Enterotoxin gene detection by polymerase chain reaction and genotyping of Clostridium perfringens by pulsed-field gel electrophoresis

Abstract

Enterotoxin gene detection in 106 Clostridium perfringens isolates including 80 isolates from food and water, 20 isolates from stool of non-admitted patients with diarrhea, one isolate from stool of non-admitted patient with diarrhea and 5 isolates from food poisoning outbreaks in Japan was performed using polymerase chain reaction (PCR) technique. None of the isolates from food and water and from food and water and from stool of the admitted patients had enterotoxin gene while the isolate from stool of one non-admitted patient as well as all 5 food poisoning isolates from Japan had enterotoxin gene. Reverse passive latex agglutination (RPLA) was also used to detect CPE production in vitro. It was indicated that cpe gene detection by PCR and RPLA did not provide correlated results in the detection of enterotoxigenic isolates. Thus, RPLA required in vitro sporulation and not all isolates could sporulate in the laboratory conditions used in this study. All C.perfringens isolates were discriminated into 88 different pulsotypes using pulsed-field gel electrophoresis (PFGE) of the Sma I restricted chromosomal fragments. One isolate from non-admitted patient with diarrhea was in the same pulsotype as the two food poisoning isolates from Japan which was Pulsotype 85