[Study of real-time PCR assays for rapid detection of food-borne pathogens].
A duplex real-time SYBR Green LightCycler PCR (LC-PCR) assay with DNA extraction using QIAamp DNA Stool Minikit was evaluated for detection of 8 of 19 species of food-borne pathogens, including Plesiomonas shigelloides, Providencia alcalifaciens, in five stool specimens. The time frame was within 2h or less. The protocol used the same LC-PCR with 22 pairs of specific primers. The rapid amplification and reliable detection of specific genes were determined by this LC-PCR assay from 10 cases of food-borne outbreaks in Shimane Prefecture from 2002 to 2005. These cases included Campylobacter jejuni (4), Clostridium perfringens (2), enteropathogenic Escherichia coli and astA positive E. coli (1), and astA positive E. coli, enterohemorrhagic E. coli 026, and Bacillus cereus (1 each). Rapid amplification and reliable detection of specific genes of food-or water-borne pathogenic bacteria in fecal samples should facilitate the diagnosis and management of food-borne outbreaks.
Fukushima
Hiroshi
H
Shimane Prefectural Institute of Public Health and Environmental Science, 582 Nishihamasada, Matsue, Shimane 690-0122, Japan.
Tsunomori
Yoshie
Y
D001483:Base Sequence; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification; D003016:Clostridium perfringens / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics*; D003967:Diarrhea / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D005516:Food Microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011933:Reagent Kits, Diagnostic / Q000592:standards
D017931:DNA Primers; D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic