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Journal of Clinical Microbiology

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https://www.readbyqxmd.com/read/28814586/molecular-diagnosis-of-invasive-aspergillosis-and-detection-of-azole-resistance-by-a-newly-commercialized-pcr-kit
#1
Eric Dannaoui, Frédéric Gabriel, Manuel Gaboyard, Gaëlle Lagardere, Lucile Audebert, Gilles Quesne, Sandrine Godichaud, Paul E Verweij, Isabelle Accoceberry, Marie-Elisabeth Bougnoux
Aspergillus fumigatus is the main species responsible for aspergillosis in humans. The diagnosis of aspergillosis remains difficult, and the rapid emergence of azole resistance in A. fumigatus, is worrisome. The aim of this study was to validate the new MycoGENIE® A. fumigatus real-time PCR kit and to evaluate its performance in clinical samples for the detection of A. fumigatus and its azole resistance. This multiplex assay detects DNA from the A. fumigatus species complex by targeting the 28S-rRNA multicopy gene and specific TR34 and L98H mutations in the cyp51A single copy gene of A...
August 16, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28814585/a-multiplex-rt-pcr-assay-for-the-simultaneous-identification-of-neisseria-gonorrhoeae-and-its-ciprofloxacin-susceptibility-status
#2
Sumudu R Perera, Nurul H Khan, Irene Martin, Ali Taheri, Rajinder P Parti, Paul N Levett, Greg B Horsman, Anthony Kusalik, Jo-Anne R Dillon
A real time PCR (RT-PCR) assay was designed for the simultaneous identification of N. gonorrhoeae and its ciprofloxacin susceptibility status. A SYBR-Green based multiplex RT-PCR format was used and comprised two different forward primers and a common reverse primer to detect single nucleotide polymorphisms (SNPs) in gyrA of N. gonorrhoeae The primer pairs were evaluated for their sensitivity and specificity with genomic DNA from 254 N. gonorrhoeae isolates (82 were ciprofloxacin susceptible and 172 were ciprofloxacin resistant) and 23 non-N...
August 16, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794183/french-prospective-clinical-evaluation-of-the-aptima%C3%A2-mycoplasma-genitalium-assay-ce-ivd-and-macrolide-resistance-detection-using-three-distinct-assays
#3
Chloé Le Roy, Sabine Pereyre, Nadège Hénin, Cécile Bébéar
The aim of this study was to evaluate the clinical performance of the Aptima® Mycoplasma genitalium CE-IVD (MG-TMA) assay for the detection of Mycoplasma genitalium in clinical male and female samples in comparison with the in-house real-time PCR (in-house PCR) assay routinely used in our laboratory. A total of 1,431 clinical specimens obtained from 1,235 patients were prospectively collected at the Bacteriology Department of Bordeaux University Hospital (France). Additional research-use-only Aptima® M. genitalium TMA assays, Alt1-TMA and Alt2-TMA, were performed on discordant specimens to determine the M...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794182/membrane-filtration-is-suitable-to-reliably-eliminate-mycobacterium-tuberculosis-from-saliva-for-therapeutic-drug-monitoring
#4
Simone H J van den Elsen, Tridia van der Laan, Onno W Akkerman, Adri G M van der Zanden, Jan-Willem C Alffenaar, Dick van Soolingen
Tuberculosis (TB) remains an infectious disease of a worldwide concern.….
