DISCRETE ASSAY FOR INTEGRATED HPV DNA IN CERVICAL CELLS

宫颈细胞中整合 HPV DNA 的离散检测

• 批准号: 6439149
• 负责人: PHILLIP T MOEN
• 金额: $ 4.93万
• 依托单位: ONE CELL SYSTEMS, INC
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6439149
• 关键词: cervix; cervix neoplasms; communicable disease diagnosis; cytodiagnosis; diagnosis design /evaluation; human papillomavirus; in situ hybridization; tissue /cell preparation; virus DNA

Current studies indicate that development of cervical cancer is strongly linked to the DNA integration of several subtypes of the human papillomavirus (HPV). The Pap test has helped in the early detection of precursor lesions; however, results are inconclusive when low grade lesions are detected. In these cases, a solution based hybridization method can be used to screen for the presence and viral burden of common HPV subtypes associated with cervical cancer. The presence of HPV, however, is not a useful indicator of subsequent development of cervical cancer. A test which can determine the occurrence of viral integration is a better indicator of disease. Current in situ hybridization methods have limited success in distinguishing integrated virus from episomal virus. Analysis is difficult because both episomal and integrated virus often coexist, and the "diffuse" pattern indicating episomal virus often masks the "dot" pattern indicating integrated virus. By combining single cell gel microdrop (GMD) encapsulation technology, in situ hybridization and nuclear fractionation techniques, this Phase I proposal aims to develop an assay which will unambiguously distinguish integrated virus from episomal virus. Use of GMD technology will also reduce cell loss, facilitating rare cell detection. PROPOSED COMMERCIAL APPLICATIONS: The assay will permit early detection of cervical cancer and highly specific assessment of viral pathogenic mechanisms directly linked to cervical cancer, facilitating early treatment and reducing mortality rates.

当前的研究表明，宫颈癌的发展与人乳头瘤病毒（HPV）的几种亚型的DNA整合密切相关。 PAP测试有助于早期发现前体病变。但是，当检测到低级病变时，结果尚无定论。在这些情况下，一种基于溶液的杂交方法可用于筛选与宫颈癌相关的常见HPV亚型的存在和病毒负担。但是，HPV的存在并不是随后发展宫颈癌的有用指标。可以确定病毒整合发生的测试是更好的疾病指标。 当前的原位杂交方法在区分综合病毒和偶发病毒方面的成功有限。 分析很困难，因为偶发病毒和综合病毒通常共存，并且表明偶发病毒的“弥漫性”模式通常掩盖了指示综合病毒的“点”模式。 通过将单细胞凝胶微旋转（GMD）封装技术，原位杂交和核分级技术结合，该I阶段提案旨在开发一种将明确区分综合病毒和偶发病毒的测定法。 GMD技术的使用还将减少细胞损失，从而促进稀有细胞检测。拟议的商业应用：该测定法将允许对宫颈癌的早期检测以及对与宫颈癌直接相关的病毒致病机制的高度特异性评估，从而促进早期治疗并降低死亡率。