RRC ID 48483
Author Kawase J, Etoh Y, Ikeda T, Yamaguchi K, Watahiki M, Shima T, Kameyama M, Horikawa K, Fukushima H, Goto R, Shirabe K.
Title An 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.
Journal Jpn. J. Infect. Dis.
Abstract Here, we developed a new version of our original screening system (Rapid Foodborne Bacterial Screening 24; RFBS24), which can simultaneously detect 24 genes of foodborne pathogens in fecal DNA samples. This new version (RFBS24 ver. 5) detected all known stx2 subtypes, enterotoxigenic Escherichia coli (STh genotype), and Vibrio parahaemolyticus (trh2), which were not detected by the original RFBS24 assay. The detection limits of RFBS24 ver. 5 were approximately 5.6 × 10(-2)-5.6 × 10(-5) (ng DNA)/reaction, significantly lower (10- to 100-fold) than those of the original RFBS24 for the 22 target genes analyzed here. We also tested the new assay on fecal DNA samples from patients infected with Salmonella, Campylobacter, or enterohemorrhagic E. coli. The number of bacterial target genes detected by RFBS24 ver. 5 was greater than that detected by RFBS24. RFBS24 ver. 5 combined with an Ultra Clean Fecal DNA Isolation Kit showed adequate performance (sensitivity and specificity 89% and 100%, respectively, for Salmonella spp. and 100% and 83%, respectively, for Campylobacter jejuni) in terms of rapid detection of a causative pathogen during foodborne-illness outbreaks. Thus, RFBS24 ver. 5 is more useful than the previous assay system for detection of foodborne pathogens and offers quick simultaneous analysis of many targets and thus facilitates rapid dissemination of information to public health officials.
Volume 69(3)
Pages 191-201
Published 2016-5-20
DOI 10.7883/yoken.JJID.2015.027
PMID 26166499
MeSH Bacterial Typing Techniques Campylobacter jejuni / genetics Campylobacter jejuni / isolation & purification DNA Primers / chemistry DNA, Bacterial / genetics* Escherichia coli O157 / genetics Escherichia coli O157 / isolation & purification Feces / microbiology Fluorescent Dyes Foodborne Diseases / diagnosis* Foodborne Diseases / microbiology High-Throughput Screening Assays Humans Limit of Detection Listeria monocytogenes / genetics Listeria monocytogenes / isolation & purification Multiplex Polymerase Chain Reaction / methods* Organic Chemicals Real-Time Polymerase Chain Reaction / methods* Salmonella / genetics Salmonella / isolation & purification Vibrio parahaemolyticus / genetics Vibrio parahaemolyticus / isolation & purification
IF 1.014
Times Cited 0
General Microbes JCM 7674 JCM 2529