is important to acquire a detailed clinical history; geography, dates of travel, places visited, type of transportation, lay-overs and intermediate stops, potential exposure to exotic diseases, and activities that were undertaken during travelling and prophylaxis and vaccines eith...

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Outbreak of NDM-1-producing Klebsiella pneumoniae ST76 and ST37 isolates in neonates

Fri, 25 Mar 2016 02:23:13 +0100

Abstract The purpose of this study was to investigate the epidemiological characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP) in Shanghai Children’s Hospital in China. Twenty-two non-duplicate CRKP strains were collected from pediatric patients between March and June in 2014. Antimicrobial susceptibility testing was conducted by the agar dilution method. Beta-lactamases were characterized by polymerase chain reaction (PCR) and DNA sequencing. The transferability of bla NDM-1 was investigated by conjugation experiment. The plasmids bearing antibiotic resistance genes were characterized by S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) and Southern hybridization. Clonal relatedness was evaluated by PFGE and multilocus sequence typing (MLST)...



A survey to identify barriers of implementing an antibiotic checklist

Fri, 25 Mar 2016 02:23:13 +0100

Abstract A checklist is an effective implementation tool, but addressing barriers that might impact on the effectiveness of its use is crucial. In this paper, we explore barriers to the uptake of an antibiotic checklist that aims to improve antibiotic use in daily hospital care. We performed an online questionnaire survey among medical specialists and residents with various professional backgrounds from nine Dutch hospitals. The questionnaire consisted of 23 statements on anticipated barriers hindering the uptake of the checklist. Furthermore, it gave the possibility to add comments. We included 219 completed questionnaires (122 medical specialists and 97 residents) in our descriptive analysis. The top six anticipated barriers included: (1) lack of expectation of improvement of a...



Time to positivity of blood culture and its prognostic value in bloodstream infection

Fri, 25 Mar 2016 02:23:12 +0100

Abstract The purpose of this study was to investigate the relationship between the time to positivity (TTP) of blood cultures and outcome in patients with bloodstream infections (BSIs). Between January 1st, 2011 and December 31st, 2013, the blood cultures of inpatients with BSI or catheter-related BSI were collected at Peking University Third Hospital. The TTP of different isolates was analyzed, and the relationship between the TTP of isolates and outcome of patients with Enterobacter BSI was retrospectively analyzed. We analyzed the TTP of 886 isolates. Escherichia coli has the shortest (11.97 ± 10.06 h) and Candida has the longest first TTP (61.62 ± 42.77 h). 68.01 % of isolates reached positivity within 24 h and 88.33 % within 48 h. Over 90 % of E. coli isolat...



Epidemiological and clinical features for cefepime heteroresistant Escherichia coli infections in Southwest China

Fri, 25 Mar 2016 02:23:12 +0100

This study aimed to report the characteristics of invasive E. coli with FEP-PHR traits and further to investigate the predisposing factors for its acquisition. A retrospective case–control study was conducted in a teaching hospital in Southwest China. A total of 319 successive and non-duplicate E. coli strains were isolated from blood and other sterile body fluids between July 2011 and August 2013. Among the seventy (70/319, 21.9 %) isolates harboring FEP-PHR traits, 30 (42.9 %) isolates were isolated from blood, 14 (20.0 %) isolates were isolated from bile, and 13 (18.6 %) isolates were isolated from drainage. FEP-PHR isolates were verified by population analysis profile (PAP) assays. Male gender, receipt of total parenteral nutrition, cephalosporins exposure, and production of exte...



In vivo development of daptomycin resistance in vancomycin-susceptible methicillin-resistant Staphylococcus aureus severe infections previously treated with glycopeptides

Fri, 25 Mar 2016 02:23:12 +0100

Abstract Our aim was to describe the clinical and microbiological features of four cases of severe vancomycin-susceptible methicillin-resistant Staphylococcus aureus (MRSA) infections in which the vancomycin non-susceptibility development and daptomycin resistance occurred under therapy with teicoplanin (three cases) and daptomycin switched to vancomycin (one case). Clinical data were retrospectively reviewed. On nine clinical epidemiologically unrelated daptomycin-susceptible (DAP-S) and daptomycin-resistant (DAP-R) MRSA, we performed: (i) DAP-VAN-TEC-CFX-RIF minimum inhibitory concentrations (MICs); (ii) glycopeptide resistance detection (GRD) by δ-hemolysis; (iii) glycopeptide population analysis; (iv) molecular characterization by PFGE-MLST-SCCmec-agr-typing; (v) rpoB and mp...

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Bacterial vaginosis, aerobic vaginitis, vaginal inflammation and major Pap smear abnormalities

Fri, 25 Mar 2016 02:23:12 +0100

Abstract The purpose of this investigation was to evaluate the impact of the vaginal milieu on the presence of abnormal Pap smears and a positive human papilloma virus (HPV) test. A cross-sectional study was conducted between June 2014 and May 2015, evaluating the vaginal discharge by fresh wet mount microscopy and comparing these data with Pap smear findings. Wet mount slides were scored for bacterial vaginosis (BV), aerobic vaginitis (AV), presence of Candida and Trichomonas vaginalis. Cytologic evaluation was done on all Pap smears according to the Bethesda criteria. The cobas© HPV Test (Roche) was performed for HPV detection. A total of 622 cases were evaluated. The mean age of the patients was 41.6 ± 10.65 years (range 21–75). Eighty-three women (13.3 %) had a cytol...



