Comparing the performance of conventional PCR, RTQ-PCR, and droplet digital PCR assays in detection of Shigella

Publication date: Available online 13 February 2020Source: Molecular and Cellular ProbesAuthor(s): Jin Yang, Nana Zhang, Jun Lv, Ping Zhu, Xing Pan, Jiaqingzi Hu, Wenfeng Wu, Shan Li, Hongtao LiAbstractThe 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 potentially reliable method for rapid and effective detection of Shigella spp.
Source: Molecular and Cellular Probes - Category: Molecular Biology Source Type: research