马驽巴贝斯虫ddPCR 检测方法的建立

Development of ddPCR for Detection of Babesia caballi

马驽巴贝斯虫(Babesia caballi)病是马属动物的一种血液原虫病,被农业农村部列为二类动物疫病。本研究旨在建立一种针对马驽巴贝斯虫病的ddPCR检测技术。利用马驽巴贝斯虫特异性引物及探针对反应体系进行优化,测试了方法的特异性、稳定性及灵敏性,对30份可疑样品进行了初步检测。结果显示,该方法具有较好的稳定性,最低定量检测限可达3.3 copies/μL,对30份样品进行检测,马驽巴贝斯虫病阳性率为63.33%(19/30)。表明所建立的方法可成功用于马驽巴贝斯虫的检测。

Babesia caballi is one of the etiological agents of equine babesiosis,which was listed as the second-class epidemic disease by Ministry of Agriculture and Rural Affairs of China.In this study,a ddPCR method was developed for detection of B.caballi infection by using a pair of specific primers and a probe.This method was tested for optimal specificity,stability and sensitivity.The results showed that this method was significantly reproducible,and its detection limit was 3.3 copies/μL,In addition,30 suspicious blood samples were tested using this method and the positive rate was 63.33%(19/30)for B.caballi.