双抗原夹心法检测抗HIV-1/2总抗体方法的建立和评价

目的 进一步提高HIv感染诊断试剂的敏感性。方法 以人工合成的HIv—1 gp41．1(sP1)、gp41．2(sP2)、gp120(sP3)、P24(sP4)和HIV—2gp36(sP5)5条多肽，采用双抗原夹心酶链免疫吸附试验(ELISA)原理，以sP1、sP3、sP4和sP5混合包被酶标板作为因相抗原，辣根过氧化物酶标记sp1、sp2、sP4和sP5多肽为标记物，建立了检测抗HIV—1／2总抗体的双抗原夹心ELISA法。结果 检测卫生部药品生物制品检定所第2代40份质控参比血清，其特异性和灵敏度均为100％，高于间接ELISA法(特异性为90％，灵敏度为65％)。检测210份其他病种患者血清均为阴性，与雅培HIVAB试剂比较检测如份健康献血员血清和88份HIv感染者血清，符合率为100％。结论 本方法特异性强、敏感性高，操作简便，适用于献血员的筛选和临床HIv感染的检测。

Establishment of a double-antigen sandwich ELISA for detecting total antibodies to human immunodeficiency virus type 1/2

Objective To describe and evaluate a double antigen sandwich ELISA for detecting human immunodeficiency virus type 1/2 (HIV 1/2) specific antibodies. Methods The peptides gp41.1(sp1),gp41.2(sp2),gp120(sp3) and p24(sp4) of HIV 1 and gp36(sp5) of HIV 2 were artificially synthesized.Then sp1, sp3, sp4 and sp5 were used as coating antigens;sp1,sp2,sp4 and sp5 labeled with HRP were used as conjugates in this sandwich ELISA. Results The specificity and sensitivity of the assay were both 100% in detecting anti HIV of 40 control sera of the second generation panel, higher than indirect ELISA (specificity 90% and sensitivity,65%, respectively). All of 210 sera from individuals with other diseases were negative for anti HIV. The consistence rate was 100% when our sandwich ELISA and Abbott HIVAB were used to detect anti HIV in 90 healthy blood donors and 88 HIV infected individuals. Conclusion The results showed that this sandwich ELISA for detection of anti HIV is specific, sensitive and convenient,and it is suitable for screening blood donors and detecting HIV infection.