Yamagishi T, Sugitani K, Tanishima K, Nakamura S
Department of Laboratory Sciences, School of Health Sciences, Kanazawa University, Kodatsuno, Ishikawa, Japan.
Microbiol Immunol. 1997;41(4):295-9. doi: 10.1111/j.1348-0421.1997.tb01204.x.
In order to avoid the use of experimental animals, the polymerase chain reaction (PCR) method was applied to differentiate Clostridium perfringens into five toxin types. Twenty-two out of 23 strains tested produced the toxin(s) corresponding to the toxin gene(s) identified by PCR, and vice versa. Consequently, the gene typing was consistent with conventional typing by animal tests. Twenty-five strains were identified as types different from original ones by the PCR method as well as a toxin neutralization test. These findings suggest that the PCR method, which is easy and timesaving, is applicable to identify the toxin types of C. perfringens as an alternative to animal tests, and that beta-, epsilon- and iota-toxin genes might be lost by long-term preservation. The reasons why the strains lost the genes are discussed.
为避免使用实验动物，采用聚合酶链反应（PCR）方法将产气荚膜梭菌分为五种毒素类型。在检测的23株菌株中，有22株产生了与PCR鉴定的毒素基因相对应的毒素，反之亦然。因此，基因分型与传统的动物试验分型一致。通过PCR方法以及毒素中和试验，鉴定出25株菌株的类型与原始类型不同。这些结果表明，简单且省时的PCR方法可作为动物试验的替代方法用于鉴定产气荚膜梭菌的毒素类型，并且β-、ε-和ι-毒素基因可能会因长期保存而丢失。文中讨论了菌株基因丢失的原因。