Improved detection of deeply invasive candidiasis with DNA aptamers specific binding to (1→3)-β-D-glucans from Candida albicans

Fri, 25 Mar 2016 02:23:12 +0100

In this study, we sought to screen high-affinity DNA aptamers that specifically bound to (1→3)-β-D-glucans from cell wall of Candida albicans using a systematic evolution of ligands by exponential enrichment (SELEX) technique, and further evaluate the diagnostic potential for invasive or disseminated candidiasis with selected aptamers. (1→3)-β-D-glucans was purified from Candida albicans, and two single DNA aptamers (designated as AU1 and AD1) were selected. Analysis of dissociation constants and binding domains further revealed that these two selected single DNA aptamers (AU1 and AD1) showed high binding affinity (AD1: Kd = 79.76 nM, AD1: Kd = 103.7 nM) and did not bind to the same domain of (1→3)-β-D-glucans. Next, we further detected (1→3)-β-D-glucans in serum samp...



Diversity of ribosomal 16S DNA‐ and RNA‐based bacterial community in an office building drinking water system

Fri, 25 Mar 2016 02:22:56 +0100

ConclusionsMicrobial communities differed between water and biofilms, between cold and hot water systems, locations in the building and between water rRNA and rDNA libraries, as shown by clear clusters in principal component analysis (PcoA). By using the rRNA method, we found that not all bacterial community members were active (e.g. Legionella spp.) while other members showed increased activity in some locations; e.g. Pseudomonas spp. in hot water circulations’ biofilm and order Rhizobiales and Limnohabitans spp. in stagnated locations’ water and biofilm. Significance and Impact of the StudyrRNA‐based methods may be better than rDNA‐based methods for evaluating human health implications as rRNA methods can be used to describe the active bacterial fraction. This study indicates tha...



NapM, a new nucleoid‐associated protein, broadly regulates gene expression and affects mycobacterial resistance to anti‐tuberculosis drugs

Fri, 25 Mar 2016 02:22:43 +0100

This article is protected by copyright. All rights reserved. NapM represents a new nucleoid‐associated protein and is conserved in all of the sequenced mycobacterial genomes. It recognizes DNA in a length‐dependent but sequence‐independent manner. NapM globally regulates the expression of more than 150 genes and the resistance of Mycobacterium smegmatis to two anti‐tuberculosis drugs. (Source: Molecular Microbiology)



Are humans the initial source of canine mange?

Fri, 25 Mar 2016 00:00:00 +0100

Scabies, or mange as it is called in animals, is an ectoparasitic contagious infestation caused by the mite Sarcoptes scabiei. Sarcoptic mange is an important veterinary disease leading to significant morbidity a... (Source: Parasites and Vectors)

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Zika outbreak in Americas could be down to one plane passenger

Thu, 24 Mar 2016 23:14:14 +0100

Study of virus’s evolution shows strains in Americas share a single ancestor, and suggest it could have arrived during the 2013 Fifa Confederations CupThe outbreak of Zika virus in the Americas could be down to just one passenger that stepped off a plane in 2013.New research suggests that the virus could have arrived in Brazil more than a year before it was first reported in March 2015, scotching the notion that it was brought to Brazil during the 2014 World Cup, or arrived during the Va’a World Sprint Championship canoe race. Continue reading... (Source: Guardian Unlimited Science)



Use of an attenuated live Salmonella Typhimurium vaccine on three breeding pig units: A longitudinal observational field study

Thu, 24 Mar 2016 23:00:00 +0100

Publication date: June 2016 Source:Comparative Immunology, Microbiology and Infectious Diseases, Volume 46 Author(s): R. Davies, R.J Gosling, A.D. Wales, R.P. Smith The study examined the effects of a licensed live Salmonella Typhimurium vaccine, administered to sows and gilts on three commercial pig units experiencing clinical salmonellosis associated with S. Typhimurium or its monophasic variant. After vaccination, clinical salmonellosis resolved and shedding of S. Typhimurium declined markedly and persistently on all breeding or breeding-finishing units, during the one- to two-year monitoring period. On two finishing units supplied in part by one of the vaccinated herds, pigs from the vaccinated herd were less likely to shed Salmonella than those from non-vaccinating herds, and Sa...



Discovering novel enzymes by Functional screening of plurigenomic libraries from alga-associated Flavobacteriia and Gammaproteobacteria

Thu, 24 Mar 2016 23:00:00 +0100

Publication date: Available online 24 March 2016 Source:Microbiological Research Author(s): Marjolaine Martin, Marie Vandermies, Coline Joyeux, Renée Martin, Tristan Barbeyron, Gurvan Michel, Micheline Vandenbol Alga-associated microorganisms, in the context of their numerous interactions with the host and the complexity of the marine environment, are known to produce diverse hydrolytic enzymes with original biochemistry. We recently isolated several macroalgal-polysaccharide-degrading bacteria from the surface of the brown alga Ascophyllum nodosum. These active isolates belong to two classes: the Flavobacteriia and the Gammaproteobacteria. In the present study, we constructed two “plurigenomic” (with multiple bacterial genomes) libraries with the 5 most interesting isolates ...



C. kronotskyensis SLH Glycoside Hydrolases [Enzymology]

Thu, 24 Mar 2016 23:00:00 +0100

The genome of the extremely thermophilic bacterium Caldicellulosiruptor kronotskyensis encodes 19 surface layer (S-layer) homology (SLH) domain-containing proteins, the most in any Caldicellulosiruptor species genome sequenced to date. These SLH proteins include five glycoside hydrolases (GHs) and one polysaccharide lyase, the genes for which were transcribed at high levels during growth on plant biomass. The largest GH identified so far in this genus, Calkro_0111 (2,435 amino acids), is completely unique to C. kronotskyensis and contains SLH domains. Calkro_0111 was produced recombinantly in Escherichia coli as two pieces, containing the GH16 and GH55 domains, respectively, as well as putative binding and spacer domains. These displayed endo- and exoglucanase activity on the β-1,3-1,6-gl...



