Uzal F A, Plumb J J, Blackall L L, Kelly W R
Department of Veterinary Pathology, University of Queensland, Brisbane, Australia.
Lett Appl Microbiol. 1997 Nov;25(5):339-44. doi: 10.1046/j.1472-765x.1997.00247.x.
A polymerase chain reaction (PCR) was used to identify the genes encoding the major toxins of Clostridium perfringens in faeces of goats. When pure cultures of Cl. perfringens types A, B, C, D and E were used as templates in the PCR, amplicons were observed on the agarose gel as bands at approximately the 247 (alpha primers), 1025 (beta primers), 403 (epsilon primers) and 298 (iota primers) bp level of the DNA marker. When used to identify different types of Cl. perfringens in samples artificially spiked with these micro-organisms, the PCR detected as few as 1-1.5 x 10(2) cfu g-1 of the five types of Cl. perfringens tested. The PCR technique allowed the identification and typing of Cl. perfringens strains in faeces of goats, without recourse to other techniques such as the mouse neutralization test.
采用聚合酶链反应(PCR)鉴定山羊粪便中产气荚膜梭菌主要毒素的编码基因。当产气荚膜梭菌A、B、C、D和E型的纯培养物用作PCR模板时,在琼脂糖凝胶上观察到扩增子,其条带位于DNA标记物约247bp(α引物)、1025bp(β引物)、403bp(ε引物)和298bp(ι引物)水平处。当用于鉴定人工接种这些微生物的样品中的不同类型产气荚膜梭菌时,PCR能检测到低至1 - 1.5×10² cfu g⁻¹的所测试的五种产气荚膜梭菌类型。PCR技术能够在不借助其他技术(如小鼠中和试验)的情况下,对山羊粪便中的产气荚膜梭菌菌株进行鉴定和分型。