Sasaki Y, Yamamoto K, Kojima A, Tetsuka Y, Norimatsu M, Tamura Y
National Veterinary Assay Laboratory, Kokubunji, Tokyo, Japan.
J Vet Med Sci. 2000 Dec;62(12):1275-81. doi: 10.1292/jvms.62.1275.
Clostridium chauvoei causes blackleg, which is difficult to distinguish from the causative clostridia of malignant edema. Therefore, a single-step PCR system was developed for specific detection of C. chauvoei DNA using primers derived from the 16S-23S rDNA spacer region and partial 23S rDNA sequences. The specificity of the single-step PCR system was demonstrated by testing 37 strains of clostridia and 3 strains of other genera. A 509 bp PCR product, which is a C. choauvoei-specific PCR product, could be amplified from all of the C. chauvoei strains tested, but not from the other strains. Moreover, this single-step PCR system specifically detected C. chauvoei DNA in samples of muscle from mice 24 hr after inoculation with 100 spores of C. chauvoei, and in clinical materials from a cow affected with blackleg. These results suggest that our single-step PCR system may be useful for direct detection of C. chauvoei in culture and in clinical materials from animals affected with blackleg.
产气荚膜梭菌可引起黑腿病,这种疾病很难与恶性水肿的致病性梭菌区分开来。因此,利用源自16S-23S rDNA间隔区和部分23S rDNA序列的引物,开发了一种用于特异性检测产气荚膜梭菌DNA的一步法PCR系统。通过检测37株梭菌和3株其他属的菌株,证明了该一步法PCR系统的特异性。一个509 bp的PCR产物,即产气荚膜梭菌特异性PCR产物,可从所有检测的产气荚膜梭菌菌株中扩增出来,但不能从其他菌株中扩增出来。此外,该一步法PCR系统在接种100个产气荚膜梭菌孢子24小时后的小鼠肌肉样本以及患有黑腿病的奶牛的临床材料中特异性地检测到了产气荚膜梭菌DNA。这些结果表明,我们的一步法PCR系统可能有助于直接检测培养物和患有黑腿病动物的临床材料中的产气荚膜梭菌。