ArticleJournal of clinical microbiology2005
Species-level identification of staphylococcal isolates by real-time PCR and melt curve analysis.
Article in Journal of clinical microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 70 citations in OpenAlex.
- Distinct T cell signatures are associated with Staphylococcus aureus skin infection in pediatric atopic dermatitis.JCI insight · 2024Article
- DNA sequencing, genomes and genetic markers of microbes on fruits and vegetables.Microbial biotechnology · 2021Review
- Usefulness of Multiplex Real-Time PCR for Simultaneous Pathogen Detection and Resistance Profiling of Staphylococcal Bacteremia.BioMed research international · 2016Article
- CodY-mediated regulation of the Staphylococcus aureus Agr system integrates nutritional and population density signals.Journal of bacteriology · 2014Article
- Rapid and accurate identification of human-associated staphylococci by use of multiplex PCR.Journal of clinical microbiology · 2011Article
- Real-time detection of blaKPC in clinical samples and surveillance specimens.Journal of clinical microbiology · 2011Article
- Fulminant staphylococcus lugdunensis septicaemia following a pelvic varicella-zoster virus infection in an immune-deficient patient: a case report.European journal of medical research · 2010Article
- fbl gene as a species-specific target for Staphylococcus lugdunensis identification.Journal of clinical laboratory analysis · 2010Article
- Effect of cinnamon oil on icaA expression and biofilm formation by Staphylococcus epidermidis.Applied and environmental microbiology · 2009Article
- Comparison of real-time PCR and conventional biochemical methods for identification of Staphylococcus lugdunensis.Journal of clinical microbiology · 2009Article
- Composition of bacterial communities associated with natural and laboratory populations of Asobara tabida infected with Wolbachia.Applied and environmental microbiology · 2009Article
- Identification of Staphylococcus species directly from positive blood culture broth by use of molecular and conventional methods.Journal of clinical microbiology · 2009Article
- Codon-optimized fluorescent proteins designed for expression in low-GC gram-positive bacteria.Applied and environmental microbiology · 2009Article
- recA mediated spontaneous deletions of the icaADBC operon of clinical Staphylococcus epidermidis isolates: a new mechanism of phenotypic variations.Antonie van Leeuwenhoek · 2008Article
- High-throughput avian molecular sexing by SYBR green-based real-time PCR combined with melting curve analysis.BMC biotechnology · 2008Article
- From clinical microbiology to infection pathogenesis: how daring to be different works for Staphylococcus lugdunensis.Clinical microbiology reviews · 2008Review
- Rapid identification of pathogenic yeast isolates by real-time PCR and two-dimensional melting-point analysis.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2007Article
- A detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria.Journal of microbiological methods · 2007Article
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A real-time PCR assay was developed to identify common staphylococcal species. A single set of consensus primers was designed to amplify a portion of the 16S rRNA gene, and a pair of fluorescence resonance energy transfer probes was used to identify species based on the unique melt properties of the probes resulting from sequence variations in the amplicons from each species. Nine common staphylococcal strains (S. aureus, S. capitis, S. epidermidis, S. haemolyticus, S. hominis, S. lugdunensis, S. schleiferi, S. simulans, and S. warneri) were used for assay development. The species-specific melting profiles were validated by correctly identifying 36 of 37 coagulase-negative staphylococcal (CoNS) isolates identified by ribotyping. In a study of clinical isolates, the PCR/melt curve approach correctly identified 56/56 S. aureus isolates identified by coagulase/protein A latex agglutination. Fifty-four CoNS clinical isolates characterized using the API Staph assay were studied, with the PCR/melt curve approach yielding matching identifications for 32/54 (59%). The API Staph assay was unable to identify 18 CoNS isolates, and differing results were obtained for 4 isolates. Sequencing of the 22 discrepant or unidentified CoNS samples revealed that the PCR/melt curve results were correct for all but one isolate. Thus, PCR/melt curve analysis achieved a nearly 100% accuracy and performed better than biochemical testing. Performance of the PCR/melt curve approach requires less than 2 h after colony selection. This method thus provides a rapid and accurate approach to the identification of staphylococcal species in the clinical laboratory.
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