羊泰勒虫PCR检测方法的建立和初步应用

利用羊泰勒虫18SrRNA基因的序列特点,设计合成种特异性引物,建立羊泰勒虫PCR检测方法,该方法能特异性扩增398bp的羊泰勒虫18SrRNA基因片段,而对羊巴贝斯虫、羊无浆体、牛环形泰勒虫和牛伊氏锥虫的基因组DNA没有扩增带出现。对羊泰勒虫基因组DNA的最小检测量为0.12fgDNA。通过检测124份临床样品,24份为羊泰勒虫感染阳性,其余为阴性。结果表明,建立的PCR检测方法具有极高的敏感性和特异性,可用于羊泰勒虫病和临床健康带虫羊的诊断。

Development and Preliminary Application of PCR Assay Based on 18S Rrna Gene for Detection of Theileria Sp. Infection

Based on the sequence data of 18S rRNA gene of Theileria sp,a pair of T.sp-specific primers were designed.These primers selectively amplified a 398bpDNA fragment of the 18S rRNA gene of Theileria sp.No PCR products were amplified from purified DNA of Babesia sp,Anaplasma ovis,Theileria annulata and Trypanosoma.It was estimated that as few as 0.12fg Theileria SP genomic DNA was detectable by PCR.Using this method ,124 blood samples from different area were successfully detected and 24 samples were positive with Theileria sp infection,the others were negative.The results showed that the PCR method developed was an effective tool for diagnosing Theileria SP infection in sheep and goats with high sensitivity and specificity.It will be useful for detecting the acute Theileria SP and latent infected carrier animals.