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Typing of Clostridium perfringens by in vitro amplification of toxin genes
Author(s) -
Daube G.,
China B.,
Simon P.,
Hvala K.,
Mainil J.
Publication year - 1994
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1994.tb02815.x
Subject(s) - clostridium perfringens , toxin , enterotoxin , typing , polymerase chain reaction , agarose gel electrophoresis , biology , microbiology and biotechnology , gene , antiserum , clostridium , agarose , genetics , escherichia coli , bacteria , antibody
G. DAUBE, B. CHINA, P. SIMON, K. HVALA AND J. MAINIL. 1994. The strains of Clostridium perfringens are classified according to major toxins produced. Classically, this determination involves the seroneutralization of their lethal effect in mice. However, this method requires specific antisera and a large number of mice. In this work, a new typing method was developed based on the amplification of toxin genes by polymerase chain reaction (PCR). By combination of several pairs of primers, the toxinotype of a Cl. perfringens strain was determined by looking at the pattern of bands on an agarose gel electrophoresis. This mixture contained primers amplifying simultaneously a part of α‐toxin, α‐toxin, β‐toxin and enterotoxin genes. In order to distinguish between toxinotype A and E, the *** l ‐toxin gene fragment must be amplified in a separate PCR reaction. Moreover, with the primers combination, in most cases, a PCR product corresponding to the α‐toxin gene was obtained from direct enrichments of animal intestinal contents.