牛卵形巴贝斯虫不同靶基因PCR检测方法的比较  

Comparison of different PCR methods for the detection of Babesia ovata target gene

作　　者：田万年 李荣权 薛书江[2] 贾立军[2] 张守发[2] TIAN Wan-nian;LI Rong-quan;XUE Shu-Jiang;JIA Li-jun;ZHANG Shou-fa(College of Animal Science, Jilin Agricultural Seience and Technology College, Jilin 132101, China;College of Agriculture, Yanbian University, Yanji 133002 , China)

机构地区：[1]吉林农业科技学院动物科技学院,吉林吉林132101 [2]延边大学农学院,吉林延吉133002

出　　处：《中国兽医杂志》2018年第2期30-32,共3页Chinese Journal of Veterinary Medicine

基　　金：吉林省重点学科培育项目[吉农院合字(2015)第X028号]

摘　　要：旨在筛选出检测牛卵形巴贝斯虫特异、敏感的PCR方法。本试验以牛卵形巴贝斯虫18S rRNA、AMA-1和CCTη基因为靶基因进行PCR检测,从敏感性、特异性和临床检出率方面进行比较。结果显示,以18S rRNA基因的PCR方法敏感性最高,最小检出率为16 fg/μL;以CCTη为靶基因的PCR方法敏感性最低,检测量为1.6 pg/μL;而以顶膜抗原(AMA-1)为靶基因的PCR方法的最低检测量为160 fg/μL。三种靶基因均扩增不出牛瑟氏泰勒虫、牛巴贝斯虫和双芽巴贝斯虫基因片段。通过60份临床血液样本的检测结果表明,以18S rRNA基因设计引物的检出率最高,为30%(18/60),明显高于以AMA-1基因25%(15/60)和CCTη基因21.67%(13/60)。本试验为卵形巴贝斯虫病的诊断提供了更为敏感、特异的检测技术。To find a method which is more specific and sensitive to Babesia ovata, 18S rRNA, AMA - land CCTη gene of Babesia ovata were used as target genes and the sensitivity, specificity and detection rate of PCR of clinical samples were compared. The results showed that PCR method based on the 18S rRNA had the highest senssitivity,with minimum detectable amount of 16 fg/μL. While the CCTη gene had the lowest senssitivity ,with the minimum detectable amount of 1.6 pg/μL. The method showed no cross reaction with Theileria sergenti, Babesia bovis, and Babesia bigemina. The detection results for 60 clinical samples showed that the PCR method based on 18S rRNA gene 30% （18/60） had significantly higher detection rate than that based on AMA - 1 gene 25% （15/60） and CCTη gene 21.67% （13/60）. The data showed that the PCR method was specific and sensitive. This study provides a more sensitive and specific detection approach for Babesia ovata.

关 键 词：卵形巴贝斯虫 18S RRNA基因 AMA-1基因 CCTη基因 PCR方法 

分 类 号：S855.9[农业科学—临床兽医学]