Ad19a E3/49K Trafficking and Proteolytic Processing [Microbiology]

Thu, 24 Mar 2016 23:00:00 +0100

The E3 transcription unit of human species C adenoviruses (Ads) encodes immunomodulatory proteins that mediate direct protection of infected cells. Recently, we described a novel immunomodulatory function for E3/49K, an E3 protein uniquely expressed by species D Ads. E3/49K of Ad19a/Ad64, a serotype that causes epidemic keratokonjunctivitis, is synthesized as a highly glycosylated type I transmembrane protein that is subsequently cleaved, resulting in secretion of its large ectodomain (sec49K). sec49K binds to CD45 on leukocytes, impairing activation and functions of natural killer cells and T cells. E3/49K is localized in the Golgi/trans-Golgi network (TGN), in the early endosomes, and on the plasma membrane, yet the cellular compartment where E3/49K is cleaved and the protease involved r...

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GGCT Activated by Eukaryotic Thioredoxins [Cell Biology]

Thu, 24 Mar 2016 23:00:00 +0100

The plant pathogenic bacterium Ralstonia solanacearum injects more than 70 effector proteins (virulence factors) into the host plant cells via the needle-like structure of a type III secretion system. The type III secretion system effector proteins manipulate host regulatory networks to suppress defense responses with diverse molecular activities. Uncovering the molecular function of these effectors is essential for a mechanistic understanding of R. solanacearum pathogenicity. However, few of the effectors from R. solanacearum have been functionally characterized, and their plant targets remain largely unknown. Here, we show that the ChaC domain-containing effector RipAY/RSp1022 from R. solanacearum exhibits γ-glutamyl cyclotransferase (GGCT) activity to degrade the major intracellular re...



Inner Membrane Complex of Type IV Pilus Assembly Systems [Microbiology]

Thu, 24 Mar 2016 23:00:00 +0100

Type IV pili (T4P) are ubiquitous bacterial cell surface structures, involved in processes such as twitching motility, biofilm formation, bacteriophage infection, surface attachment, virulence, and natural transformation. T4P are assembled by machinery that can be divided into the outer membrane pore complex, the alignment complex that connects components in the inner and outer membrane, and the motor complex in the inner membrane and cytoplasm. Here, we characterize the inner membrane platform protein PilC, the cytosolic assembly ATPase PilB of the motor complex, and the cytosolic nucleotide-binding protein PilM of the alignment complex of the T4P machinery of Myxococcus xanthus. PilC was purified as a dimer and reconstituted into liposomes. PilB was isolated as a monomer and bound ATP in...



Role of Pyruvate Kinase in M. tuberculosis [Metabolism]

Thu, 24 Mar 2016 23:00:00 +0100

Mycobacterium tuberculosis (Mtb) displays a high degree of metabolic plasticity to adapt to challenging host environments. Genetic evidence suggests that Mtb relies mainly on fatty acid catabolism in the host. However, Mtb also maintains a functional glycolytic pathway and its role in the cellular metabolism of Mtb has yet to be understood. Pyruvate kinase catalyzes the last and rate-limiting step in glycolysis and the Mtb genome harbors one putative pyruvate kinase (pykA, Rv1617). Here we show that pykA encodes an active pyruvate kinase that is allosterically activated by glucose 6-phosphate (Glc-6-P) and adenosine monophosphate (AMP). Deletion of pykA prevents Mtb growth in the presence of fermentable carbon sources and has a cidal effect in the presence of glucose that correlates with e...



Editorial Board [Masthead]

Thu, 24 Mar 2016 23:00:00 +0100

(Source: Journal of Clinical Microbiology)



Photo Quiz: A Man with Fever and Cough [Photo Quiz]

Thu, 24 Mar 2016 23:00:00 +0100

(Source: Journal of Clinical Microbiology)



The Brief Case: A Reactive HIV Rapid Antibody Test in a Pregnant Woman [The Brief Case]

Thu, 24 Mar 2016 23:00:00 +0100

(Source: Journal of Clinical Microbiology)



Point-Counterpoint: The FDA Has a Role in Regulation of Laboratory-Developed Tests [Point-Counterpoint]

Thu, 24 Mar 2016 23:00:00 +0100

Since the Food and Drug Administration (FDA) released its draft guidance on the regulation of laboratory-developed tests (LDTs) in October 2014, there has been a flurry of responses from commercial and hospital-based laboratory directors, clinicians, professional organizations, and diagnostic companies. The FDA defines an LDT as an "in vitro diagnostic device that is intended for clinical use and is designed, manufactured, and used within a single laboratory." The draft guidance outlines a risk-based approach, with oversight of high-risk and moderate-risk tests being phased in over 9 years. High-risk tests would be regulated first and require premarket approval. Subsequently, moderate-risk tests would require a 510(k) premarket submission to the FDA and low-risk tests would need only to be...

