应用TaqMan荧光定量PCR快速检测鹿血中副结核分枝杆菌

摘要

为建立快速检测鹿血中副结核分枝杆菌(Mycobacterium avium subsp.pamtuberculosis,MAP)的荧光定量PCR方法,本研究根据GenBank中登录的MAP f57基因序列设计并合成引物及探针,并检测该方法的特异性和敏感性.试验结果显示,该方法具有良好的特异性,对MAP的检测灵敏度可以达到单个菌细胞.对长春地区采集的549份血清样品进行检测,结果显示阳性血清101份,阳性率达到18.4%.本研究结果表明,荧光定量PCR用于动物性产品MAP的检测具有快速、准确的特点.%For detection Mycobacterium avium subsp. paratuberculosis (MAP) in deer blood, a TaqMan PCR assay was developed. It targets and amplifies sequences from the f57 insertion element which is specific for this bacterium, the assay was tested against MAP and other mycobacterium strains. It is capable of detecting single cell. A total number of 549 serum samples collected from Changchun were tested by TaqMan PCR, the positive detection rates of these methods were 18.4％. The results showed that TaqMan PCR assay was rapid and accuracy for detection of MAP from infected deer products.