羊泰勒虫不同PCR检测方法的比较

Comparison of different PCR methods for the detection of Theileria

为寻求快速、有效检测羊泰勒虫的PCR方法,本试验建立了检测羊泰勒虫18SrRNA基因和表面蛋白基因的两种常规PCR方法和一种检测18SrRNA基因的半套式PCR方法,并从其敏感性和临床样本检出率等方面进行了比较。结果显示:上述方法检测羊泰勒虫基因组DNA的最小检测量分别为1.6fg/μL、16fg/μL和0.016fg/μL;检测临床样本阳性检出率分别为31.37%(16/51)、17.64%(9/51)和45.10%(23/51)。3种方法中,检测18SrRNA基因的半套式PCR方法敏感性和临床样本检出率最高,其次为检测18SrRNA基因的常规PCR方法,最后为检测表面蛋白基因的常规PCR方法。结果说明,所建立的半套式PCR方法是一种较好的羊泰勒虫检测方法。

To seek a rapid and valid PCR method for the detection of Theileria, we established two kinds of routine PCR for 18S rRNA gene and surface protein gene, respectively, and a semi-nested PCR for 18S rRNA gene. Their sensitivities and positive rates of the samples were detected and compared. The results showed that using the semi-nested PCR and the routine PCR to detect 18S rRNA and the routine PCR to detect surface protein gene, the minimum detection amounts were estimated 0. 016 fg/μL, 1.6 fg/μL and 16 fg/μL, respectively; the positive rates were 45. 10% （23/51）, 31.37% （16/51） and 17.64% （9/51）, respectively. It suggested that the semi-nested PCR had a higher sensitivity for Theilerla than the other two routine PCR, and could be used to detect Theileria in clinical samples.