PHASE II--RAPID DIAGNOSIS OF INFECTIOUS DISEASES

最近发现的人类 B 细胞嗜淋巴细胞病毒 (HBLV)，也称为人类疱疹病毒 6 型 (HHV-6) 被认为在多种淋巴细胞增殖中发挥作用 疾病，包括慢性单核细胞增多症样综合征、B细胞 淋巴瘤和艾滋病。 在这笔赠款的第一阶段，我们开发了 HBLV 的非放射性 DNA 探针，并已证明 其在检测淋巴细胞中的 HBLV 序列方面的有用性 组织。 伯基特氏肿瘤组织的初步检查 使用此检测的淋巴瘤和其他 B 细胞淋巴瘤患者 已经表明 HBLV 序列存在于其中许多 肿瘤，并且在其中一些肿瘤中存在相当高的水平。 同相 II，这些研究将扩展到更多的样本。 原位杂交系统将提供给其他人 研究人员并将进一步优化以允许其日常使用 在临床实验室。 DNA“三明治”检测也将进行 开发用于直接检测血液样本中的病毒。 这个 测定将利用非常简单的样品处理条件，并将 与现有的 ELISA 格式兼容。 细腻的灵敏度 （少于 10 个病毒基因组拷贝）将通过以下方式实现 将 DNA 夹心测定与聚合酶链相结合 扩音系统。 原位和 DNA 三明治 杂交检测将商业化并提供 为研究人员和临床医生提供强大的诊断工具 淋巴组织增生性疾病。

The recently discovered Human B-Cell Lymphotropic Virus (HBLV), also known as Human Herpes Virus Type 6 (HHV-6) has been implicated in playing a role in various lymphoproliferative diseases including chronic mononucleosis-like syndrome, B-cell lymphomas, and AIDS. In Phase I of this grant, we have developed a non-radioactive DNA probe for HBLV, and have demonstrated its usefulness in detecting HBLV sequences in lymphocytes and tissues. Preliminary examination of tumor tissues from Burkitt's Lymphoma and other B-cell lymphoma patients using this assay has shown that HBLV sequences are present in many of these tumors, and occur in quite high levels in some of them. In Phase II, these studies will be extended to a larger number of samples. The in situ hybridization system will be made available to other researchers and will be further optimized to permit its routine use in the clinical laboratory. A DNA "Sandwich" assay will also be developed for the direct detection of virus in blood samples. This assay will utilize very simple sample handling conditions and will be compatible with existing ELISA formats. Exquisite sensitivity (to less than 10 copies of the viral genome) will be achieved by combining the DNA sandwich assay with the Polymerase Chain Amplification system. The in situ and DNA sandwich hybridization assays will be commercialized and will provide powerful diagnostic tools to researchers and clinicians studying lymphoproliferative disorders.