应用多重套式PCR检测鸡毒支原体和鸡滑液囊支原体

Detection of Mycoplasma synoviae and Mycoplasma gallisepticum using duplex PCR method

根据已发表的鸡毒和鸡滑液支原体血凝素基因序列pMGA和vlhA各设计两对引物,建立鉴别诊断两种支原体的多重套式PCR方法,对其进行温度条件、Ⅱ步模板浓度优化及特异性、敏感性实验。该方法在两步PCR后能特异性地扩增出MG(408 bp)和MS(688 bp)两个目的片段。应用于临床样品检测,与支原体分离、SPA检测比较结果PCR灵敏度高于病原分离。

On the basis of the hemagglutinin genes of pMGA fi＇om Mycoplasma gallisepticum （MG） and v1hA9 from Mycoplasma synoviae （MS）, a duplex PCR assay was developed and the reaction conditions including the reaction temperature, template concentration, specificity and sensitivity were optimized. The size of the PCR products was 408bp for MG and 688bp for MS. Three methods including duplex PCR, mycoplasma isolation and SPA were used to detect the samples from field. The results showed that the duplex PCR method was more sensitive than the methods of Myeoplasma isolation and SPA.