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HIV Whole-Genome Sequencing Now: Answering Still-Open Questions [Commentaries]

Thu, 24 Mar 2016 23:00:00 +0100

Diversity, evolution, and epidemiology of HIV are directly relevant to HIV transmission and pathogenesis; hence, they play a key role in antiretroviral treatment and vaccine design. Global HIV whole-genome sequencing would provide a treasure chest of data to answer many questions still open in these fields. An article by Berg et al. in this issue of the Journal of Clinical Microbiology describes a universal strategy to amplify and sequence heterogeneous HIV whole genomes (M. G. Berg, J. Yamaguchi, E. Alessandri-Gradt, R. W. Tell, J.-C. Plantier, and C. A. Brennan, J Clin Microbiol 54:868–882, 2016, http://dx.doi.org/10.1128/JCM.02479-15). (Source: Journal of Clinical Microbiology)



Identification of Low-Level Vancomycin Resistance in Staphylococcus aureus in the Era of Informatics [Commentaries]

Thu, 24 Mar 2016 23:00:00 +0100

Vancomycin-intermediate Staphylococcus aureus (VISA) and heteroresistant VISA (hVISA) are pathogens for which accurate antimicrobial susceptibility testing (AST) would rule out standard treatment with vancomycin. Unfortunately, AST for vancomycin is relatively slow and standard methods are unable to reliably detect VISA and hVISA. An article in this issue (C. A. Mather, B. J. Werth, S. Sivagnanam, D. J. SenGupta, and S. M. Butler-Wu, J Clin Microbiol 54:883–890, 2016, doi:http://dx.doi.org/10.1128/JCM.02428-15) describes a rapid whole-cell matrix-assisted laser desorption ionization–time of flight proxy susceptibility method that highlights current innovations and challenges with rapid AST, VISA/hVISA identification, and clinical bioinformatics. (Source: Journal of Clinical Mic...



Impact of CLSI Breakpoint Changes on Microbiology Laboratories and Antimicrobial Stewardship Programs [Commentaries]

Thu, 24 Mar 2016 23:00:00 +0100

In 2010, the Clinical and Laboratory Standards Institute (CLSI) lowered the MIC breakpoints for many beta-lactam antibiotics to enhance detection of known resistance among Enterobacteriaceae. The decision to implement these new breakpoints, including the changes announced in both 2010 and 2014, can have a significant impact on both microbiology laboratories and antimicrobial stewardship programs. In this commentary, we discuss the changes and how implementation of these updated CLSI breakpoints requires partnership between antimicrobial stewardship programs and the microbiology laboratory, including data on the impact that the changes had on antibiotic usage at our own institution. (Source: Journal of Clinical Microbiology)



Interferon Gamma Release Assays for Latent Tuberculosis: What Are the Sources of Variability? [Minireviews]

Thu, 24 Mar 2016 23:00:00 +0100

Interferon gamma release assays (IGRAs) are blood-based tests intended for diagnosis of latent tuberculosis infection (LTBI). IGRAs offer logistical advantages and are supposed to offer improved specificity over the tuberculin skin test (TST). However, recent serial testing studies of low-risk individuals have revealed higher false conversion rates with IGRAs than with TST. Reproducibility studies have identified various sources of variability that contribute to nonreproducible results. Sources of variability can be broadly classified as preanalytical, analytical, postanalytical, manufacturing, and immunological. In this minireview, we summarize known sources of variability and their impact on IGRA results. We also provide recommendations on how to minimize sources of IGRA variability. (So...



Consolidating and Exploring Antibiotic Resistance Gene Data Resources [Minireviews]

Thu, 24 Mar 2016 23:00:00 +0100

The unrestricted use of antibiotics has resulted in rapid acquisition of antibiotic resistance (AR) and spread of multidrug-resistant (MDR) bacterial pathogens. With the advent of next-generation sequencing technologies and their application in understanding MDR pathogen dynamics, it has become imperative to unify AR gene data resources for easy accessibility for researchers. However, due to the absence of a centralized platform for AR gene resources, availability, consistency, and accuracy of information vary considerably across different databases. In this article, we explore existing AR gene data resources in order to make them more visible to the clinical microbiology community, to identify their limitations, and to propose potential solutions. (Source: Journal of Clinical Microbiology...

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Zika Virus: Diagnostics for an Emerging Pandemic Threat [Minireviews]

Thu, 24 Mar 2016 23:00:00 +0100

Zika virus (ZIKV) is an Aedes mosquito-borne flavivirus that emerged in Brazil in 2015 and then rapidly spread throughout the tropical and subtropical Americas. Based on clinical criteria alone, ZIKV cannot be reliably distinguished from infections with other pathogens that cause an undifferentiated systemic febrile illness, including infections with two common arboviruses, dengue virus and chikungunya virus. This minireview details the methods that are available to diagnose ZIKV infection. (Source: Journal of Clinical Microbiology)



A Pan-HIV Strategy for Complete Genome Sequencing [Virology]

Thu, 24 Mar 2016 23:00:00 +0100

Molecular surveillance is essential to monitor HIV diversity and track emerging strains. We have developed a universal library preparation method (HIV-SMART [i.e., switching mechanism at 5' end of RNA transcript]) for next-generation sequencing that harnesses the specificity of HIV-directed priming to enable full genome characterization of all HIV-1 groups (M, N, O, and P) and HIV-2. Broad application of the HIV-SMART approach was demonstrated using a panel of diverse cell-cultured virus isolates. HIV-1 non-subtype B-infected clinical specimens from Cameroon were then used to optimize the protocol to sequence directly from plasma. When multiplexing 8 or more libraries per MiSeq run, full genome coverage at a median ~2,000x depth was routinely obtained for either sample type. The method rep...



