ArticleJournal of clinical microbiology2000
A 5' nuclease PCR (TaqMan) high-throughput assay for detection of the mecA gene in staphylococci.
Article in Journal of clinical microbiology, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- A pseudo-outbreak of MRSA due to laboratory contamination related to MRSA carriage of a laboratory staff member.Antimicrobial resistance and infection control · 2023Article
- Right-sided endocarditis caused by polyclonal Staphylococcus aureus infection.European journal of medical research · 2021Article
- Mupirocin Resistance in Isolates of Staphylococcus spp. from Nasal Swabs in a Tertiary Hospital in France.Journal of clinical microbiology · 2015Article
- Evaluation of the BD GeneOhm methicillin-resistant Staphylococcus aureus (MRSA) assay as a method for detection of MRSA isolates, using a large collection of European and North African isolates.Journal of clinical microbiology · 2014Article
- Ward-specific rates of nasal cocolonization with methicillin-susceptible and -resistant Staphylococcus spp. and potential impact on molecular methicillin-resistant Staphylococcus aureus screening tests.Journal of clinical microbiology · 2013Article
- Prevalence and characterization of Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus, isolated from bulk tank milk from Minnesota dairy farms.Journal of clinical microbiology · 2012Article
- Multiplex real-time PCR assay for detection of methicillin-resistant Staphylococcus aureus and associated toxin genes.Journal of clinical microbiology · 2011Article
- Predicting the sensitivity and specificity of published real-time PCR assays.Annals of clinical microbiology and antimicrobials · 2008Article
- Characterization of Staphylococcus aureus isolates from nasal cultures collected from individuals in the United States in 2001 to 2004.Journal of clinical microbiology · 2008Article
- Accuracy and potential usefulness of triplex real-time PCR for improving antibiotic treatment of patients with blood cultures showing clustered gram-positive cocci on direct smears.Journal of clinical microbiology · 2008Article
- Real-time PCR in clinical microbiology: applications for routine laboratory testing.Clinical microbiology reviews · 2006Review
- Molecular evolution of methicillin-resistant Staphylococcus aureus in the metropolitan area of Cologne, Germany, from 1984 to 1998.Journal of clinical microbiology · 2005Article
- Detection of methicillin-resistant Staphylococcus aureus directly from nasal swab specimens by a real-time PCR assay.Journal of clinical microbiology · 2004Article
- Origins of community strains of methicillin-resistant Staphylococcus aureus.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2004Article
- Development of a real-time fluorescence PCR assay for rapid detection of the diphtheria toxin gene.Journal of clinical microbiology · 2002Article
- Modeling of 5' nuclease real-time responses for optimization of a high-throughput enrichment PCR procedure for Salmonella enterica.Journal of clinical microbiology · 2002Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In an effort to find a rapid, efficient, and reliable method of screening large numbers of bacterial isolates for specific antimicrobial resistance genes, we compared conventional PCR results to the results generated using the TaqMan 5' nuclease PCR kit in conjunction with an ABI Prism 7700 Sequence Detector for detecting the mecA gene in various species of staphylococci. DNA was extracted using two techniques. The first used a high-salt extraction method suitable for conventional PCR but resulted in a 7.2% rate of PCR inhibition with the TaqMan technique. PCR inhibition could be overcome by diluting samples 1:5 prior to testing. The second method used the Qiagen QIAamp Tissue Kit; no instances of PCR inhibition were encountered with this method. A total of 197 (96%) of the 206 samples with no inhibition showed agreement between the two methods. Eight of the nine disagreements were likely the result of low-level DNA cross contamination caused by frequent specimen handling. Target DNA in all eight of these samples was first detected in the initial tests only after >30 PCR cycles, and all were negative upon repeat testing even after 40 PCR cycles using freshly extracted DNA. Among those positive samples in agreement, target DNA was invariably detected before 30 PCR cycles. The TaqMan assay eliminated the need to load, run, stain, and read agarose gels and provided the advantage of instant detection of PCR product by laser-activated fluorescence. Thus, final results were obtained 2 h after PCR was initiated, as opposed to a requirement of 2 days to examine 96 samples by agarose gel electrophoresis.
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