title: Typing of Clostridium perfringens by in vitro amplification of toxin genes.
abstract: 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 alpha-toxin, beta-toxin, epsilon-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 alpha-toxin gene was obtained from direct enrichments of animal intestinal contents.
authors: Faculty of Veterinary Medicine, University of Liège, Sart Tilman, Belgium.
authors: G
authors: G
authors: Daube
authors: B
authors: B
authors: China
authors: P
authors: P
authors: Simon
authors: K
authors: K
authors: Hvala
authors: J
authors: J
authors: Mainil
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D003016:Clostridium perfringens / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D004269:DNA, Bacterial / Q000378:metabolism; D003670:Deer; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004767:Enterotoxemia / Q000382:microbiology; D005798:Genes, Bacterial / Q000235:genetics*; D006041:Goats; D006801:Humans; D007422:Intestines / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012756:Sheep; D013552:Swine
chemical_list: D001427:Bacterial Toxins; D017931:DNA Primers; D004269:DNA, Bacterial; D015252:Deoxyribonucleases, Type II Site-Specific