摘要
为了能对羊源食道口线虫病进行特异性诊断,基于食道口线虫ITS基因序列设计特异性引物,建立食道口线虫的特异性PCR检测方法,对其特异性、敏感性以及在临床羊粪便样本中食道口线虫虫卵的检测效果进行验证。结果显示,在对食道口线虫、仰口线虫、毛尾线虫、捻转血矛线虫等9种线虫的阳性DNA模板进行扩增时,只有在食道口线虫DNA阳性模板中扩增出650 bp的目的条带,而在其他线虫的DNA模板中无条带出现;将食道口线虫原始质粒模板进行10倍梯度稀释后进行敏感性检测,该PCR最低检出浓度为90 copies/μL;利用所建立的PCR检测方法对临床200份羊粪便样本进行检测,结果检出食道口线虫的阳性率为32%,高于显微镜镜检的阳性检测结果(阳性率为31%),但二者无显著性差异。研究表明,该研究所建立的食道口线虫PCR检测方法能够特异性地检测食道口线虫及其虫卵的阳性DNA样本,且敏感性良好,该方法为食道口线虫的种类鉴别以及食道口线虫病的诊断提供了一种可供选择的技术。
In order to make specific diagnosis of oesophagostomiasis of goats,specific primers based on ITS gene sequence were designed to establish a specific PCR detection method for Oesophagostomum,and its specificity,sensitivity and detection effect of Oesophagostomum eggs in clinical goat feces samples were verified.The results showed that the 650 bp target band was amplified only in the positive DNA template of Oesophagostomum,in other nematodes including Bunostomum,Trichocephalus and Haemonchus contortus,no target band was observed.The original plasmid template of Oesophagostomum was diluted 10-fold and the sensitivity was detected,showing the minimum detectable concentration of PCR was 90 copies/μL.The positive rate of Oesophagostomum detected by PCR was 32%in the 200 feces samples collected clinically,higher than that by microscopy(31%),but there was no significant difference between the two methods.In conclusion,the PCR method established in this study can specifically detect positive DNA samples of Oesophagostomum and its eggs,and has good sensitivity.This method provides an alternative technical support for the species identification of Oesophagostomum and the diagnosis of oesophagostomiasis.
作者
刘天缘
要慧中
周璐露
康煜坤
林青
LIU Tian-yuan;YAO Hui-zhong;ZHOU Lu-lu;KANG Yu-kun;LIN Qing(College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi 712100,China;Lvliang Agricultural Comprehensive Administrative Law Enforcement Team,Lvliang,Shanxi 033000,China;Key Laboratory of Ruminant Disease Prevention and Control(West),Ministry of Agriculture and Rural Affairs,Yangling,Shaanxi 712100,China)
出处
《动物医学进展》
北大核心
2024年第11期52-56,共5页
Progress In Veterinary Medicine
基金
陕西省农业农村厅省级农业专项资金项目(畜牧专项XN14
畜牧防疫专项XNDY2205)。
关键词
食道口线虫
内转录间隔区
聚合酶链反应
羊
Oesophagostomum
internal transcriptional spacer
polymerase chain reaction
goat
