双抗体夹心ELISA法检测HIV-1 p24抗原的初步建立  

Study of double-antibody sandwiched ELISA method to detect HIV-1 p24 antigen

作　　者：侯俊[1] 胡燕[1] 沈宏辉[1] 朱雷[1] 王志杰[1] 雷厉[1] 貌盼勇[1] 

机构地区：[1]解放军302医院传染病研究所病毒研究室,北京100039

出　　处：《中国艾滋病性病》2008年第3期226-228,共3页Chinese Journal of Aids & STD

摘　　要：目的建立敏感、特异的艾滋病病毒Ⅰ型(HIV-1)p24抗原的酶联免疫吸附试验(ELISA)检测方法,为试剂盒的研制奠定基础。方法用纯化的基因工程p24表达抗原免疫小鼠及兔子,获得单克隆抗体(单抗)及多克隆抗体(多抗),用单抗包被酶标板,辣根过氧化物酶标记多抗,初步建立HIV-1 p24抗原的ELISA检测方法。进一步与美国杜邦公司的HIV-1 p24抗原检测试剂同时检测88份HIV感染者血清和50份健康献血员血清中的p24抗原,确定其特异性;采用倍比稀释法的p24抗原标准品,测定试剂的灵敏度。结果两种方法检测88份HIV感染者血清和50份献血员血清中的p24抗原结果均为阴性(符合率为100%);自建ELISA法检测HIV-1 p24抗原的灵敏度为100pg/ml。结论初步建立了检测HIV-1 p24抗原的ELISA法,与国外同类试剂比较,本法的特异性和灵敏度均接近国外同类试剂,有较好的特异性和灵敏度,为进一步研制HIV-1 p24抗原ELISA检测试剂盒打下了基础。Objective To establish a sensitive and specific method for the detection of HIV-1 p24 antigen. Methods Monoclonal and polyclonal antibodies were obtained from immunity mice and rabbits using purified HIV-1 p24 antigen expressed by genetic engineering. Antibodies were conjugated with HRP. A ELISA method to detect HIV-1 p24 antigen was established. Eighty eight HIV positive sera and 50 normal sera were detected simultaneously by Dopont HIV-1 p24 antibody reagent and the ELISA method established by our laboratory to confirm its specificity. Results The results of detecting HIV-1 p24 antigen of 88 positive sera and 50 normal sera by two reagents were all negative, which showed that the coincident rate was 100 % . The sensitivity of our reagent to detect HIV-1 p24 antigen was 100pg/ml.Conclusion The ELISA methods of detecting HIV-1 p24 antigen has been preliminary established.Our reagent has good sensitivity and specificity as compared with imported ones, which has laid an important basis for developing a more sensitive diagnostic reagent kit for detecting HIV-1 p24 antigen.

关 键 词：艾滋病病毒Ⅰ型 P24抗原 酶联免疫吸附试验 

分 类 号：R512.91[医药卫生—内科学]