کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2787654 1154336 2011 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Detection and identification of Vibrio parahaemolyticus by multiplex PCR and DNA–DNA hybridization on a microarray
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شناسی تکاملی
پیش نمایش صفحه اول مقاله
Detection and identification of Vibrio parahaemolyticus by multiplex PCR and DNA–DNA hybridization on a microarray
چکیده انگلیسی

In this paper, we developed a rapid and accurate method for the detection of Vibrio parahaemolyticus strains, using multiplex PCR and DNA–DNA hybridization. Multiplex PCR was used to simultaneously amplify three diagnostic genes (tlh, tdh and fla) that serve as molecular markers of V. parahaemolyticus. Biotinylated PCR products were hybridized to primers immobilized on a microarray, and detected by chemiluminesce with avidin–conjugated alkaline phosphatase. With this method, forty-five samples were tested. Eight known virulent strains (tlh+/tdh+/fla+) and four known avirulent strains (tlh+/tdh−/fla+) of the V. parahaemolyticus were successfully detected, and no non-specific hybridization and cross-hybridization reaction were found from fifteen closely-related strains (tlh−/tdh−/fla+) of the Vibrio spp. In addition, all the other eighteen strains of non-Vibrio bacteria (tlh−/tdh−/fla−) gave negative results. The DNA microarray successfully distinguished V. parahaemolyticus from other Vibrio spp. The results demonstrated that this was an efficient and robust method for identifying virulent strains of V. parahaemolyticus.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Genetics and Genomics - Volume 38, Issue 3, 20 March 2011, Pages 129–135
نویسندگان
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