Establishmen of Real-time PCR for detecting Bibersteinia trehalosi for rpop gene

Abstract

Bibersteinia trehalosi is a zoonosis bacterial species, There are few reports of the disease in our country, and there is no fast, sensitive and efficient detection method have been developded. Therefore, the Quantitative Real-time PCR (q-PCR) for detecting Bibersteinia trehalosi has been established, and the specific primers desigened based on the conserved sequence of Bibersteinia trehalosi rpoβ gene, the method was constucted by optimizing reaction conditions and then subjected the sensitivity, specificity, reproducibility, and clinical samples test. The results indicated a good linear relationship when the concentration of standard substance ranged between 1.20×101~1.20×105 copies/μL. Inaddition, the minimum detectable concentration was 1.20×10−2 copies/μL. There are no cross reaction with 20 kinds of bacteria and viruses. All of the CV values of intra-groups and intra-groups were less than 3%.The results showed that the method has the advantages of high sensitivity, strong specificity, good stability, high accuracy and rapid detection. It can be used for early diagnosis of infections in Bibersteinia trehalosi, as well as rapid detection and quantitative analysis of samples.