基于TashHN基因的牛环形泰勒虫病PCR诊断方法的建立  

Establishment of a PCR Diagnostic Method for Bovine Theileria annulata Based on TashHN Gene

作　　者：周子成 赵洪喜[1] Zhou Zicheng;Zhao Hongxi(School of Agriculture,Ningxia University,Yinchuan 750000,China)

机构地区：[1]宁夏大学农学院,宁夏银川750000

出　　处：《农业科学研究》2024年第1期74-78,共5页Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31760727);宁夏自然科学基金项目(2021AAC03011)。

摘　　要：根据Genbank已发表的环形泰勒虫TashHN基因序列设计合成引物,以此建立基于环形泰勒虫TashHN基因的PCR检测方法,对该方法的特异性、敏感性、稳定性进行验证,并用建立的方法对宁夏固原地区各县共计135份牛血样进行检测。结果表明:建立的基于TashHN的PCR检测方法与中华泰勒虫、东方泰勒虫、牛巴贝斯虫、双芽巴贝斯虫均无交叉反应;可检测的最低模板浓度为1.64×10^(2) copies/μL;环形泰勒虫的DNA在-20℃冰箱放置8个月后仍可检测出目的条带;该方法利用已有文献中的PCR方法进行一致性检测,从135份牛血样中检测出5份阳性,说明该方法准确可靠。因此,试验建立的基于环形泰勒虫TashHN基因的PCR检测方法具有特异性强、敏感性高、稳定性强的特点,适用于临床上环形泰勒虫早期感染检测以及流行病学的调查。The synthetic primers were designed and synthesized based on the TashHN gene sequence of Theileria annulata published in Genbank,and a PCR detection method based on the TashHN gene was established.The specificity,sensitivity,and stability of this method were tested,and the established method was used to detect 135 bovine blood samples from various counties in the Guyuan City of Ningxia.The results showed that the established PCR detection method based on the TashHN gene had no cross-reaction with T.sinensis,T.orientalis,Babesia bovis,and B.bigemina;the lowest detectable template concentration was 1.64×102 copies/μL;and the DNA of Theileria annulata could still be detected after being stored in a-20℃freezer for eight months.Five positive samples were detected out of 135 bovine blood samples from Guyuan using this method,which were consistent with those obtained using PCR methods described in existing literature,indicating that the method is accurate and reliable.The established PCR detection method based on the TashHN gene of T.annulata in this experiment has high specificity,sensitivity,and stability,and is suitable for early infection detection of T.annulata in clinical settings and for epidemiological surveys.

关 键 词：牛 环形泰勒虫 PCR 诊断方法 

分 类 号：S854.4[农业科学—临床兽医学]