PHASE II--RAPID DIAGNOSIS OF INFECTIOUS DISEASES

最近发现的人类B细胞趋光性病毒 （HBLV），也称为人类疱疹病毒6型（HHV-6）， 在各种淋巴增生性疾病中发挥作用 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HBLV序列中的有用性， 组织中 Burkitt's肿瘤组织的初步检查 淋巴瘤和其他B细胞淋巴瘤患者使用该检测试剂盒 已经表明HBLV序列存在于许多这些 肿瘤，并且在其中一些中发生相当高的水平。 同相 第二，这些研究将扩展到更大数量的样本。 原位杂交系统将提供给其他 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”检测也将在 用于直接检测血液样本中的病毒。 这 分析将利用非常简单的样品处理条件， 与现有的ELISA格式兼容。 精致的灵敏度 (to少于10个拷贝的病毒基因组）将通过 将DNA夹心法与聚合酶链反应相结合 放大系统。 原位和DNA三明治 杂交分析将商业化，并将提供 强大的诊断工具，研究人员和临床医生研究 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.