Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, State Key Laboratory Breeding Base of Eco-Environment and Bio-Resource, Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, School of Life Sciences, Southwest University, Beibei, 400715, Chongqing, China.
Institute of Infection and Immunity, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, China.
Mol Cell Probes. 2020 Jun;51:101531. doi: 10.1016/j.mcp.2020.101531. Epub 2020 Feb 13.
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 ng/μl for genomic DNA templates, 10 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.
志贺氏菌引起的食源性感染的发病率仍然很高，每年都在增加，这对公众健康构成了巨大的潜在威胁。基于培养技术、生化和血清学鉴定的传统定量方法既耗时又费力。为了开发一种更快速、高效的志贺氏菌检测方法，我们比较了三种不同聚合酶链反应（PCR）方法的灵敏度和特异性，包括常规 PCR、实时定量 PCR（RTQ-PCR）和数字液滴 PCR（ddPCR）。我们的结果表明，ddPCR 方法具有更高的灵敏度，基因组 DNA 模板的检测限为 10ng/μl，志贺氏菌培养物的检测限为 10cfu/ml。此外，我们发现 ddPCR 是一种节省时间的方法，它需要更短的预培养时间。总的来说，ddPCR 检测方法是一种快速、有效的检测志贺氏菌的可靠方法。
