摘要
为特异性检测羊感染仰口线虫的情况,基于仰口线虫ITS基因序列设计特异性引物,进行退火温度及反应条件的优化,建立其属特异性PCR检测方法。应用该方法对仰口线虫、食道口线虫、毛尾线虫、捻转血矛线虫、细颈线虫、奥斯特线虫、夏伯特线虫、筒线虫、毛圆线虫的DNA样本进行扩增,仅能特异性扩增出仰口线虫的目的条带;该方法成功从仰口线虫中扩增出约500 bp的特异性片段,最低检测浓度为3.6×10^(3)copies/μL;该PCR检测方法对临床奶山羊粪便样品的检测结果显示,羊粪便样品中仰口线虫卵的阳性率为10%,且与显微镜镜检所得的结果一致。结果表明,建立的仰口线虫PCR检测方法敏感性较高且特异性良好,可以应用于临床羊粪便样品中仰口线虫的虫卵检测,为羊仰口线虫病的诊断与防治提供了一种技术支持。
In order to specifically detect the infection of Bunostomum,a genus-specific PCR detection method was established by designing specific primers based on the ITS gene sequence of Bunostomum and optimizing annealing temperature and reaction conditions,which was used to amplify the DNA samples of Bunostomum,Oesophagostoma,Trichuris,Haemonchus contortus,Nematodirus,Ostertagia,Chabertia,Gongylonema and Trichostrongylus.The result showed only the target DNA band of Bunostomum could be amplified.This method successfully amplified a specific fragment of about 500 bp from Bunostomum,with the minimum detection concentration of 3.6×10^(3)copies/μL.The detection of fresh dairy goat feces using optimized PCR method showed that the positive rate of Bunostomum in goat feces samples was 10%,and the positive rate was consistent with that of microscopy.The above-mentioned results indicated that the PCR method established in this study exhibits a high sensitivity and good specificity,which could be applied to the clinical detection of Bunostomum eggs in goat feces,providing technical support for the diagnosis and control of Bunostomum disease in goats.
作者
周璐露
要慧中
刘天缘
张佳琳
马丽
任洪英
杨钰莹
林青
ZHOU Lu-lu;YAO Hui-zhong;LIU Tian-yuan;ZHANG Jia-lin;MA Li;REN Hong-ying;YANG Yu-ying;LIN Qing(College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi,712100,China;Agriculture,Animal Husbandry,Rural Areas and Science and Technology Bureau of Luhuo County,Luhuo,Sichuan,730046,China)
出处
《动物医学进展》
北大核心
2024年第6期15-19,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(畜牧专项资金-XN14
畜牧防疫专项资金-XNDY2205)。
