Rapid Strip Test for Cervical Cancer via HPV-E6 Detection

描述（由申请人提供）：宫颈癌是一个全球性的健康问题，每年继续导致超过20万人死亡，并且是诊断后生存率最差的疾病之一。在工业化国家，使用巴氏涂片检查早期发现宫颈异常已经大大减少了死亡人数，但即使在美国，每年也有12，000例新病例和4，000例死亡。迫切需要一种快速、准确和低成本的诊断测试，可以在护理点使用。人乳头瘤病毒（HPV）感染是什么？与超过99.7%的宫颈癌相关。针对HPV DNA检测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，身体可以在这些感染进展为癌症之前清除90%以上的感染，因此可能会在筛查中产生大量假阳性。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，并且E6蛋白的下调导致细胞凋亡。因此，E6蛋白是一个有吸引力的诊断靶点，因为它与癌性表型高度相关，而不是癌症的可能性。Arbor Vita开发了一种快速免疫测定法，用于检测人体细胞中的E6蛋白，该免疫测定法使用能够结合所有高危HPV-E6蛋白的PDZ结构域蛋白和针对E6亚型的单克隆抗体。目前的检测方法可以检测60000个宫颈癌细胞。Arbor Vita与PATH建立了合作关系，以开发适用于床旁诊断的该检测试剂盒的侧流版本。本提案的目标是：1）确定能够检测最流行的7种高危HPV类型的侧流检测格式的最佳E6检测试剂（发现于> 85%的宫颈癌中），2）鉴定使测定灵敏度达到检测对应于约10000个宫颈癌细胞的E6的检测系统，其适合于预测宫颈癌3）确定样品处理的最佳条件，和4）开发能够检测宫颈样品中前7种类型中的任何一种的E6蛋白的原型。在II期SBIR中，我们将在大量临床样本上测试原型疗效，并在必要时进行优化，以提交FDA。

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.