Rapid Detection of Vancomycin-Intermediate Staphylococcus aureus by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Vancomycin is the standard of care for the treatment of invasive methicillin-resistant Staphylococcus aureus (MRSA) infections. Infections with vancomycin-nonsusceptible MRSA, including vancomycin-intermediate S. aureus (VISA) and heterogeneous VISA (hVISA), are clinically challenging and are associated with poor patient outcomes. The identification of VISA in the clinical laboratory depends on standard susceptibility testing, which takes at least 24 h to complete after isolate subculture, whereas hVISA is not routinely detected in clinical labs. We therefore sought to determine whether VISA and hVISA can be differentiated from vancomycin-susceptible S. aureus (VSSA) using the spectra produced by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF ...



Variable-Number Tandem-Repeat Analysis of Respiratory and Household Water Biofilm Isolates of "Mycobacterium avium subsp. hominissuis" with Establishment of a PCR Database [Mycobacteriology and Aerobic Actinomycetes]

Thu, 24 Mar 2016 23:00:00 +0100

"Mycobacterium avium subsp. hominissuis" is an important cause of pulmonary disease. It is acquired from environmental sources, but there is no methodology for large population studies. We evaluated the potential of variable-number tandem-repeat (VNTR) analysis. Clinical and household biofilm M. avium isolates underwent molecular identification. Testing for IS901 was done to separate M. avium subsp. avium from M. avium subsp. hominissuis. VNTR types were defined using VNTR loci, and subtyping was performed using 3' hsp65 and internal transcribed spacer (ITS) sequencing. Forty-nine VNTR types and eight subtypes of M. avium subsp. hominissuis (IS901 negative) were identified among 416 isolates of M. avium from 121 patients and 80 biofilm sites. Of those types, 67% were found only among patie...



Novel Assays for Measurement of Total Cell-Associated HIV-1 DNA and RNA [Virology]

Thu, 24 Mar 2016 23:00:00 +0100

Although a number of PCR-based quantitative assays for measuring HIV-1 persistence during suppressive antiretroviral therapy (ART) have been reported, a simple, sensitive, reproducible method is needed for application to large clinical trials. We developed novel quantitative PCR assays for cell-associated (CA) HIV-1 DNA and RNA, targeting a highly conserved region in HIV-1 pol, with sensitivities of 3 to 5 copies/1 million cells. We evaluated the performance characteristics of the assays using peripheral blood mononuclear cells (PBMCs) from 5 viremic patients and 20 patients receiving effective ART. Total and resting CD4+ T cells were isolated from a subset of patients and tested for comparison with PBMCs. The estimated standard deviations including interassay variability and intra-assay v...

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MTBDRplus and MTBDRsl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis [Mycobacteriology and Aerobic Actinomycetes]

Thu, 24 Mar 2016 23:00:00 +0100

Accurate identification of drug-resistant Mycobacterium tuberculosis is imperative for effective treatment and subsequent reduction in disease transmission. Line probe assays rapidly detect mutations associated with resistance and wild-type sequences associated with susceptibility. Examination of molecular-level performance is necessary for improved assay result interpretation and for continued diagnostic development. Using data collected from a large, multisite diagnostic study, probe hybridization results from line probe assays, MTBDRplus and MTBDRsl, were compared to those of sequencing, and the diagnostic performance of each individual mutation and wild-type probe was assessed. Line probe assay results classified as resistant due to the absence of wild-type probe hybridization were com...



Depletion of Human DNA in Spiked Clinical Specimens for Improvement of Sensitivity of Pathogen Detection by Next-Generation Sequencing [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

In this study, we aimed to develop a specimen-processing protocol to remove human DNA and enrich specimens for bacterial and viral DNA for shotgun metagenomic sequencing. Cerebrospinal fluid (CSF) and nasopharyngeal aspirate (NPA) specimens, spiked with control bacterial and viral pathogens, were processed using either a commercially available kit (MolYsis) or various detergents followed by DNase prior to the extraction of DNA. Relative quantities of human DNA and pathogen DNA were determined by real-time PCR. The MolYsis kit did not improve the pathogen-to-human DNA ratio, but significant reductions (>95%; P < 0.001) in human DNA with minimal effect on pathogen DNA were achieved in samples that were treated with 0.025% saponin, a nonionic surfactant. Specimen preprocessing significa...



Evaluation of a New Device for Simplifying and Standardizing Stool Sample Preparation for Viral Molecular Testing with Limited Hands-On Time [Virology]

Thu, 24 Mar 2016 23:00:00 +0100

Sensitive molecular assays have greatly improved the diagnosis of viral gastroenteritis. However, the proper preparation of stool samples for clinical testing remains an issue. bioMérieux has developed a stool preprocessing device (SPD) that includes a spoon for calibrated sampling and a vial containing buffer, glass beads, and two filters. The resulting stool filtrate is used for nucleic acid extraction. The purpose of this study was to evaluate the performance of the SPD for the quantification of human adenovirus (HAdV) DNA in stool samples collected from hematopoietic stem cell transplant (HSCT) recipients. HAdV DNA was quantified with the Adenovirus R-gene kit. The suitability of the device to reproducibly quantify HAdV DNA in stools using different extraction platforms (easyMAG...



Automated Broad-Range Molecular Detection of Bacteria in Clinical Samples [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Molecular detection methods, such as quantitative PCR (qPCR), have found their way into clinical microbiology laboratories for the detection of an array of pathogens. Most routinely used methods, however, are directed at specific species. Thus, anything that is not explicitly searched for will be missed. This greatly limits the flexibility and universal application of these techniques. We investigated the application of a rapid universal bacterial molecular identification method, IS-pro, to routine patient samples received in a clinical microbiology laboratory. IS-pro is a eubacterial technique based on the detection and categorization of 16S-23S rRNA gene interspace regions with lengths that are specific for each microbial species. As this is an open technique, clinicians do not need to d...