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794181/a-neutralization-assay-for-zika-and-dengue-viruses-using-a-real-time-pcr-based-endpoint-assessment
#5
Heather L Wilson, Thomas Tran, Julian Druce, Myrielle Dupont-Rouzeyrol, Michael Catton
The global spread and infective complications of Zika virus (ZKV) and Dengue virus (DENV) have made them flaviviruses of public health concern. Serological diagnosis can be challenging due to antibody cross-reactivity, particularly in secondary flavivirus infections or when there is a history of flavivirus vaccination. The virus neutralization assay is considered to be the most specific assay for measurement of anti-flavivirus antibodies. This study describes an assay where neutralization endpoint is measured by real-time PCR, providing results within 72 hours...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794180/determination-of-disk-diffusion-and-mic-quality-control-guidelines-for-high-dose-cefepime-tazobactam-wck-4282-a-novel-antibacterial-combination-consisting-of-a-%C3%AE-lactamase-inhibitor-and-a-fourth-generation-cephalosporin
#6
Stefan Riedel, Michael D Huband, Helio S Sader, Robert K Flamm, Ronald N Jones
High-dose cefepime-tazobactam (1:1; WCK 4282), a novel antibacterial combination consisting of the β-lactamase inhibitor tazobactam and a fourth-generation cephalosporin, is under clinical development for the therapy of serious gram-negative infections. A quality control (QC) study was performed to establish disk diffusion and MIC ranges for cefepime-tazobactam for multiple QC reference strains. The cefepime-tazobactam QC ranges for MIC (tazobactam at fixed 8 μg/mL) and disk diffusion (30/20-μg disk) test methods were approved by the CLSI Subcommittee on Antimicrobial Susceptibility Testing in January 2015 and January 2016...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794179/evaluation-of-the-copan-fecalswab%C3%A2-as-specimen-type-for-use-in-xpert%C3%A2-c-difficile-assay
#7
Michael J Mashock, Matthew L Faron, Blake W Buchan, Nathan A Ledeboer
Liquid based microbiology (LBM) devices incorporating flocked swabs and preservation medium eases transport of specimens and improves specimen yield compared to traditional fiber wound swabs; however, the performance of LBM collection devices has not been evaluated in many molecular assays. It is unclear how the differences in matrix and specimen loading with an LBM device will affect test performance compared to traditional collection devices. The purpose of this study was to evaluate the performance of specimens collected in FecalSwab™ transport medium (Copan Diagnostics, Murrieta CA) compared to unpreserved stool using the Cepheid Xpert C...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794178/potential-of-high-affinity-slow-off-rate-modified-aptamer-somamer-reagents-for-mycobacterium-tuberculosis-proteins-as-tools-for-infection-models-and-diagnostic-applications
#8
Theresa Russell, Louis S Green, Taylor Rice, Nicole A Kruh-Garcia, Karen Dobos, Mary A DeGroote, Thomas Hraha, David G Sterling, Nebojsa Janjic, Urs A Ochsner
Direct pathogen detection in blood to diagnose active tuberculosis (TB) has been difficult, due to low levels of circulating antigens or due to the lack of specific, high-affinity binding reagents and reliable assays with adequate sensitivity. We sought to determine whether slow off-rate modified aptamer (SOMAmer) reagents with sub-nanomolar affinity for M. tuberculosis (Mtb) proteins (antigen 85A, 85B, 85C, GroES, GroEL2, DnaK, CFP10, KAD, CFP2, RplL and Tpx) could be useful to diagnose tuberculosis. When incorporated into the multiplexed, array-based proteomic SOMAscan assay, limits of detection reached the sub-picomolar range in 40% serum...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794177/discovery-and-validation-of-a-six-marker-serum-protein-signature-for-the-diagnosis-of-active-pulmonary-tuberculosis
#9
Mary A De Groote, David G Sterling, Thomas Hraha, Theresa Russell, Louis S Green, Kirsten Wall, Stephan Kraemer, Rachel Ostroff, Nebojsa Janjic, Urs A Ochsner
New non-sputum biomarker tests for active tuberculosis (TB) diagnostics are of highest priority for global TB control. We performed in-depth proteomic analysis using the 4000-plex SOMAscan assay on 1470 serum samples from seven TB-endemic countries. All samples were from patients with symptoms and signs suggestive of active pulmonary TB that were systematically confirmed or ruled-out for TB by culture and clinical follow-up. HIV-coinfection was present in 34%and 22% were sputum smear-negative. Serum protein biomarkers were identified by stability selection using L1-regularized logistic regression and by Kolmogorov-Smirnov (KS) statistics...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794176/molecular-tests-that-target-the-rtx-locus-do-not-distinguish-between-kingella-kingae-and-the-recently-described-kingella-negevensis-species
#10
Nawal El Houmami, Janek Bzdrenga, Guillaume André Durand, Philippe Minodier, Hervé Seligmann, Elsa Prudent, Sofiane Bakour, Stéphane Bonacorsi, Didier Raoult, Pablo Yagupsky, Pierre-Edouard Fournier
Kingella kingae is an important invasive pathogen in early childhood. The organism elaborates a RTX toxin presumably restricted to this species. Consequently, real-time PCR (qPCR) assays targeting the RTX locus have been developed in recent years and are gaining increasing use for the molecular diagnosis of K. kingae infections. However, this study shows that Kingella negevensis, a Kingella species newly identified in young children, harbors an identical Kingella RTX locus, raising the question whether K. negevensis could be misdiagnosed as K...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28794175/a-comparative-genomics-study-of-staphylococcus-epidermidis-from-orthopedic-device-related-infections-correlated-with-patient-outcome
#11
Virginia Post, Llinos G Harris, Mario Morgenstern, Leonardos Mageiros, Matthew D Hitchings, Guillaume Méric, Ben Pascoe, Samuel K Sheppard, R Geoff Richards, T Fintan Moriarty
Staphylococcus epidermidis has emerged as an important opportunistic pathogen causing orthopedic device-related infections (ODRIs). This study investigated the association of genome variation and phenotypic features of the infecting S. epidermidis isolate with the clinical outcome of the infected patient. S. epidermidis isolates were collected from 104 patients with ODRI. Their clinical outcome was evaluated, after an average of 26 months, as either "cured" or "not cured". The isolates were tested for antibiotic susceptibility and biofilm formation...
