猴免疫缺陷病毒(SIV)双抗原夹心ELISA诊断试剂盒的研制及应用  

Development of Double-antigen Sandwich ELISA Kit for Detection Antibodies Against Simian Immunodeficiency Virus

作　　者：季芳[1] 刘晓明[1] 饶军华[1] 

机构地区：[1]华南濒危动物研究所灵长类研究开发中心,广州510555

出　　处：《实验动物科学》2008年第3期26-29,共4页Laboratory Animal Science

基　　金：广州市科技攻关项目(2005Z3-E4061)

摘　　要：目的研制灵敏、特异的检测血清中猴免疫缺陷病毒(Simian Immunodeficiency Virus,SIV)的双抗原夹心ELISA(dsELISA)诊断试剂盒并进行初步应用。方法采用原核表达系统表达并纯化的人类免疫缺陷病毒Ⅰ型(HIV-1)包膜蛋白(gp41)作为包被用抗原,建立dsELISA诊断方法。通过优化反应条件和筛选试剂,研制ELISA诊断试剂盒,经敏感性、特异性和重复性试验考核试剂盒的质量,并与美国BIORELIANCE公司试剂盒进行比较,将结果经美国VRL验证,判断本试剂盒与美国BIORELIANCE公司试剂盒两者之间的符合率。结果该试剂盒具有良好的灵敏度、稳定性和准确性。4℃条件下能保存8个月,与其他血清无交叉反应,与美国BIORELIANCE公司试剂盒检测结果的符合率为96%。结论研制成功的实验猴SIV ELISA检测试剂盒可用于临床检测。Objective To develop a sensitive and specific double - antigen sandwich ELISA （dsELISA） kit for detection serum antibody of simian immunodeficiency Virus （SIV）. Methods The envelope protein of human immunodeficiency Virus type 1 （HIV-1） expressed in E. coli, was used as detective antigen in dsELISA kit. The dsELISA method was established by optimization reaction conditions and reagent selection. The sensitivity, specificity and reproducibility of the kit were also studied in this series A parallel detection of 400 monkey sera using dsELISA and a recognized standard kit （BioReliance kit, USA） was performed. The equivocal sera were confirmed by VRL Jab. Results The kit has good specificity and reproducibility. The kit is stable well when storing at 4℃ for 8 months. More than 96% of the results were coincident with the results detected by BioReliance kit and dsELISA kit. Conclusion The kit developed in this series has several advantages compared with commercial kit, and can be applied in SIV antibody detection and research.

关 键 词：猴免疫缺陷病毒 双抗原夹心ELISA法 试剂盒 

分 类 号：R446.61[医药卫生—诊断学]