Pneumococcal Colonization Rates in Patients Admitted to a United Kingdom Hospital with Lower Respiratory Tract Infection: a Prospective Case-Control Study [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Current diagnostic tests are ineffective for identifying the etiological pathogen in hospitalized adults with lower respiratory tract infections (LRTIs). The association of pneumococcal colonization with disease has been suggested as a means to increase the diagnostic precision. We compared the pneumococcal colonization rates and the densities of nasal pneumococcal colonization by (i) classical culture and (ii) quantitative real-time PCR (qPCR) targeting lytA in patients with LRTIs admitted to a hospital in the United Kingdom and control patients. A total of 826 patients were screened for inclusion in this prospective case-control study. Of these, 38 patients were recruited, 19 with confirmed LRTIs and 19 controls with other diagnoses. Nasal wash (NW) samples were collected at the time of ...

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Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Aspergillus fumigatus [Mycology]

Thu, 24 Mar 2016 23:00:00 +0100

In this study, the new member of the fungal annexin family, annexin C4, was chosen as the target to design a loop-mediated isothermal amplification (LAMP) assay for the rapid, specific, and sensitive detection of A. fumigatus. The evaluation of the specificity of the LAMP assay that was developed showed that no false-positive results were observed for the 22 non-A. fumigatus strains, including 5 species of the Aspergillus genus. Its detection limit was approximately 10 copies per reaction in reference plasmids, with higher sensitivity than that of real-time quantitative PCR (qPCR) at 102 copies for the same target. Clinical samples from a total of 69 patients with probable IA (n = 14) and possible IA (n = 55) were subjected to the LAMP assay, and positive results were found for the 14 pati...



Cumulative Antimicrobial Susceptibility Data from Intensive Care Units at One Institution: Should Data Be Combined? [Epidemiology]

Thu, 24 Mar 2016 23:00:00 +0100

In this study, susceptibility data for two different ICUs at a tertiary care hospital in Toronto, Canada, were used to create a traditional CSTD report, which combined data from different ICUs, and a rolling-average CSTD report, which pooled 2 years of data for each ICU separately. For simplicity, data for only the most common Gram-negative organisms (Escherichia coli, Pseudomonas aeruginosa) and the most relevant antibiotics (ciprofloxacin, piperacillin-tazobactam) were examined. With the rolling-average method, significant differences in susceptibility were seen between the ICUs in 50% of the organism-antimicrobial combinations. Furthermore, the 3% median year-over-year difference in susceptibilities seen for the 16 organism-antibiotic combinations by using the traditional method was low...



Comparison of Nonculture Blood-Based Tests for Diagnosing Invasive Aspergillosis in an Animal Model [Mycology]

Thu, 24 Mar 2016 23:00:00 +0100

The European Aspergillus PCR Initiative (EAPCRI) has provided recommendations for the PCR testing of whole blood (WB) and serum/plasma. It is important to test these recommended protocols on nonsimulated "in vivo" specimens before full clinical evaluation. The testing of an animal model of invasive aspergillosis (IA) overcomes the low incidence of disease and provides experimental design and control that is not possible in the clinical setting. Inadequate performance of the recommended protocols at this stage would require reassessment of methods before clinical trials are performed and utility assessed. The manuscript describes the performance of EAPCRI protocols in an animal model of invasive aspergillosis. Blood samples taken from a guinea pig model of IA were used for WB and serum PCR....



Pneumococcus with the "6E" cps Locus Produces Serotype 6B Capsular Polysaccharide [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Genetic studies of serogroup 6 isolates of Streptococcus pneumoniae identified putative serotype 6E. Although its capsular polysaccharide structure has not been elucidated, putative serotype 6E is described in an increasing number of studies as a potentially new serotype. We show here that SPEC6B, which is widely used as a target strain for serotype 6B opsonophagocytosis assays, has the genetic features of the putative serotype 6E but produces capsular polysaccharide identical to 6B capsular polysaccharide as determined by one-dimensional (1D) and 2D nuclear magnetic resonance (NMR). Thus, putative serotype 6E is a mere genetic variant of serotype 6B. Also, SPEC6B is appropriate as a target strain for serotype 6B opsonophagocytosis assays. This example illustrates the difficulties of assig...



Development and Validation of an Improved PCR Method Using the 23S-5S Intergenic Spacer for Detection of Rickettsiae in Dermacentor variabilis Ticks and Tissue Samples from Humans and Laboratory Animals [Clinical Veterinary Microbiology]

Thu, 24 Mar 2016 23:00:00 +0100

A novel nested PCR assay was developed to detect Rickettsia spp. in ticks and tissue samples from humans and laboratory animals. Primers were designed for the nested run to amplify a variable region of the 23S-5S intergenic spacer (IGS) of Rickettsia spp. The newly designed primers were evaluated using genomic DNA from 11 Rickettsia species belonging to the spotted fever, typhus, and ancestral groups and, in parallel, compared to other Rickettsia-specific PCR targets (ompA, gltA, and the 17-kDa protein gene). The new 23S-5S IGS nested PCR assay amplified all 11 Rickettsia spp., but the assays employing other PCR targets did not. The novel nested assay was sensitive enough to detect one copy of a cloned 23S-5S IGS fragment from "Candidatus Rickettsia amblyommii." Subsequently, the detection...