August 9, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28768732/evaluation-of-a-new-commercial-microarray-platform-for-the-simultaneous-detection-of-%C3%AE-lactamase-and-mcr-1-2-genes-in-enterobacteriaceae
#12
Odette J Bernasconi, Luigi Principe, Regula Tinguely, Aneta Karczmarek, Vincent Perreten, Francesco Luzzaro, Andrea Endimiani
The worldwide prevalence of infections due to Gram-negative bacteria co-resistant to extended-spectrum cephalosporins and carbapenems is worrisome.….
August 2, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28768731/improved-serodiagnostic-performance-for-lyme-disease-by-use-of-two-recombinant-proteins-in-elisa-as-compared-to-standardized-two-tier-testing
#13
Gary L Bradshaw, R Kelley Thueson, Todd J Uriona
The most reliable test method for the serological confirmation of Lyme Disease (LD) is a 2-tiered method recommended by the CDC in 1995. The first-tier test is a low specificity ELISA test and the second-tier tests are higher specificity IgG and IgM Western blots. This study describes the selection of two Borrelia burgdorferi-recombinant proteins and evaluation of their performance in a simple 1-tier test for the serological confirmation of LD. These two proteins were generated from (a) the full length dbpA gene combined with the invariable region 6 of the VlsE gene (dbpA/C6) and (b) the full length OspC gene...
August 2, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28768730/detection-of-plasmodium-spp-infection-by-the-illumigene%C3%A2-malaria-assay-compared-to-reference-microscopy-and-real-time-pcr
#14
Candace Rypien, Barbara Chow, Wilson Chan, Deirdre Church, Dylan R Pillai
Malaria is one of the leading causes of infectious disease in returning travelers from the tropics. Diagnosis of malaria is typically performed by examination of Giemsa-stained thick and thin peripheral blood smears, which is time-consuming, labor-intensive and requires high levels of proficiency. Alternatively, loop-mediated isothermal amplification (LAMP) is a new molecular method, which is rapid, sensitive, and requires less capital equipment and technological training. We conducted a retrospective study comparing two formats of a commercial LAMP assay (Meridian illumigene® Malaria [M] and Malaria Plus [MP]) versus reference microscopy on archived blood specimens (n=140) obtained from unique returning travelers suspected of having malaria...
August 2, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28768729/evaluation-of-a-novel-reporter-virus-neutralization-test-for-the-serological-diagnosis-of-zika-and-dengue-virus-infection
#15
Chao Shan, Daniel A Ortiz, Yujiao Yang, Susan J Wong, Laura D Kramer, Pei-Yong Shi, Michael J Loeffelholz, Ping Ren
The current laboratory diagnosis of Zika virus (ZIKV) infection is primarily through the detection of ZIKV RNA or antibodies against ZIKV proteins. The detection of viral RNA is highly sensitive and specific, but viremia and viruria periods are brief, limiting the utility of ZIKV RNA assays. Instead, most ZIKV infections are diagnosed serologically using the IgM antibody capture enzyme linked immunosorbent assay (MAC-ELISA) for screening followed by a confirmatory plaque reduction neutralization test (PRNT)...
