关键词: 泰泽病原体, 兔, 实时荧光 PCR

Abstract: Objective To establish a real-time PCR method for diagnosis of Tyzzer’s organism in rabbits. Method One sets of primers and probes were designed according to the 16S rRNA sequence of Tyzzer’ s organism. The concentration of primers and probes was optimized, and the sensitivity, specificity and repeatability of this real-time PCR method were evaluated. At the same time, clinical samples were tested using this method and compared with nested PCR method reported before. Result A real-time PCR was developed to specifically detect Tyzzer ’ s Organism, detection limit of the positive plasmids were 8 copies/ μL. This method did not cross-react with various pathogens that should not be detected in laboratory animals. The coefficient of variation of intra- and inter-batch repetition was less than 3%, indicating that the repeatability of this method was good. According to the testing result of 70 clinical samples, it was found that the positive sample detection rate of this method was higher than that of the nested PCR method. Conclusion The real-time PCR method established in this study has good specificity, high sensitivity and good reproducibility, and was suitable for the detection of Tyzzer ’ s organism in rabbit fecal samples.

Key words: Tyzzer’ s organism, rabbit, real-time PCR