水牛伊氏锥虫病免疫胶体金试纸条的研制及初步应用  

Preparation and Primary Application of Immunity Colloidal Gold Test Strip for Water Buffalo Trypanosomiasis

作　　者：孟盟[1] 曹池[1] 陈汉忠[1] 刘明如[1] 王华俊[1] 奉彬[1] 陈国宝[1] 

机构地区：[1]广西大学动物科学技术学院,南宁530005

出　　处：《畜牧兽医学报》2011年第7期988-993,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金资助项目(30660140)

摘　　要：为建立一种快速、简便、适应现场应用的水牛伊氏锥虫病诊断方法,采用柠檬酸三钠还原法制备胶体金颗粒,标记纯化的抗伊氏锥虫VSG抗原的OB6单克隆抗体,在硝酸纤维素膜上包被纯化的抗伊氏锥虫VSG抗原的TB7单克隆抗体和兔抗小鼠IgG,作为检测带和质控带,然后优化条件,研制成检测伊氏锥虫病的免疫胶体金试纸条。通过交叉试验表明该试纸条与衣原体、弓形虫、旋毛虫、巴贝斯焦虫和大片吸虫无交叉反应。该试纸条最低可以检出1∶3 200倍稀释的伊氏锥虫VSG抗原(浓度为3.11mg.mL-1)。应用该试纸条对230份水牛血清样本进行初步检测,同时用ELISA做平行试验,阳性符合率为88.2%。结果证明该试纸条是一种快速、灵敏、特异的水牛伊氏锥虫病的检测方法,为水牛伊氏锥虫病的现场检测诊断和预控提供了有效的方法。To establish a rapid,simple,sensitive and adapting field application assay for water buffalo Trypanosoma evansi diseases,the colloidal gold particles was prepared by the sodium citrate reduction method.Purified OB6 monoclonal antibodies of anti-Trypanosoma evansi VSG antigen was labelled with the colloidal gold particles.The detection zone and quality control zone of the nitrocellulose membrane was made by appending purified anti-Trypanosoma evansi VSG antigen TB7,monoclonal antibody and rabbit anti-mouse IgG.The immune colloidal gold test strip was developed by optimizing the experiment conditions.The crossing test showed that the strip can＇t cross react with Chlamydia,Toxoplasma gondii,Trichina,Babesia coke worms and Fasciolagigantica.The sensitivity test showed that the strip can detected in the lowest dilution of 1：3 200 Trypanosoma evansi VSG antigen（concentration 3.11 mg·mL-1）.230 clinic serum samples of water buffalo were detected by using the test strip,while a parallel experiment were conducted using ELISA,and the positive consistent rate of both was 88.2%.All of the results showed the strip is a rapid,sensitive and specific method for detecting water buffalo Trypanosoma evansi infection.This new method has provided a useful tool for diagnose,prevention and control for Trypanosoma evansi diseases in water buffaloes.

关 键 词：伊氏锥虫病 单克隆抗体 胶体金 试纸条 

分 类 号：S854.4[农业科学—临床兽医学] S852.72[农业科学—兽医学]