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，一期研究表明，可以检测到高拷贝数或低拷贝数（仅占总RNA的2-4%）的特定RNA序列，包括hiv - 1 RNA。该检测方法的高灵敏度使其成为一种强大的诊断工具。在II期研究中，该检测格式将适用于检测被逆转录病毒感染的细胞亚群，用于研究和临床应用。使用临床样本，将识别感染hiv - 1的人类血细胞。使用模型人类细胞系，还将开发检测方法来鉴定感染HIV-2、HTLV-I和HTLV- ii的细胞。目前基于血液的临床检测逆转录病毒和人类t细胞淋巴细胞病毒的存在并不是最佳的。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%，并且使检测感染细胞的小亚群变得困难。传统的检测方法也受到杂交效率低和重复性差的困扰。凝胶微滴（GMD）试验克服了目前的局限性，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，拟议的检测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.