Improved multiplex real-time SYBR Green PCR assay for analysis of 24 target genes from 16 bacterial species in fecal DNA samples from patients with foodborne illnesses.

Improved multiplex real-time SYBR Green PCR assay for analysis of 24 target genes from 16 bacterial species in fecal DNA samples from patients with foodborne illnesses. Jpn J Infect Dis. 2015 Jul 10; Authors: Kawase J, Etoh Y, Ikeda T, Yamaguchi K, Watahiki M, Shima T, Kameyama M, Horikawa K, Fukushima H, Goto R, Shirabe K Abstract Here, we developed a new version of our screening system, Rapid Foodborne Bacterial Screening 24 (RFBS24), which can simultaneously detect 24 target genes of foodborne pathogens in fecal DNA samples. This new version (RFBS24 ver.5) detected all known stx2 subtypes, ETEC (STh), and Vibrio parahaemolyticus (trh2), which were not detected with the original RFBS24 system. The detection limits of the RFBS24 ver.5 system were approximately 5.6 × 10(-2) to 5.6 × 10(-5) ng DNA/reaction, significantly lower (10-100 times) than those of the RFBS24 system for the 22 target genes studied here. We also evaluated the new system using fecal DNA samples from patients infected with EHEC, Salmonella, and Campylobacter. The number of bacterial target genes detected using RFBS24 ver.5 was higher than that detected using RFBS24, and RFBS24 ver.5 combined with an Ultra Clean Fecal DNA Isolation Kit exhibited adequate performance (sensitivity and specificity: 89% and 100%, respectively, for Salmonella spp. and 100% and 83%, respectively, for Campylobacter jejuni) for rapid detection of the causative pathogen during foodborne o...
Source: Japanese Journal of Infectious Diseases - Category: Infectious Diseases Authors: Tags: Jpn J Infect Dis Source Type: research