NEW METHOD FOR DETECTING CANCER AND VIRAL INFECTED CELLS

检测癌症和病毒感染细胞的新方法

• 批准号: 6173504
• 负责人: JAN TRNOVSKY
• 金额: $ 39.33万
• 依托单位: ONE CELL SYSTEMS, INC
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-15 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6173504
• 关键词: RNA; biomedical automation; cell line; communicable disease diagnosis; cytodiagnosis; cytogenetics; deltaretrovirus; diagnosis design /evaluation; flow cytometry; fluorescent dye /probe; fluorescent in situ hybridization; human T cell lymphotropic virus type 1; human T cell lymphotropic virus type 2; human immunodeficiency virus; human tissue; in situ hybridization; method development; neoplasm /cancer diagnosis; nucleic acid quantitation /detection; nucleic acid sequence; virus cytopathogenic effect

By combining microdrop in situ hybridization and flow cytometry, Phase I studies demonstrated that specific RNA sequences present in high or lower copy numbers (which comprise only 2-4% of the total RNA), including HIV-l RNA, can be detected. The assay's high sensitivity makes it a powerful diagnostic tool. In Phase II research, the assay format will be adapted to detect sub-populations of cells infected with retroviruses for both research and clinical applications. Using clinical samples, human blood cells infected with HIV-l will be identified. Using a model human cell line, assays will also be developed to identify cells infected with HIV-2, HTLV- I, and HTLV-II. Current blood based clinical assays which detect the presence of retroviruses and human T-cell lymphotrophic viruses are not optimal. Conventional fluorescence in situ hybridization methods require fixation, which reduces cell recovery as much as 50% and makes detection of small sub-populations of infected cells difficult. Conventional assays are also plagued by low hybridization efficiency and poor reproducibility. The gel microdrop (GMD) assay overcomes current limitations, including improving cell recovery to at least 90%. PROPOSED COMMERCIAL APPLICATION: By permitting rapid detection of the presence of small amounts of virus- specific nucleic acids, the proposed assay will significantly impact disease diagnosis, monitoring, and treatment, as well as blood screening.

通过将微晶体原位杂交和流式细胞仪结合在一起，I期研究表明，可以检测到高或较低拷贝数（仅占总RNA的2-4％）的特定RNA序列，包括HIV-L RNA，包括HIV-L RNA。 该测定的高灵敏度使其成为强大的诊断工具。 在II期研究中，将适应测定格式以检测用于研究和临床应用的逆转录病毒感染的细胞的亚群。使用临床样品，将发现感染HIV-L的人体血细胞。使用模型的人类细胞系，还将开发测定，以鉴定感染HIV-2，HTLV-I和HTLV-II的细胞。 目前的基于血液的临床测定法检测逆转录病毒和人类T细胞淋巴营养病毒的存在并不最佳。常规的荧光原位杂交方法需要固定，这可以降低细胞的恢复多达50％，并发现被感染细胞的小型亚群的检测变得困难。传统的测定也受到低杂交效率和差可重现性的困扰。 凝胶微旋转（GMD）测定克服了当前的局限性，包括将细胞回收率提高到至少90％。拟议的商业应用：通过允许快速检测少量病毒 - 特异性核酸的存在，拟议的测定将显着影响疾病的诊断，监测和治疗以及血液筛查。