同时检测HIV抗体及p24抗原快速诊断试剂的研制

目的：研制可同时检测HIV－1、HIV－2抗体及p24抗原的胶体金快速诊断试剂。方法：利用重组杆状病毒－昆虫细胞系统进行HIV－1 gp41及HIV－2 gp36抗原的高效表达，以免疫亲和层析法纯化抗原。抗－HIV p24单克隆抗体杂交瘤细胞株，并制备抗－HIV p24单克隆抗体。以硝酸基纤维膜为载体，以纯化的HIV－1 gp41、HIV－2 gp36抗原及抗p24抗体点膜，20nm胶体金颗粒／抗人IgG和抗－HIV p24单克隆抗体进行标记，对33份已知HIV感染者阳性血清及6份阴性血清进行检测。结果：通过重组杆状病毒－昆虫细胞系统进行HIV－1 gp41及HIV－2 gp36抗原的表达，可获取浓度为2．0mg/L的纯化抗原。从抗p24单克隆抗体杂交瘤细胞株中培养上清液，通过葡萄球菌蛋白A免疫亲和层析柱可得到1．5mg/L的纯化抗体。利用纯化的抗原抗体进行标记，对39份已知血清进行检测，与荷兰Organon公司HIV1＋2抗体、p24抗原、ELISA诊断试剂同时检测结果进行比较，证实有较强的特异性及敏感性。结论：同时检测HIV－1、HIV－2抗体及p24抗原快速诊断试剂的问世，可为HIV感染的诊断提供一个简便、可靠、敏感的方法。

Assay for simultaneous detection of HIV p24 antigen and anti-HIV antibody

Objective To develop a rapid assay for simultaneous detection of HIV p24 antigen (Ag) and anti HIV antibody (Ab).Methods HIV 1 gp41 antigen and HIV 2 gp36 antigen were expressed by recombinant baculovirus insect system and purified by immunochromatography. p24 monoclonal antibody (mAb) was obtained from p24 hybridoma cell line.Purified antigen and mAb were dot blotted to nitrocellular membrane;20 nm colloidal gold anti human IgG ab and p24 ab complex were used for this test.Previously detected 39 sera specimens were tested in this study to compare with the result of HIV test with commercial HIV test kit.Results 20 mg/L purified gp41 Ag and gp36 Ag were obtained from recombinant baculovirus insect cell system;1.5 mg/L p24 mAb was obtained from p24 mAb hybridoma cell line. Compared the test result of 39 sera with commercial HIV test kits,consistence rate was 100%.Conclusion The rapid assay for simultaneous detection of HIV p24 antigen and anti HIV antibody provides a simple,sensitive and reliable test for HIV diagnosis.