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Propionibacterium acnes: Disease-Causing Agent or Common Contaminant? Detection in Diverse Patient Samples by Next-Generation Sequencing [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Propionibacterium acnes is the most abundant bacterium on human skin, particularly in sebaceous areas. P. acnes is suggested to be an opportunistic pathogen involved in the development of diverse medical conditions but is also a proven contaminant of human clinical samples and surgical wounds. Its significance as a pathogen is consequently a matter of debate. In the present study, we investigated the presence of P. acnes DNA in 250 next-generation sequencing data sets generated from 180 samples of 20 different sample types, mostly of cancerous origin. The samples were subjected to either microbial enrichment, involving nuclease treatment to reduce the amount of host nucleic acids, or shotgun sequencing. We detected high proportions of P. acnes DNA in enriched samples, particularly skin tis...



Direct Identification of Urinary Tract Pathogens from Urine Samples, Combining Urine Screening Methods and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Early diagnosis of urinary tract infections (UTIs) is essential to avoid inadequate or unnecessary empirical antibiotic therapy. Microbiological confirmation takes 24 to 48 h. The use of screening methods, such as cytometry and automated microscopic analysis of urine sediment, allows the rapid prediction of negative samples. In addition, matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is a widely established technique in clinical microbiology laboratories used to identify microorganisms. We evaluated the ability of MALDI-TOF MS to identify microorganisms from direct urine samples and the predictive value of automated analyzers for the identification of microorganisms in urine by MALDI-TOF MS. A total of 451 urine samples from patients with ...



Reassessment of the Role of Rapid Antigen Detection Tests in Diagnosis of Invasive Group A Streptococcal Infections [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Rapid antigen detection tests (RADTs) for group A streptococci (GAS) are widely used for diagnosing acute pharyngitis, which has led to a considerable reduction in antibiotic prescriptions over the past decade. Beyond this intended use, their reassessment on invasive samples may be relevant in the management of life-threatening GAS infections. To this end, we evaluated the performances of three RADTs, culture, GAS PCR, and 16S rRNA gene PCR assays, and compared them with a composite gold standard (GAS-PCR assay and/or culture) for the diagnosis of severe GAS infection. A total of 192 specimens from deep-tissue (mostly normally sterile) sites enriched for 75 GAS-positive samples were enrolled in the study. The three evaluated RADTs showed sensitivities ranging from 88.0% to 94.7% versus 98....



Unbiased Detection of Respiratory Viruses by Use of RNA Sequencing-Based Metagenomics: a Systematic Comparison to a Commercial PCR Panel [Virology]

Thu, 24 Mar 2016 23:00:00 +0100

Current infectious disease molecular tests are largely pathogen specific, requiring test selection based on the patient's symptoms. For many syndromes caused by a large number of viral, bacterial, or fungal pathogens, such as respiratory tract infections, this necessitates large panels of tests and has limited yield. In contrast, next-generation sequencing-based metagenomics can be used for unbiased detection of any expected or unexpected pathogen. However, barriers for its diagnostic implementation include incomplete understanding of analytical performance and complexity of sequence data analysis. We compared detection of known respiratory virus-positive (n = 42) and unselected (n = 67) pediatric nasopharyngeal swabs using an RNA sequencing (RNA-seq)-based metagenomics approach and Taxono...



Detecting Staphylococcus aureus Virulence and Resistance Genes: a Comparison of Whole-Genome Sequencing and DNA Microarray Technology [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

In conclusion, WGS may substitute array-based methods due to its universal methodology, open and expandable nature, and rapid parallel analysis capacity for different characteristics in once-generated sequences. (Source: Journal of Clinical Microbiology)

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Development and Validation of a Highly Accurate Quantitative Real-Time PCR Assay for Diagnosis of Bacterial Vaginosis [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

Bacterial vaginosis (BV) is the most common gynecological infection in the United States. Diagnosis based on Amsel's criteria can be challenging and can be aided by laboratory-based testing. A standard method for diagnosis in research studies is enumeration of bacterial morphotypes of a Gram-stained vaginal smear (i.e., Nugent scoring). However, this technique is subjective, requires specialized training, and is not widely available. Therefore, a highly accurate molecular assay for the diagnosis of BV would be of great utility. We analyzed 385 vaginal specimens collected prospectively from subjects who were evaluated for BV by clinical signs and Nugent scoring. We analyzed quantitative real-time PCR (qPCR) assays on DNA extracted from these specimens to quantify nine organisms associated w...



Accurate Serodetection of Asymptomatic Leishmania donovani Infection by Use of Defined Antigens [Parasitology]

Thu, 24 Mar 2016 23:00:00 +0100

Infection with Leishmania donovani is typically asymptomatic, but a significant number of individuals may progress to visceral leishmaniasis (VL), a deadly disease that threatens 200 million people in areas where it is endemic. While diagnosis of acute VL has been simplified by the use of cost-effective confirmatory serological tests, similar standardized tools are not widely available for detecting asymptomatic infection, which can be 4 to 20 times more prevalent than active disease. A simple and accurate serological test that is capable of detecting asymptomatic L. donovani infection will be useful for surveillance programs targeting VL control and elimination. To address this unmet need, we evaluated recombinant antigens for their ability to detect serum antibodies in 104 asymptomatic L...



