Kadra B, Guillou J P, Popoff M, Bourlioux P
Phylaxia Sanofi Co. Ltd, Budapest, Hungary.
FEMS Immunol Med Microbiol. 1999 Jul;24(3):259-66. doi: 10.1111/j.1574-695X.1999.tb01292.x.
A simple procedure was developed to identify toxitypes of Clostridium perfringens of different origins. Ninety strains of C. perfringens were identified by classical bacteriological methods, typing of the strains was done by a seroneutralisation test on mice. Production of enterotoxin was tested and all strains were analysed by PCR using gene of toxin alpha, gene of toxin E, gene of toxin beta and gene of enterotoxin. Simple amplification (amplifying one gene), and duplex and triplex amplification (amplifying two and three genes simultaneously) were performed. In the conditions of the experiment, the PCR method has proved efficacious. The specificity and sensitivity are excellent and superior to those of the classical methods. The prophylaxis of enterotoxaemia in animals is achieved by vaccination, the PCR technique can thus become a first-choice tool for the identification and typing of the C. perfringens strains which initiate these diseases. In turn, this would simplify the development of vaccines adapted to the epidemiological situation.
已开发出一种简单程序来鉴定不同来源的产气荚膜梭菌的毒素型别。采用经典细菌学方法鉴定了90株产气荚膜梭菌,通过对小鼠进行血清中和试验对菌株进行分型。检测了肠毒素的产生情况,并使用毒素α基因、毒素E基因、毒素β基因和肠毒素基因通过聚合酶链反应(PCR)对所有菌株进行分析。进行了简单扩增(扩增一个基因)以及双重和三重扩增(同时扩增两个和三个基因)。在本实验条件下,PCR方法已证明有效。其特异性和敏感性极佳,优于经典方法。动物肠毒血症的预防通过接种疫苗来实现,因此PCR技术可成为鉴定引发这些疾病的产气荚膜梭菌菌株及其分型的首选工具。相应地,这将简化适应流行病学情况的疫苗的研发。