کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
9263375 1216034 2005 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Rapid detection of mecA and nuc genes in staphylococci by real-time multiplex polymerase chain reaction
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی میکروبیولوژی و بیوتکنولوژی کاربردی
پیش نمایش صفحه اول مقاله
Rapid detection of mecA and nuc genes in staphylococci by real-time multiplex polymerase chain reaction
چکیده انگلیسی
A multiplex real-time polymerase chain reaction (RT-PCR) targeting the mecA and nuc genes was developed for the detection of methicillin resistance and identification of Staphylococcus aureus. Novel mecA and nuc primers and fluorescence resonance energy transfer hybridization probes specific for the mecA and nuc genes were evaluated. The assay was performed using the LightCycler system (Roche Molecular Biochemicals, Mannheim, Germany) and evaluated against the traditional gel-based multiplex PCR (PCR-gel) method currently used at Royal Perth Hospital. Clinical isolates (n = 222) and isolates from a culture collection library (n = 206) were tested by both assays in parallel. The RT-PCR assay was 100% sensitive and specific for the detection of methicillin resistance and for the identification of S. aureus when compared with the PCR-gel assay. Results from the RT-PCR assay showed 5 isolates with lower efficiency fluorescence curves for the nuc gene PCR fragment. DNA sequencing showed mutations within the region of the probe-binding sites compared with the reference strain. The results of the RT-PCR assay were available within 2 h. This rapid mecA/nuc RT-PCR assay is a suitable and practical tool for the routine detection of methicillin resistance and identification of S. aureus, which can be easily incorporated into the diagnostic molecular microbiology laboratory work flow.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Diagnostic Microbiology and Infectious Disease - Volume 51, Issue 1, January 2005, Pages 13-17
نویسندگان
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