The development of serological test detecting antibodies against oncogenic human papillomaviruses by ELISA

用双抗体夹心法检测了100例宫颈病变、26例未确定意义的非典型细胞(ASCUS)和14例细胞学正常的HPVDNA患者的HPV16型病毒样颗粒(VLP)免疫球蛋白A和G(IgA和Ig G)，并以130例无HPVDNA的细胞学正常妇女为对照。用聚合酶链式反应方法检测宫颈标本中HPVDNA的存在。HPV16DNA阳性的正常妇女每隔4~7个月随访16~24个月。在HPV16DNA反复阳性的妇女中，经常检测到针对HPV16-VLP的IGA和Ig G抗体，这表明HPV的持续感染对有效的抗体应答至关重要。与免疫球蛋白反应相比，免疫球蛋白A反应出现得更早，持续时间更长。HPV16、31/33/35、58型和未知型HPVDNA携带者的IgA和Ig G抗体阳性率均显著高于对照组(P均<0.05)。在HPV18/45-DNA阳性组中未检测到明显的IgA和Ig G阳性。HPV31/33/35、58与HPV16相近，而HPV18/45在抗原性上与HPV16不同。宫颈正常、HPV DNA阳性、ASCUS阳性、HSIL阳性和宫颈癌阳性的患者中，抗HPV16-VLP的IGA和Ig G抗体阳性率明显高于对照组。在HSIL和宫颈癌患者中，高的血清学反应依赖于HPV16感染，尽管在ASCUS和LSIL等较轻微的变化中，HPV类型与血清学反应之间的关联并不明显。抗体阳性反映了持续的病毒感染，这可能增加宫颈恶性进展的风险。因此，使用HPV16-VLP的血清学检测可能作为补充宫颈细胞学检查的一种新的诊断工具。

Immunoglobulins A and G (IgA and IgG) responses against HPV 16 virus-like particles (VLP) were tested by ELISA in 100 women with cervical lesions, 26 atypical cells of undetermined significance (ASCUS) and 14 cytologically normal women with HPV DNA.As controls, 130 age-matched cytologically normal women with no HPV DNA were selected from the population in which the cases generated. The existence of HPV DNA in cervical samples were tested by PCR-based method. Normal women positive with HPV 16 DNA were followed up at 4-7 months intervals for 16-24 months. IgA and IgG antibodies against HPV 16-VLP were frequently detected in these women repeatedly positive for HPV16 DNA,suggesting that the persistent HPV infection is crucial for effective antibody responses against the viruses. IgA response appears earlier and persist longer that IgG response. Women with HPV DNA of types 16,31/33/35,58 and unknown types showed significantly higher seropostivity for both IgA and IgG antibodies than the controls (P<0.05 for both). No significant seropositivity for IgA nor IgG was detected in HPV 18/45-DNA positive group. HPV 31/33/35,58 appear to be close types to HPV 16, whereas HPV 18/45 to be distinct to HPV 16 in antigenicity. IgA and IgG responses against HPV 16-VLP were more frequently observed in women with normal cervices with HPV DNA,ASCUS,HSIL and cervical cancer than in the controls. High serological responses were dependent on HPV 16-infection in HSIL and cervical cancer cases, although the association between serological responses and HPV types were not apparent in the more beign changes such as ASCUS and LSIL.Antibody positive reflects persistent viral infection that may increase the risk for malignant progression of the cervix. Thus this serological assay using HPV 16-VLP may be useful as a new diagnostic tool supplementing cervical cytological tests.