禽呼肠孤病毒抗体检测胶体金试纸条的研制  

Preparation of Colloidal Gold Strips for Detection of Avian Reovirus Antibody

作　　者：杨冰怡 谢芝勋 黄娇玲 韦悠 谢丽基 谢志勤 罗思思 任红玉 YANG Bing-yi;XIE Zhi-xun;HUANG Jiao-ling;WEY You;XIE Li-ji;XIE Zhi-qin;LUO si-si;REN Hong-yu(College of Animal Science and Technology,Guangxi University,Nanning,Guangxi,530004,China;Guangxi Key Laboratory of Veterinary Biotechnology,Key Laboratory of China(Guangxi)-ASEAN Cross-border Animal Disease Prevention and Control,Ministry of Agriculture and Rural Affairs of China,Guangxi Veterinary Research Institute,Nanning,Guangxi,530004,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]广西壮族自治区兽医研究所,广西兽医生物技术重点实验室,农业农村部中国广西-东盟跨境动物疫病防控重点实验室,广西南宁530001

出　　处：《动物医学进展》2025年第3期33-38,共6页Progress In Veterinary Medicine

基　　金：广西科技重大专项(桂科AA23062050);广西科技基地和人才专项(AD17195083);广西八桂学者专项(2019A50)。

摘　　要：为建立禽呼肠孤病毒抗体的特异快速检测方法,用兔抗鸡IgG标记胶体金制作金标垫,将禽呼肠孤病毒(ARV)μNS蛋白和羊抗兔IgG分别划膜作为检测线(T线)和质控线(C线),优化试验条件,制备检测禽呼肠孤病毒血清抗体的胶体金试纸条,并检验试纸条的特异性、敏感性、重复性和稳定性。通过对90份临床血清样品进行检测并与商品化ARV抗体ELISA检测试剂盒进行比较,以评价该试纸条的准确性和实用性。结果显示,该试纸条能够特异性检出禽呼肠孤病毒阳性血清抗体,且对其他常见禽病病原血清抗体检测为阴性。敏感性试验表明禽呼肠孤病毒血清抗体的检测限为1∶160,不同批次的试纸条检测同一份样品结果均一致。试纸条保存4周后,其检测结果与新鲜制备的试纸条检测结果一致。研制的试纸条与商品化ELISA试剂盒检测临床血清样品的符合率为94.4%。表明研制的胶体金试纸条能特异准确地检测ARV抗体,且操作简便、快捷,灵敏度较高,重复性和稳定性良好,可应用于禽呼肠孤病毒抗体的检测。To establish a rapid and specific method for detecting antibodies against avian reovirus(ARV),the colloidal gold labeled rabbit anti-chicken IgG was used to prepare the gold-labeled pad.The ARVμNS protein and sheep anti-rabbit IgG were immobilized on the membrane as the test line(T line)and control line(C line),respectively.The experimental conditions were optimized to prepare colloidal gold test strips for detecting ARV serum antibodies,and the specificity,sensitivity,repeatability,and stability of the test strips were evaluated.The accuracy and practicality of the test strips were assessed by testing 90 clinical serum samples,and compared with commercialized ARV antibody ELISA kits.The results showed that the test strips could specifically detect positive serum antibodies against ARV,while tests for other common avian pathogen serum antibodies were negative.The sensitivity limit for detecting ARV serum antibodies was 1:160,and the results were consistent across different batches of test strips when testing the same sample.After storing the test strips for four weeks,their detection results were consistent with those of freshly prepared test strips.The concordance rate of the test strips with commercialized ELISA kits for clinical serum samples was 94.4%.The results indicate that the colloidal gold test strips prepared in this study can specifically and accurately detect ARV antibodies,and they are easy to operate,fast,highly sensitive,repeatable and stable,making them suitable for clinical detection of ARV antibodies.

关 键 词：禽呼肠孤病毒 μNS蛋白 胶体金试纸条 抗体检测 

分 类 号：S855.3[农业科学—临床兽医学]