کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3398663 1222309 2007 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Standard and real-time multiplex PCR methods for detection of trimethoprim resistance dfr genes in large collections of bacteria
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی میکروب شناسی
پیش نمایش صفحه اول مقاله
Standard and real-time multiplex PCR methods for detection of trimethoprim resistance dfr genes in large collections of bacteria
چکیده انگلیسی

ABSTRACTTwo multiplex PCR (mPCR) methods were developed to screen large collections of trimethoprim-resistant Escherichia coli isolates for the most prevalent resistance determinants. Five common integron-carried genes (dfrA1, dfrA5, dfrA7, dfrA12 and dfrA17) were selected as PCR targets. Primers and conditions for standard mPCRs and real-time mPCRs were selected and tested. Two protocols using essentially the same primer pairs were established. The standard mPCR protocol also included an internal control targeting the E. coli 16S rRNA gene. Both protocols proved to be sensitive and specific for detection of the five selected genes. Screening of three different collections of clinical urinary and blood isolates (n = 368) with the two multiplex methods revealed that the five dfr genes accounted for 75–86% of trimethoprim resistance. The standard mPCR is useful and accessible for most laboratories, while the real-time mPCR requires additional equipment and expensive reagents, but is very convenient for high-throughput screening of large collections of bacterial isolates.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Clinical Microbiology and Infection - Volume 13, Issue 11, November 2007, Pages 1112–1118
نویسندگان
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