多房棘球绦虫粪抗原双抗体夹心ELISA检测方法的建立  

Development of a Double Antibody Sandwich ELISA Method for Detecting Fecal Antigen of Echinococcus multilocularis

作　　者：吴小霞 赵莉 王冰洁 班万里 穆尼拉·特列吾汗 马长丽 乌云花 布于其其格 陈云英 段兰利 岳城[1] 徐晶 艾沙江 张壮志 WU Xiao-xia;ZHAO Li;WANG Bing-jie;BAN Wan-li;Munila·Teliewuhan;MA Chang-li;Wu yunhua;Bu yuqiqige;CHEN Yun-ying;DUAN Lan-li;YUE Cheng;XU Jing;Aishajiang;ZHANG Zhuang-zhi(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi,Xinjiang,830000,China;Institute of Veterinary Medicine,Xinjiang Academy of Animal Sciences/Animal Clinical Medical Research Center of Xinjiang Academy of Animal Science,Urumqi,Xinjiang,830011,China;Tacheng District Animal Disease Control Center,Tacheng,Xinjiang,833400,China;Animal Husbandry and Veterinary Station,Bole City Bortala Mongolian Autonomous Prefecture,Bole,Xinjiang,833400,China;Animal Husbandry and Veterinary Station,Wenquan County,Boltala Mongolian Autonomous Prefecture,Boltala,Xinjiang,833000,China)

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830000 [2]新疆畜牧科学院兽医研究所/新疆畜牧科学院动物临床医学研究中心,新疆乌鲁木齐830011 [3]新疆塔城地区动物疫病预防控制中心,新疆塔城834300 [4]新疆博尔塔拉蒙古自治州博乐市畜牧兽医站,新疆博乐833400 [5]新疆博尔塔拉蒙古自治州温泉县畜牧兽医站,新疆博州833000

出　　处：《动物医学进展》2021年第8期1-6,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31560692)。

摘　　要：建立多房棘球绦虫粪抗原双抗体夹心ELISA检测方法,为犬感染多房棘球绦虫的早期诊断提供技术支撑。以5E10H5杂交瘤细胞株腹腔接种Balb/c鼠制备的腹水作为包被抗体,多房棘球绦虫成虫可溶性抗原免疫新西兰大白兔制备的多克隆抗体血清作为检测抗体,HRP标记的驴抗兔IgG作为二抗,建立双抗体夹心ELISA方法,检测试验犬(感染多房棘球绦虫)和阴性对照犬犬粪。结果显示,多房棘球绦虫成虫可溶性抗原具有良好的抗原性并能产生高效价抗体;对该方法的灵敏度进行检测,阳性粪样稀释至1:10000时仍显示为阳性;在感染动态分析中,该方法最早可在犬感染72 h后,最迟6 d后检测到多房棘球绦虫粪抗原。表明建立的方法可用于诊断犬多房棘球绦虫感染,为后期研制犬多房棘球绦虫粪抗原双抗体夹心ELISA检测试剂盒奠定了基础。The aim of the study was to develop a double antibody sandwich ELISA method for detection of Echinococcus multilocularis fecal antigen, and to provide technical support for the early diagnosis of Echinococcus multilocularis infection in dogs.The method was established using the ascites prepared by intraperitoneal inoculation of the 5 E10 H5 hybridoma cell line into Balb/c mice as coating antibody, the polyclonal antibody serum prepared by immunizing New Zealand white rabbits with soluble antigens of Echinococcus multilocularis adults as the detection antibody, and the HRP-labeled donkey anti-rabbit IgG as the secondary antibody, to detect test dogs(infected with Echinococcus multilocularis) and negative control dog feces.The results showed that: the soluble antigen of Echinococcus multilocularis adults has good antigenicity and can produce high-efficiency antibodies;The sensitivity of the method is detected, and the positive feces diluted to 1∶10 000 still show positive;In the dynamic analysis of infection, this method can detect Echinococcus multilocularis fecal antigen as early as 72 hours after the dog is infected, and 6 days later at the latest.It showed that the method established in this study can be used to diagnose Echinococcus multilocularis infection in dogs, laying a foundation for the later development of a double antibody sandwich ELISA test kit for the canine Echinococcus multilocularis fecal antigen.

关 键 词：多房棘球绦虫 双抗体夹心ELISA 粪抗原 

分 类 号：S852.734[农业科学—基础兽医学]