ROLLING CIRCLE AMPLIFICATION ANALYSIS OF CERVICAL CELLS

人乳头瘤病毒（HPV）的一个亚型与宫颈癌有关。确定与宫颈癌相关的HPV类型的存在可以作为宫颈样本巴氏涂片筛查的辅助手段。这种分析可以降低浸润性宫颈癌的发病率，同时降低与巴氏涂片筛查和反射组织学分析相关的人力和经济成本。HPV分析可以区分短暂感染，解决没有发展的高级别宫颈病变，并已被HPV生物学改变的罕见细胞的存在，并从局部上皮内病变进展为浸润性癌症。该项目将开发一种使用滚环扩增的方法来分析宫颈样本中的单个细胞，并确定HPV DNA和RNA序列的存在，并在第二阶段，建立其他标志物与HPV相关宫颈癌早期阶段的关系。该项目的主要目标是开发一种检测常规细胞学遗漏的罕见HPV转化细胞的方法，并验证RCA方案，用于细胞分子分析HPV基因表达的特征，这些特征具有肿瘤进展的风险，而不是短暂感染。建议的商业应用：该项目的成功完成将提供两种类型的产品。第一个可以作为一个辅助巴氏涂片检查提供一个指示时，“第二次检查”可能会发现罕见的异常宫颈细胞错过了细胞学。最终的产品将通过提供分子信息来增强或取代每年的巴氏涂片检查，预测早期宫颈癌病变，这是训练有素的细胞学技术人员或病理学家所看不到的。

A subset of human papillomavirus (HPV) types have been implicated in cervical cancer. The determination of the presence of the HPV types associated with cervical cancer may serve as adjunct to Pap smear screening of cervical samples. This analysis could reduce the incidence of invasive cervical cancer, while decreasing the human and economic costs associated with Pap smear screening and reflex histological analysis. HPV analysis could distinguish between transient infections that resolve without the development of high-grade cervical lesions, and the presence of rare cells that have been biologically altered by HPV, and are progressing from a localized intraepithelial lesion to invasive cancer. This project will develop a method using Rolling Circle Amplification to analyze individual cells within a cervical sample and establish the presence of HPV DNA and RNA sequences, and in Phase II, establish the relationship of other markers to the early stages of HPV-related cervical cancer. The majors goals of this project are to develop a means to detect rare HPV-transformed cells that are missed by conventional cytology and validate RCA protocols for molecular analyses of cells with respect to features of HPV gene expression that present a risk of neoplastic progression rather than a transient infection. PROPOSED COMMERCIAL APPLICATIONS: The successful completion of this project will provide two types of products. The first could serve as an adjunct Pap smears by providing an indication when a "second look" might reveal rare aberrant cervical cells missed by cytology. The ultimate product would enhance or replace the annual Pap smear by providing molecular information, predicting early cervical cancer lesions, that is not visible to a trained cytotechnologist or pathologist.