Rapid Strip Test for Cervical Cancer via HPV-E6 Detection

描述(申请人提供)：宫颈癌是一个全球性的健康问题，每年导致超过20万人死亡，是确诊后存活率最低的疾病之一。使用巴氏涂片及早发现宫颈异常，大大减少了工业化国家的死亡人数，但即使在美国，每年也有1.2万例新病例和4000例死亡。迫切需要一种能够在护理时使用的快速、准确和低成本的诊断测试。人乳头瘤病毒(HPV)感染是。与99.7%以上的宫颈癌相关。针对检测HPV DNA的新测试可以识别感染高危HPV病毒类型的样本，但越来越多的证据表明，人体可以在这些感染进展为癌症之前清除其中90%以上，从而可能在筛查中造成大量假阳性。最近的研究表明，HPV-E6癌蛋白的表达是维持癌症状态所必需的，而E6蛋白的下调会导致细胞凋亡。因此，E6蛋白是一个有吸引力的诊断靶点，因为它与癌症表型高度相关，而不是与癌症的潜在可能性相关。Arbor Vita开发了一种快速免疫分析方法，用于检测人类细胞中的E6蛋白，使用的是一种PDZ结构域蛋白，能够结合所有高危HPV-E6蛋白和针对E6亚型的单抗。目前的检测方法可以检测到60000个宫颈癌细胞。Arbor Vita已经与Path建立了合作关系，以开发适合用作护理点诊断的侧向流动版本的该检测方法。这项建议的目的是1)确定最佳的E6检测试剂，用于侧向流动分析模式，能够检测到最流行的7种高危HPV型(在&gt；85%的宫颈癌中发现)，2)确定一种检测系统，该系统能够检测到对应于~10000个宫颈癌细胞的E6，这适合于预测宫颈癌3)确定最佳的样本处理条件，以及4)开发能够从宫颈样本中前7种类型中的任何一种中检测E6蛋白的原型。在第二阶段SBIR中，我们将在大量临床样本上测试原型疗效，并在必要时优化以供FDA提交。

DESCRIPTION (provided by applicant): Cervical cancer is a global health problem that continues to result in over 200,000 deaths a year and has one of the worst survival rates following diagnosis. Early detection of cervical abnormalities using the Pap smear has significantly reduced the number of deaths in industrialized nations, but even in the United States 12,000 new cases and 4,000 deaths are occurring each year. There is a critical need for a rapid, accurate and low cost diagnostic test that can be used at the point of care. Human papillomavirus (HPV) infection is. correlated with greater than 99.7% of cervical cancers. New tests directed at detection of HPV DNA can identify samples infected with high-risk HPV virus types, but a growing body of evidence suggests that the body can clear over 90% of these infections before they progress to cancer, thus potentially creating a large number of false positives in screening. Recent studies suggest that the HPV-E6 onco protein expression is required for maintenance of the cancer state, and that down regulation of E6 protein leads to apoptosis. Thus, the E6 protein is an attractive diagnostic target because it is highly correlated to the cancerous phenotype, not the potential for cancer. Arbor Vita has developed a rapid immunoassay for detection of E6 protein from human cells using a PDZ domain protein able to bind all high-risk HPV-E6 proteins and monoclonal antibodies against E6 subtypes. The current assay can detect 60000 cervical cancer cells. Arbor Vita has established a collaboration with PATH to develop a lateral flow version of this assay suitable for use as a point of care diagnostic. The AIMS of this proposal are to 1) identify the optimal E6 detection reagents for a lateral flow assay format capable of detecting the most prevalent 7 high-risk HPV types (found in > 85% Cervical Cancer), 2) identify a detection system that brings the assay sensitivity to detection of E6 corresponding to ~ 10000 cervical cancer cells, which is appropriate for prediction for cervical cancer 3) determine optimal conditions for sample processing, and 4) develop a prototype that is able to detect E6 protein from any of the top 7 types in a cervical sample. In a Phase II SBIR, we would test prototype efficacy on a large number of clinical samples and optimize, if necessary, for FDA submission.