U.S. Ebola Treatment Center Clinical Laboratory Support [Virology]

Thu, 24 Mar 2016 23:00:00 +0100

This study determined the clinical laboratory capabilities of these ETCs. ETCs were electronically surveyed on clinical laboratory characteristics. Survey responses were returned from 47 ETCs (85%). Forty-one (87%) of the ETCs planned to provide some laboratory support (e.g., point-of-care [POC] testing) within the room of the isolated patient. Forty-four (94%) ETCs indicated that their hospital would also provide clinical laboratory support for patient care. Twenty-two (50%) of these ETC clinical laboratories had biosafety level 3 (BSL-3) containment. Of all respondents, 34 (72%) were supported by their jurisdictional public health laboratory (PHL), all of which had available BSL-3 laboratories. Overall, 40 of 44 (91%) ETCs reported BSL-3 laboratory support via their clinical laboratory a...



Evaluation of Performance Characteristics of the Aptima HIV-1 Quant Dx Assay for Detection and Quantitation of Human Immunodeficiency Virus Type 1 in Plasma and Cervicovaginal Lavage Samples [Virology]

Thu, 24 Mar 2016 23:00:00 +0100

The objective of this study was to evaluate performance characteristics and relative workflow of the Aptima HIV-1 Quant Dx assay in comparison with the Abbott RealTime HIV-1 assay using plasma and cervicovaginal lavage (CVL) specimens. Assay performance was evaluated by using an AcroMetrix HIV-1 panel, AcroMetrix positive controls, Qnostics and SeraCare HIV-1 evaluation panels, 208 clinical plasma samples, and 205 matched CVL specimens on the Panther and m2000 platforms. The Aptima assay demonstrated good linearity over the quantification range tested (2 to 5 log10 copies/ml), and there was strong linear correlation between the assays (R2 = 0.99), with a comparable coefficient of variance of <5.5%. For the plasma samples, Deming regression analyses and Bland-Altman plots showed excellen...



Evaluation of Polymorphic Locus Sequence Typing for Candida glabrata Epidemiology [Epidemiology]

Thu, 24 Mar 2016 23:00:00 +0100

The opportunistic yeast Candida glabrata is increasingly refractory to antifungal treatment or prophylaxis and relatedly is increasingly implicated in health care-associated infections. To elucidate the epidemiology of these infections, strain typing is required. Sequence-based typing provides multiple advantages over length-based methods, such as pulsed-field gel electrophoresis (PFGE); however, conventional multilocus sequence typing (targeting 6 conserved loci) and whole-genome sequencing are impractical for routine use. A commercial sequence-based typing service for C. glabrata that targets polymorphic tandem repeat-containing loci has recently been developed. These CgMT-J and CgMT-M services were evaluated with 56 epidemiologically unrelated isolates, 4 to 7 fluconazole-susceptible or...

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Analytical and Clinical Evaluation of the Epistem Genedrive Assay for Detection of Mycobacterium tuberculosis [Mycobacteriology and Aerobic Actinomycetes]

Thu, 24 Mar 2016 23:00:00 +0100

The Epistem Genedrive assay rapidly detects the Mycobacterium tuberculosis complex from sputum and is currently available for clinical use. However, the analytical and clinical performance of this test has not been fully evaluated. The analytical limit of detection (LOD) of the Genedrive PCR amplification was tested with genomic DNA; the performance of the complete (sample processing plus amplification) system was tested by spiking M. tuberculosis mc26030 cells into distilled water and M. tuberculosis-negative sputum. Specificity was tested using common respiratory pathogens and nontuberculosis mycobacteria. A clinical evaluation enrolled adults with suspected pulmonary tuberculosis, obtained three sputum samples from each participant, and compared the accuracy of the Genedrive to that of ...



Development of a Highly Specific IgM Enzyme-Linked Immunosorbent Assay for Bartonella henselae Using Refined N-Lauroyl-Sarcosine-Insoluble Proteins for Serodiagnosis of Cat Scratch Disease [Immunoassays]

Thu, 24 Mar 2016 23:00:00 +0100

The conventional anti-Bartonella henselae IgM enzyme-linked immunosorbent assay (IgM-ELISA) methods for diagnosing cat scratch disease (CSD) remain poor in both sensitivity and specificity. We sought to develop an IgM-ELISA with improved accuracy in the serodiagnosis of CSD by exploring the antigens that are most suitable for an ELISA. We prepared 5 different protein antigens: antigen I (sonicated B. henselae whole-cell antigen), antigen II (N-lauroyl-sarcosine-insoluble antigen), antigen III (processed sarcosine-soluble antigen), and antigen IV and antigen V (sarcosine-insoluble and sarcosine-soluble antigens refined by DEAE-Sepharose Fast Flow ion-exchange chromatography). The IgM antibodies in the sera of 47 patients with clinically suspected CSD (24 definite, 23 suspected) and of 85 he...



Multiplex Antibody Detection for Noninvasive Genus-Level Diagnosis of Prosthetic Joint Infection [Bacteriology]

Thu, 24 Mar 2016 23:00:00 +0100

We developed and evaluated a multiplex antibody detection-based immunoassay for the diagnosis of prosthetic joint infections (PJIs). Sixteen protein antigens from three Staphylococcus species (Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus lugdunensis) (8 antigens), Streptococcus agalactiae (4 antigens), and Propionibacterium acnes (4 antigens) were selected by comparative immunoproteomics using serum samples from PJI cases versus controls. A bead-based multiplex immunoassay that measured serum IgG against purified, recombinant forms of each of the 16 antigens was developed. We conducted a prospective study to evaluate the performance of the assay. A PJI was defined by the presence of a sinus tract and/or positive intraoperative sample cultures (at least one sample y...