NONINVASIVE DETECTION OF ANTIBODIES AGAINST HPV

人乳头瘤病毒（HPV）是最常见的病毒性性传播疾病（STD），并与大多数肛门生殖器恶性肿瘤有关。 HPV流行病学的研究受到无法在实验室中培养病毒和缺乏动物模型的阻碍。 目前的感染“金标准”是通过PCR或杂交技术检测肛门生殖器标本中的HPV DNA。 这些技术已经注意到HPV DNA的检测与性伴侣数量的增加有关。 然而，这些技术需要进行广泛的体格检查，即骨盆检查，不适用于大规模人群流行病学调查。 由于缺乏足够的抗原来源，HPV的血清学检测并不广泛。最近，使用各种表达系统体外生产多种类型的HPV衣壳已经产生了抗原来源，以开发用于检测HPV特异性抗体的免疫测定法。 这些测定已经鉴定了与主要衣壳蛋白的类型特异性构象表位反应的人IgG应答，并且HPV衣壳抗体的存在与HPV DNA和HPV相关疾病的检测相关。这些血清检测现在可以用于筛查更大的人群，这些人群具有HPV相关疾病的潜在风险。 虽然这代表了肛门生殖器恶性肿瘤流行病学的巨大进步，但仍有一些人群处于血清筛查困难的风险中。 青少年和其他可能有感染HPV风险的儿童可能已经暴露并具有HPV特异性抗体，但从这些个体中获得血清传统上是困难的。 因此，需要有一种方法，通过这种方法可以以侵入性较小的方式对高危个体进行HPV抗体筛查。 许多针对病毒病原体（肝炎、麻疹、腮腺炎、风疹、细小病毒和HIV）的抗体已在口腔液中检测到，并可靠地反映了这些个体的血清状态。 我们假设HPV衣壳IgG抗体可以在口腔液中检测到，这些抗体的检测将反映这些患者血清中的抗体。 我们建议同时收集900例HPV感染高危受试者的血清、唾液和口腔粘膜渗出液，并使用特征良好的捕获ELISA检测这些样本的HPV 6/11、16、18、31和45型抗体。该提案将为唾液检测的应用奠定基础，以进行大型流行病学调查，并更好地确定HPV感染和疾病的风险人群。

Human papillomavirus (HPV) is the most common viral sexually transmitted disease (STD) and has been implicated in the majority of anogenital malignancies. The study of the epidemiology of HPV is hampered by the inability to grow the virus in the laboratory and the lack of an animal model. The current "gold-standard" of infection is the detection of HPV DNA in anogenital specimens by PCR or hybridization techniques. These techniques have noted an association of the detection of HPV DNA with the increased numbers of sexual partners. However, these techniques require an extensive physical examination, i.e. pelvic, and are not applicable to large population epidemiologic surveys. Due to the lack of an adequate antigen source, serological assays for HPV are not widely available. Recently, in-vitro production of HPV capsids of numerous types using various expression systems have yielded a source of antigen to develop immunoassays for the detection of HPV specific antibodies. These assays have identified human IgG responses that react to type-specific, conformational epitopes of the major capsid protein and the presence of HPV capsid antibodies are associated with the detection of HPV DNA and HPV-related disease. These serum tests can now be used to screen larger populations at potential risk for HPV-related disease. Although this represents a tremendous advance in the epidemiology of anogenital malignancies, there are populations at risk where serum screening is difficult. Adolescents and other children who are potentially at risk for acquiring HPV infection may have been exposed and have HPV-specific antibodies, but the acquisition of serum from these individuals has traditionally been difficult. Therefore, there needs to be a means by which at-risk individuals can be screened for HPV antibodies in a less invasive manner. Numerous antibodies against viral pathogens (hepatitis, measles, mumps, rubella, parvovirus, and HIV) have been detected in oral fluids and reliably reflect the serum status of these individuals. We hypothesize that HPV capsid IgG antibodies can be detected in oral fluids and that the detection of these antibodies will be reflective of that in serum of these patients. We propose to simultaneously collect serum, expectorated saliva and oral mucosa transudate from 900 subjects who are at high risk for HPV infection and test these samples for antibodies to HPV types 6/11, 16,18, 31, and 45 using a well-characterized capture ELISA. This proposal will lay the groundwork for the application of a saliva test to perform large epidemiological surveys and better define the populations at risk for HPV infection and disease.