JAPANESE JOURNAL OF FOOD MICROBIOLOGY
Online ISSN : 1884-5754
Print ISSN : 0910-8637
ISSN-L : 0910-8637
Rapid Detection of Clostridium perfringens Type A Enterotoxin Gene by Using the Polymerase Chain Reaction
Katsuyuki ISHIMURATakayuki KAYASHIMAKazumasa KURATAFumiaki ITOHKiyoshi NAKANOTakeaki MATSUISHITakeo OGINO
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1993 Volume 10 Issue 1 Pages 35-41

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Abstract

Synthetic oligonucleotide primers were used in the polymerase chain reaction (PCR) technique to detect the gene encoding Clostridium perfringens type A enterotoxin (CPE). Sequences of the primers are 5'-ATCCTTGATTTAGCTGCTGC-3' and 5'-ACAAGAACATATTGTCCGGC-3', which are constructed to amplify a 302-base-pair (bp) region of the CPE gene. All CPE-producing C. perfringens strains tested, such as C. perfringens NCTC8239, gave an amplified DNA fragment, while non-CPE producing C. perfringens strains gave no amplification. The amplified DNA fragment was recognized at a molecular size of about 360-bp on polyacrylamide gel electrophoresis (PAGE) at room temperature, but migrated about 302-bp on PAGE at 53°C. Each cleavage pattern of the amplified DNA with four restriction endonucleases, Dde I, Hinf I, Mbo I, and Hind III, was consistent with the patterns expected from the published sequence of the CPE gene.
In outbreaks, the enterotoxigenicity of C. perfringens isolates from patients, food handlers, or incriminated foods could rapidly be judged by the PCR method, the results of which were consistent with those of the culture and the immunological method. Thus, these results show that the PCR method provides a simple and rapid tool for the detection of the potentially enterotoxigenic C. perfringens, and furthermore, suggest that CPE is not a common structural component of the C. perfringens spore coat.

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© Japanese Society Food Microbiology
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