Development and application of a multiplex PCR assay for detection of the Clostridium perfringens enterotoxin-encoding genes cpe and becAB.
Clostridium perfringens causes food-borne gastroenteritis following the consumption of contaminated food by producing C. perfringens enterotoxin (CPE) in the intestines. Recently, we reported a novel enterotoxin, binary enterotoxin of C. perfringens (BEC) in C. perfringens isolates, which caused two disease outbreaks in Japan. Consequently, in the event of food poisoning outbreaks caused by C. perfringens, it is now necessary to screen for both the cpe and becAB genes by diagnostic PCR. Here, we present a simple multiplex PCR method for simultaneous detection of cpe, becAB and a C. perfringens control locus, phospholipase C (plc). Applying this method, we investigated the prevalence of cpe- or becAB-carrying C. perfringens strains in human stool and bovine rectum swab samples. Using a total of 169 isolates, we found that the percentage of becAB-carrying strains was very small (0.59%), one-tenth that of cpe-carrying strains. The simple method presented in this study with high specificity and sensitivity to C. perfringens will be a useful tool to survey the global prevalence of becAB-carrying C. perfringens strains.
Division of Bacteriology, Department of Infectious Disease, Osaka Prefectural Institute of Public Health, Osaka, Osaka, Japan; Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan. Electronic address: yonogi@iph.pref.osaka.jp.
Shinya
S
Yonogi
Division of Bacteriology, Department of Infectious Disease, Osaka Prefectural Institute of Public Health, Osaka, Osaka, Japan.
Masashi
M
Kanki
National Institute of Health Sciences, Tokyo, Tokyo, Japan.
Takahiro
T
Ohnishi
Matsubara Meat Hygiene Inspection Center, Matsubara, Osaka, Japan.
Masami
M
Shiono
Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
Tetsuya
T
Iida
Division of Bacteriology, Department of Infectious Disease, Osaka Prefectural Institute of Public Health, Osaka, Osaka, Japan.
Yuko
Y
Kumeda
D000818:Animals; D002417:Cattle; D003015:Clostridium Infections / Q000175:diagnosis / Q000382:microbiology; D003016:Clostridium perfringens / Q000235:genetics*; D004768:Enterotoxins / Q000235:genetics*; D005243:Feces / Q000382:microbiology*; D005759:Gastroenteritis / Q000175:diagnosis / Q000382:microbiology; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D007564:Japan; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004768:Enterotoxins; C035598:enterotoxin, Clostridium
BEC; CPE; Clostridium perfringens; Enterotoxin; Food-borne gastroenteritis; Multiplex PCR