利用聚合酶链反应技术检测牛结核杆菌病的研究

Detection of M bovis by Polymerase Chain Reaction

应用聚合酶链反应（ Ｐ Ｃ Ｒ） 技术检测牛结核分枝杆菌纯化 Ｄ Ｎ Ａ， 其敏感性为２５０ｆｇ 。所用引物序列对９种抗酸分枝杆菌 Ｄ Ｎ Ａ 进行扩增， 经琼脂糖凝胶电泳证实， 只有人型结核分枝杆菌、牛型结核分枝杆菌产生了３１７ｂｐ 的特异性扩增带。将 Ｐ Ｃ Ｒ 法与皮内变态反应试验（ Ｐ Ｐ Ｄ） 检测方法比较； ５４ 份血样标本中 Ｐ Ｃ Ｒ 的阳性率为１ ％ ， Ｐ Ｐ Ｄ 试验的阳性率为０ 。同时对奶样标本的检测与血样结果一致。结果表明， Ｐ Ｃ Ｒ 在直接检测患牛血样、奶样标本中显示出快速、敏感、特异的优点。为今后牛结核病的检测工作提供了一条新的途径。

The puried DNA from M bovis was detected by the polymerase chain reaction(PCR) The detection limit of the PCR assay for cultured mycobacteria was 250fg 9 species of mycobacter ia DNA were amplified by using The oligonucleotide primers,As expected,only M TB,M bovis showed the specific 317bp fragment by electrophoresed in 2 0%agarose gels PCR results were compared with PPD for the detection of M bovis in 54 blood specimens There was 1(1%)specimen that was positive for M bovis by PCR and 0(0%) that positive by PPD Meanwhile,The same result was obtained by using the specimen of blood as milk in one cow when pck amplified The result indicated that direct detection of M bovis in blood and milk specimen by PCR was rapid,sensitive,specific and provided a new approach for the diagnosis of boine tuberculosis in future