Comparing the performance of conventional PCR,RTQ-PCR,and droplet digital PCR assays in detection of Shigella |
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| Institution: | 1. Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran;2. Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran;1. Department of Clinical Laboratory, The Second People''s Hospital of Zhuhai, Zhuhai 519020, Guangdong, PR China;2. State Key Laboratory of Diarrhea Disease Detection, Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai 519020, Guangdong, PR China |
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| Abstract: | The incidence of foodborne infections caused by Shigella spp. is still very high in every year, which poses a great potential threat to public health. Conventional quantification methods based on culture techniques, biochemical, and serological identification are time-consuming and labor-intensive. To develop a more rapid and efficient detection method of Shigella spp., we compared the sensitivity and specificity of three different polymerase chain reaction (PCR) methods, including conventional PCR, quantitative real-time PCR (RTQ-PCR), and droplet digital PCR (ddPCR). Our results indicated that ddPCR method exhibited higher sensitivity, and the limit of detection was 10?5 ng/μl for genomic DNA templates, 10?1 cfu/ml for Shigella bacteria culture. In addition, we found that ddPCR was a time-saving method, which required a shorter pre-culturing time. Collectively, ddPCR assay was a reliable method for rapid and effective detection of Shigella spp. |
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| Keywords: | Foodborne infections Droplet digital PCR Quantitative real-time PCR Conventional PCR |
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