Kirnbauer R, Hubbert N L, Wheeler C M, Becker T M, Lowy D R, Schiller J T
Laboratory of Cellular Oncology, National Cancer Institute, Bethesda, Md. 20892.
J Natl Cancer Inst. 1994 Apr 6;86(7):494-9. doi: 10.1093/jnci/86.7.494.
Previous studies have demonstrated that genital infection with high-risk types of human papillomavirus (HPV), most often HPV16, is the most significant risk factor for the development of cervical cancer. However, serologic assays that have been developed to identify high-risk HPV infection have either failed to associate serum reactivity with other indicators of HPV infection or have identified only a minority of HPV-infected individuals.
Our purpose was to determine whether a specifically developed enzyme-linked immunosorbent assay (ELISA) could detect IgG anti-HPV16 virion antibodies in the sera of women who had tested positive for genital HPV16 infection by DNA-based methods.
An ELISA was developed using newly developed HPV16 virus-like particles as antigens to detect anti-HPV16 virion IgG antibodies. These particles are comprised of HPV16 structural proteins that are self-assembled in insect cells after expression by recombinant baculoviruses. The sera of 122 women, whose HPV status had been previously evaluated by nucleic acid-based methods, were tested by this ELISA.
The sera of 59% of women (32 of 54) positive for genital HPV16 DNA by polymerase chain reaction (PCR) were positive in the ELISA assay compared with sera from women who had tested negative for HPV DNA (P < .0005). In contrast, 6% of HPV DNA-negative women (two of 31) and 9% of women positive for low-risk HPV6/11 DNA (one of 11) were ELISA positive by this criterion. The sera of women who were DNA positive for two additional high-risk HPV types were evaluated; the sera of 31% of HPV18-positive (four of 13) and 38% of HPV31-positive women (five of 13) were positive in the HPV16 particle ELISA. The sera of 75% of HPV16 DNA-positive women with severe dysplasias (12 of 16) gave positive ELISA results. The sera of 67% of women (28 of 42) who tested positive for HPV16 DNA by both PCR and the less sensitive ViraType assay tested positive in the ELISA compared with 33% of women (four of 12) who were positive by PCR but negative by ViraType (P < .05).
The majority of women with cervical HPV16 infection generate an IgG antibody response to conformationally dependent epitopes of HPV16 L1 that can be detected by ELISA.
This particular ELISA, or a similar one incorporating virus-like particles of additional HPV types, may be useful in determining the natural history of high-risk HPV infection and perhaps help to identify women at risk for developing cervical cancer.
既往研究表明，高危型人乳头瘤病毒（HPV）的生殖器感染，最常见的是HPV16，是宫颈癌发生的最重要危险因素。然而，已开发出的用于识别高危HPV感染的血清学检测方法，要么未能将血清反应性与HPV感染的其他指标相关联，要么仅识别出少数HPV感染个体。
我们的目的是确定一种专门开发的酶联免疫吸附测定（ELISA）能否检测出通过基于DNA的方法检测生殖器HPV16感染呈阳性的女性血清中的抗HPV16病毒体IgG抗体。
使用新开发的HPV16病毒样颗粒作为抗原开发一种ELISA，以检测抗HPV16病毒体IgG抗体。这些颗粒由HPV16结构蛋白组成，在重组杆状病毒表达后在昆虫细胞中自组装。用这种ELISA检测了122名女性的血清，这些女性的HPV状态先前已通过基于核酸的方法进行了评估。
通过聚合酶链反应（PCR）检测生殖器HPV16 DNA呈阳性的女性中，59%（54名中的32名）的血清在ELISA检测中呈阳性，而HPV DNA检测呈阴性的女性血清则不然（P <.0005）。相比之下，6%的HPV DNA阴性女性（31名中的2名）和9%的低危HPV6/11 DNA阳性女性（11名中的1名）根据该标准在ELISA检测中呈阳性。对另外两种高危HPV类型DNA呈阳性的女性血清进行了评估；13名HPV18阳性女性中的31%（4名）和13名HPV31阳性女性中的38%（5名）的血清在HPV16颗粒ELISA中呈阳性。16名患有严重发育异常的HPV16 DNA阳性女性中，75%（12名）的血清ELISA检测结果为阳性。通过PCR和较不敏感的ViraType检测均为HPV16 DNA阳性的女性中，67%（42名中的28名）的血清在ELISA检测中呈阳性，而通过PCR呈阳性但ViraType检测呈阴性的女性中这一比例为33%（12名中的4名）（P <.05）。
大多数宫颈HPV16感染的女性会产生针对HPV16 L1构象依赖性表位的IgG抗体反应，可通过ELISA检测到。
这种特定的ELISA，或包含其他HPV类型病毒样颗粒的类似检测方法，可能有助于确定高危HPV感染的自然史，或许有助于识别有患宫颈癌风险的女性。
