牛瑟氏泰勒虫不同靶基因PCR检测方法的比较

Comparison of different PCR methods for the target genes of bovine Theileria sergenti

为筛选出检测牛瑟氏泰勒虫更为特异、敏感的PCR方法,本试验以牛瑟氏泰勒虫p23和p33基因、HSP70基因及18S rRNA基因为靶基因进行PCR检测,并从其敏感性、特异性和临床样本检出率等方面进行了比较。结果显示,4种靶基因引物对牛瑟氏泰勒虫的检测均有较高的特异性,当牛瑟氏泰勒虫基因组DNA浓度为127 ng/μL时,p23、p33、HSP70及18S rRNA4种基因的最小检测量分别为1×105、1×105、1×104和1×106copies/μL;检测临床样本阳性检出率分别为30.19%(16/53)、39.62%(21/53)、47.17%(25/53)和54.72%(29/53)。表明以18SrRNA基因为靶基因的PCR方法从敏感性和临床检出率上明显优于其他3种基因。

To develop a more specific and sensitive PCR method for bovine Theileria sergenti, p23, p33, HSP70 and 18S rRNA genes of T. sergenti were selected as the target genes, while the sensitivity and specificity of the methods and detection rates of clinical samples were compared. The results showed that four kinds of primers for the target genes of bovine T. sergenti had high specificity. When the concentration of T. sergenti genomic DNA was 127 ng/μL, the minimum detectable amount of p23, p33, HSP70 and 18S rRNA genes were 1 ×10^5, 1 × 10^5, 1× 10^4 and 1 × 10^6 copies/p.L, respectively. The positive rates of clinical samples were 30. 19% （16/53）, 39. 62% （21/53）, 47. 17% （25/53） and 54. 72% （29/53） , respectively. It suggested that the 18S rRNA gene as a target gene for PCR method was more specific and sensitive than the other three genes.