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

Mol Cell Probes. 2020 Jun:51:101531. doi: 10.1016/j.mcp.2020.101531. Epub 2020 Feb 13.

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.

Keywords: Conventional PCR; Droplet digital PCR; Foodborne infections; Quantitative real-time PCR; Shigella spp. detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers
  • Dysentery, Bacillary / diagnosis*
  • Dysentery, Bacillary / microbiology
  • Feces / microbiology
  • Limit of Detection
  • Mice
  • RAW 264.7 Cells
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Shigella / genetics*
  • Shigella / isolation & purification*
  • Shigella / pathogenicity

Substances

  • DNA Primers