Rapid FISH Detection of HPV RNA in 3-Dimensional Skin Reconstructs

描述（由申请人提供）：人乳头瘤病毒（HPV）感染明确确定为宫颈癌的原因。然而，感染与癌症之间并不存在明显的因果关系，因为大多数感染是自限性的、良性的，并且从不进展为癌症。大量的分子研究已经发现了许多被认为是导致癌症后期发展的病毒机制，但迄今为止还没有解决为什么癌症进展需要多年才能发生，如果它发生的话。这在一定程度上阻碍了能够预测哪些患者将发展为更严重疾病的检测方法的发展。最广泛使用的HPV检测是基于溶液的DMA杂交捕获检测（HCA）。最近的几项流行病学研究清楚地表明，HCA具有很强的阴性预测价值;能够自信地说HPV阴性患者在未来几年内不会发展为宫颈癌。HCA也不具有高的阳性预测值，部分原因是无法区分正常的生产性感染和含有整合的病毒DNA的细胞，这些细胞在发育不良和宫颈癌中发现。使用一种新的探针标记方法，这个小企业创新研究项目提出开发一种灵敏的原位杂交检测方法，用于检测高危HPV病毒类型的RNA。该SBIR将使用体外皮肤重建模型作为临床HPV感染的替代模型来验证该检测方法。

DESCRIPTION (provided by applicant): Human Papillomavirus (HPV) infection is clearly established as the cause of cervical cancer. A distinct cause and effect relationship of infection to cancer does not, however, exist, since the majority of infections are self-limiting, benign, and never progress to cancer. A large body of molecular research has uncovered many of the viral mechanisms believed responsible for later development of cancer, but to date has not resolved why progression to cancer takes many years to occur, if it occurs at all. This, in part, has hindered development of assays capable of predicting which patients will progress to more severe disease. The most widely used assay for HPV is the solution-based DMA hybrid capture assay (HCA). Several recent epidemiological studies have clearly shown that HCA has a strong negative predictive value; the ability to confidently state that a HPV-negative patient will not develop cervical cancer over the next several years. HCA also does not have high positive predictive value, in part due to the inability to differentiate between normal productive infection, and cells containing integrated viral DNA, which are found in dysplasia and cervical cancer. Using a novel probe labeling method, this Small Business Innovation Research project proposes to develop a sensitive in situ hybridization assay for detecting RNAs of high-risk HPV viral types. This SBIR will validate the assay using in vitro skin reconstruct models as a surrogate model for clinical HPV infections.