NEW METHOD FOR DETECTING CANCER AND VIRAL INFECTED CELLS

通过结合微滴原位杂交和流式细胞术，I期研究证明可以检测以高或低拷贝数存在的特异性RNA序列（其仅占总RNA的2-4%），包括HIV-1 RNA。 该检测的高灵敏度使其成为一种强大的诊断工具。 在II期研究中，将对检测方法进行调整，以检测受逆转录病毒感染的细胞亚群，用于研究和临床应用。使用临床样品，将鉴定感染HIV-1的人血细胞。使用模型人细胞系，还将开发用于鉴定感染HIV-2、HTLV-I和HTLV-II的细胞的测定。 目前检测逆转录病毒和人T细胞嗜淋巴细胞病毒存在的基于血液的临床测定不是最佳的。传统的荧光原位杂交方法需要固定，这使细胞回收率降低了50%，并且难以检测感染细胞的小亚群。常规测定还受到低杂交效率和差再现性的困扰。 凝胶微滴（GMD）测定克服了当前的限制，包括将细胞回收率提高到至少90%。拟定商业应用：通过允许快速检测少量病毒特异性核酸的存在，所提出的测定将显著影响疾病诊断、监测和治疗以及血液筛查。

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.