用昆虫-杆状病毒系统表达的HPV6型L1蛋白检测尖锐湿疣患者血清中抗体

用杆状病毒表达系统表达人乳头瘤病毒6型（human papillomavirus type 6，HPV6）主要衣壳蛋白（major capsid protein，L1）作为抗原，对尖锐湿疣（condyloma acuminata，CA）患者抗体进行检测。采用昆虫杆状病毒系统表达HPV6L1蛋白，通过镍柱亲和层析法获得纯化抗原；以酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）检测30例CA、20例献血员和10例儿童血清中的HPV6 LI IgG抗体。感染重组杆状病毒的昆虫细胞经SDS-PAGE和Western blot检测，在大约55kD处有明显的外源蛋白表达条带。ELISA结果显示，CA组的血清阳性率为66．7％（20／30），献血员组的阳性率为15％（3／20），两组之间有显著性差异（P〈0．05）。22例HPV6型感染的CA患者有15例血清阳性（68．2％），6例HPV11型CA患者4例阳性（66．7％），1例混合感染者为阳性，1例HPV16型患者为阴性。女性CA患者的血清抗体阳性率高于男性（P=0．0052）。本研究建立的ELISA体系具有敏感性和针对低危型HPV感染的特异性。这不仅对于HPV血清流行病学研究是有价值的，而且对于临床诊断HPV感染可能具有一定的应用价值。

Use of Human Papillomavirus Type 6 Capsid Protein L1 to Detect Specific Antibodies in Patients with Condyloma Acuminata

To produce human papillomavirus (HPV) type 6 major capsid protein (L1) for screening the sera of condyloma acuminata (CA) patients, HPV 6 L1 protein was produced by recombinant baculovirus expression system and purified with affinity chromatography and was used as antigen to analyse by ELISA the sera of 30 CA patients, 20 blood donors and 10 children for the presence of IgG antibody. SDS-PAGE and Western blot showed recombinant baculovirus system could efficiently express HPV 6 L1 protein with good immunoreactivity.IgG antibody to HPV6 L1 protein was detected in sera from 20 of 30 (66.7%) CA patients and 3 of 20 (15%) blood donors (P<0.05). Among 30 cases of CA, 15 of 22 (68.2%) CA with HPV6 DNA, 4 of 6 (66.7%) CA with HPV11 DNA, one case of HPV6 and HPV11 co-infection were seropositive to HPV6 L1; the last case with HPV16 DNA, was HPV6 L1 IgG negative. CA male cases were less likely to be seropositive than female cases (P=0.0052). Using recombinant baculovirus expression system, the established ELISA for detection of antibody to HPV6 L1 protein is sensitive and specific for low risk HPV infection, and may be of value for not only sero-epidemiological investigation, but also clinical diagnosis.