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

采用ELISA法检测了100例宫颈病变妇女、26例意义不典型细胞（ASCUS）和14例细胞学正常的HPV DNA检测妇女对HPV 16病毒样颗粒（VLP）的免疫球蛋白A和G （IgA和IgG）反应。作为对照，从病例发生的人群中选择了130名年龄匹配的细胞学上没有HPV DNA的正常女性。采用基于pcr的方法检测宫颈标本中HPV DNA的存在。HPV 16 DNA阳性的正常妇女每隔4-7个月随访16-24个月。在这些HPV16 DNA反复阳性的妇女中经常检测到针对HPV 16-VLP的IgA和IgG抗体，这表明持续的HPV感染对于有效的抗体应答至关重要。IgA反应比IgG反应出现得更早，持续时间更长。HPV DNA 16型、31/33/35、58型和未知型女性IgA和IgG抗体血清阳性率均显著高于对照组（P<0.05）。HPV 18/45-DNA阳性组血清IgA和IgG均未见明显阳性。HPV 31/33/35、58似乎与HPV 16接近，而HPV 18/45在抗原性上与HPV 16不同。与对照组相比，IgA和IgG对HPV 16-VLP的反应在宫颈正常的HPV DNA、ASCUS、HSIL和宫颈癌患者中更为常见。在HSIL和宫颈癌病例中，高血清学反应依赖于HPV 16感染，尽管在ASCUS和LSIL等更严重的变化中，血清学反应与HPV类型之间的关联并不明显。抗体阳性反映持续病毒感染，可能增加子宫颈恶性进展的风险。因此，使用HPV 16-VLP的血清学检测可能是一种新的诊断工具，补充宫颈细胞学检查。

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.