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

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。

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

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 mon...