猪鼻支原体套式PCR诊断方法的建立及应用

摘要

In order to develop a rapid detection mehthod for Mycoplamsa hyorhinis,two pairs of primerswere designed according to the P69 gene sequence of Mycoplamsa hyorhinis in the study,and the nested PCR method for Mycoplamsa hyorhinis detection was established.And then the specifity,sensitivity and clinical tests were carried out.The results showed that the sensitivity of the assay was reached to 1.4 ×10-5 ng/μL,and the detection method had no cross reaction with common bacterium and virus DNA from pig.In addition,twenty-three clinical specimens were detected by the nested PCR assay and the results demonstrated the positive rate was 73.9％.The nested PCR assay established in this study had the advantages of high specificity and high sensitivity,and could be used in clinical diagnosis of Mycoplamsa hyorhinis.%为建立猪鼻支原体快速诊断方法,根据猪鼻支原体P69基因序列设计2对引物,建立了猪鼻支原体套式PCR诊断方法,对建立的方法进行特异性、敏感性检测,同时进行临床检测.结果表明,应用建立的套式PCR方法对猪鼻支原体DNA的检出限为1.4×10-5 ng/μL,与常见感染猪的细菌、病毒均无交叉反应.利用建立的套式PCR方法对23份临床样品进行检测,阳性检出率为73.9％.本研究建立的猪鼻支原体套式PCR具有特异性强、敏感性高等优点,可用于猪鼻支原体的临床诊断.