ROLLING CIRCLE AMPLIFICATION ANALYSIS OF CERVICAL CELLS

人类乳头瘤病毒(HPV)的一部分与宫颈癌有关。宫颈癌相关HPV类型的确定可作为宫颈标本巴氏涂片筛查的辅助手段。这种分析可以减少浸润性宫颈癌的发生率，同时减少与巴氏涂片筛查和反射性组织学分析相关的人力和经济成本。HPV分析可以区分一过性感染和稀有细胞的存在，前者缓解而不发展为高级别宫颈病变，后者已被HPV生物学改变，并正在从局限性的上皮内病变进展为浸润性癌。该项目将开发一种方法，使用滚环扩增来分析宫颈样本中的单个细胞，并建立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方法来检测传统细胞学遗漏的罕见的HPV转化细胞，并验证RCA方案，以针对HPV基因表达的特征进行细胞分子分析，这些特征代表肿瘤进展的风险，而不是一过性感染。拟议的商业应用：该项目的成功完成将提供两种类型的产品。第一种方法可以作为巴氏涂片的辅助检查，在细胞学检查遗漏的罕见异常宫颈细胞的“二次检查”中提供提示。最终的产品将通过提供分子信息来增强或取代每年一次的巴氏涂片，预测早期宫颈癌损害，而这些信息对于训练有素的细胞技术学家或病理学家来说是不可见的。

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.