A Novel Diagnostic Assay for Oncogenic Human Papillomaviruses

描述（由申请人提供）：Arbor Vita Corporation（AVC）基于对人乳头瘤病毒（HPV）生物学的理解，开发了一种新型宫颈癌诊断方法。HPV感染是最常见的性传播疾病之一，仅在美国每年估计有550万新感染。高危（致癌）HPV类型与几乎所有宫颈癌相关。巴氏涂片和液基细胞学筛查大大降低了宫颈癌的发病率，但巴氏试验具有很高的假阴性和假阳性率，需要训练有素的细胞学家的广泛基础设施来解释结果。一个更便宜的测试，更大的可靠性和预测价值将是巨大的临床效益。已显示高风险HPV类型的病毒编码的E6和E7蛋白对于细胞转化和癌症进展是必需的，并且已知来自高风险HPV类型而非低风险HPV类型的E6蛋白结合细胞PDZ结构域。 AVC扩展了这些研究，并证明了高危HPV和E6-PDZ结合之间的完美相关性。基于这些发现，我们开发了一种新的宫颈癌诊断检测HPV E6使用PDZ蛋白捕获。在我们的SBIR I期，AVC开发了一种新型的PDZ捕获夹心ELISA检测HPV E6，可检测75%以上的高危HPV类型，并证明其对人类癌症样本的实用性。我们能够开始从细胞中定量E6，并提高灵敏度，使E6检测在一个更小的样本比通常收集在巴氏试验。在第二阶段，我们计划完成基于PDZ的宫颈癌检测原型的开发，为临床试验做准备。具体而言，我们建议：1.扩大我们的抗体检测范围，包括95%的已知高危HPV类型。2.优化E6蛋白检测的临床样本处理。3.优化PDZ/抗体夹心ELISA用于临床实验室实施。4.扩展高危E6蛋白水平与临床细胞学分期相关的研究。

DESCRIPTION (provided by applicant): Arbor Vita Corporation (AVC) has developed a novel cervical cancer diagnostic based on an understanding of the biology of human papillomavirus (HPV). HPV infection is one of the most common sexually transmitted diseases with an estimated 5.5 million new infections per year in the U.S. alone. High-risk (oncogenic) HPV types are correlated with virtually all cervical cancers. Pap smear and liquid based cytology screening has greatly reduced the incidence of cervical cancer, but the Pap test has both high false-negative and false-positive rates and requires an extensive infrastructure of trained cytologists to interpret the results. A cheaper test with greater reliability and predictive value would be of great clinical benefit. The virally-encoded E6 and E7 proteins of high-risk HPV types have been shown to be essential for cell transformation and cancer progression and E6 proteins from high-risk HPV types, but not low-risk HPV types, are known to bind cellular PDZ domains. AVC has extended those studies and demonstrated a perfect correlation between high-risk HPV and E6-PDZ binding. Based on these findings, we have developed a novel cervical cancer diagnostic assay of HPV E6 using PDZ protein capture. In our SBIR Phase I, AVC developed a novel PDZ capture sandwich ELISA test for HPV E6 that detects over 75% of high-risk HPV types and demonstrated its utility with human cancer samples. We were able to begin quantifying E6 from cells and improved sensitivity to allow E6 detection in a much smaller sample than typically collected in a Pap test. In Phase II, we propose to complete development of our prototype PDZ-based cervical cancer test in preparation for clinical trials. Specifically, we propose to: 1. Expand our antibody detection to include 95% of known high-risk HPV types. 2. Optimize clinical sample handling for E6 protein detection. 3. Optimize the PDZ/antibody sandwich ELISA for clinical laboratory implementation. 4. Extend the studies correlating high-risk E6 protein levels and clinical cytology staging.