驽巴贝斯虫PCR检测方法的建立及应用

根据驽巴贝斯虫BC-48基因序列,设计并合成1对特异性引物,建立了驽巴贝斯虫PCR检测方法。试验扩增出610 bp的基因片段,其序列与GenBank上驽巴贝斯虫BC-48基因同源性为96.7%。而对新孢子虫、弓形虫、马巴贝斯虫的基因组DNA没有扩增带出现。对驽巴贝斯虫基因组DNA的最小检测量为2.812 fg/μL。通过对35份临床样品的检测,阳性率为20%。同时与血液涂片染色镜检进行了比较,结果表明,PCR检测方法准确、敏感、特异。

Establishment of A PCR Assay Detecting Babesia caballi

One pair of specific primers was designed based on the BC-48 gene sequence of Babesia caballi, and then a PCR assay was established for detecting Babesia caballi. A 610 bp sequence was amplified by PCR, and the homology with the sequence from GenBank was 96.7%. No PCR products were amplified from purified DNA of Sporozoan tachyzoites, Toxoplasma tachyzoites and Theileria sergenti. This method can detect less than 2. 812fg/t,L of the genome DNA of Babesia caballi. Using this method, the 35 blood samples were successfully detected and the posi tive rate was 20 %. The results compared with staining method showed that the PCR method was precise, sensitive and specific.