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)，与大多数肛门生殖系统恶性肿瘤有关。由于无法在实验室中培养病毒，以及缺乏动物模型，HPV的流行病学研究受到阻碍。目前感染的“金标准”是通过聚合酶链式反应或杂交技术检测肛门标本中的HPVDNA。这些技术已经注意到HPV DNA的检测与性伴侣数量的增加之间的联系。然而，这些技术需要广泛的体检，即骨盆，不适用于大规模人口流行病学调查。由于缺乏足够的抗原源，HPV的血清学检测方法并不广泛。最近，使用各种表达系统体外生产多种类型的HPV衣壳，已经产生了一种抗原源，用于开发检测HPV特异性抗体的免疫分析方法。这些检测已经确定了人类免疫球蛋白对主要衣壳蛋白的类型特异性构象表位的反应，HPV衣壳抗体的存在与HPV DNA和HPV相关疾病的检测有关。这些血清检测现在可以用来筛查更多具有HPV相关疾病潜在风险的人群。虽然这代表着肛门生殖器恶性肿瘤流行病学的巨大进步，但仍有一些高危人群很难进行血清筛查。青少年和其他可能有感染HPV病毒风险的儿童可能已经接触过HPV，并具有HPV特异性抗体，但传统上很难从这些人那里获得血清。因此，需要有一种方法，使高危个体能够以一种侵入性较小的方式筛查HPV抗体。在口腔液中检测到许多针对病毒病原体(肝炎、麻疹、腮腺炎、风疹、细小病毒和艾滋病毒)的抗体，并可靠地反映了这些人的血清状况。我们假设可以在口腔液中检测到HPV衣壳抗体，并且这些抗体的检测将反映这些患者的血清中的抗体。我们建议同时收集900名HPV感染高危人群的血清、咳出的唾液和口腔黏膜漏出液，并使用一种具有良好特征的捕获ELISA法检测这些样本中的HPV6/11、16、18、31和45型抗体。这项建议将为应用唾液测试进行大规模流行病学调查奠定基础，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.