August 2, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28747375/evaluation-of-the-rapid-polymyxin-np-test-for-polymyxin-b-resistance-detection-using-enterobacter-cloacae-and-enterobacter-aerogenes-isolates
#16
Shelby Simar, Diane Sibley, Deborah Ashcraft, George Pankey
Background: Polymyxin resistance is an increasing problem worldwide. Currently, determining susceptibility to polymyxins is problematic and lengthy. Polymyxins diffuse poorly into agar, potentially giving inaccurate disk diffusion and Etest results. A rapid screening test (2 h) for detection of polymyxin-resistance in Enterobacteriaceae, developed by Nordmann and Poirel (Rapid Polymyxin NP Test, 2016), detects glucose metabolization in the presence of polymyxins E and B via pH-induced color change. Sensitivity/specificity were 99...
July 26, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28747374/babesia-microti-from-mice-to-ticks-to-an-increasing-number-of-highly-susceptible-humans
#17
Lars F Westblade, Matthew S Simon, Blaine A Mathison, Laura A Kirkman
Babesia microti, a zoonotic intraerythrocytic parasite, is the primary etiological agent of human babesiosis in the United States (US). Human infection ranges from subclinical illness to severe disease resulting in death, with symptoms related to host immune status. Despite advances in our understanding and management of B. microti, incidence of infection has increased in the US. Therefore, research focused on eradicating disease and optimizing clinical management is essential. Herein, we review this remarkable organism with emphasis on the clinical, diagnostic, and therapeutic aspects of human disease...
July 26, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28747373/investigation-of-respiratory-syncytial-virus-outbreak-on-an-adult-stem-cell-transplant-unit-using-whole-genome-sequencing
#18
Yijun Zhu, Teresa R Zembower, Kristen E Metzger, Zhengdeng Lei, Stefan J Green, Chao Qi
A viral whole genome sequencing strategy, based on PCR amplification followed by Next-generation Sequencing, was used to investigate a nosocomial RSV-B outbreak in a hematology-oncology and stem cell transplant unit. RSV-B genomes from 16 patients and healthcare workers (HCWs) suspected to be involved in the outbreak were compared to RSV-B genomes acquired from outpatients during the same time period but epidemiologically unrelated to the outbreak. Phylogenetic analysis of the whole genome identified a cluster of 11 patients and HCWs with an identical RSV-B strain which were clearly distinct from strains recovered from individuals unrelated to the outbreak...
July 26, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28747372/determination-of-disk-diffusion-and-mic-quality-control-ranges-for-nafithromycin-wck-4873-a-new-lactone-ketolide
#19
Meredith A Hackel, James A Karlowsky, Dana Dressel, Daniel F Sahm
Disk diffusion and MIC quality control (QC) ranges were determined for nafithromycin, a new lactone-ketolide, following completion of a nine-laboratory, Clinical and Laboratory Standards Institute (CLSI) M23-defined Tier 2 study. Five QC strains consistent with the spectrum of activity of nafithromycin were tested: Staphylococcus aureus ATCC 25923 (disk only), S. aureus ATCC 29213 (broth only), Enterococcus faecalis ATCC 29212 (broth only), Streptococcus pneumoniae ATCC 49619 (disk and broth), and Haemophilus influenzae ATCC 49247 (disk and broth)...
July 26, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28747371/designing-hiv-testing-algorithms-based-on-2015-who-guidelines-using-data-from-six-sites-in-sub-saharan-africa
#20
Cara S Kosack, Leslie Shanks, Greet Beelaert, Tumwesigye Benson, Aboubacar Savane, Anne Ng'ang'a, Bita Andre, Jean-Paul B N Zahinda, Katrien Fransen, Anne-Laure Page
Our objective was to evaluate the performance of HIV testing algorithms based on WHO recommendations, using data from specimens collected at six HIV testing and counselling sites in sub-Saharan Africa (Guinea, Conakry; Kitgum and Arua, Uganda; Homa Bay, Kenya; Douala, Cameroun; Baraka, Democratic Republic of Congo). A total of 2780 samples, including 1306 HIV-positive, were included in the analysis. HIV testing algorithms were designed using Determine as a first test. Second and third rapid diagnostic tests (RDT) were selected based on site-specific performance, adhering where possible to the WHO-recommended minimum requirements of sensitivity and specificity of ≥99%...
July 26, 2017: Journal of Clinical Microbiology
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