DEVELOPING A HYBRIDIZATION ASSAY FOR HTLV-I AND HTLV-II

已被广泛表征的人逆转录病毒（HTLV-I， HTLV-II、HIV-1和HIV-2）都具有外周血嗜性 单核细胞（PBMC）。血清学检测已经被广泛应用于 成功地识别出以前接触过这些病毒的人， 无法确定当前感染。病毒分离测定需要 患者PBMC与易感细胞系的长期共培养， 缺乏敏感性。一种新描述的DNA扩增 聚合酶链反应（PCR）技术似乎有很大的优势， 检测病毒DNA的可能性。然而，PCR需要很高的 训练有素的人员，昂贵的设备和大约三天的时间来执行一个 比色法微生物联合公司(MBA)建议开发一个简单的， 快速的杂交检测，以检测目前的感染， 逆转录病毒该方法采用塑料毛细管作为固相， 进行杂交测定。毛细管提供了一个非常大的 反应性表面相对于样品大小，也迫使关闭 接近反应物。这种环境会增加敏感性 具有较短的培养时间和温度要求。一种非同位素， 酶免疫放大检测系统用于指示 与已知DNA探针杂交。

The human retroviruses that have been extensively characterized (HTLV-I, HTLV-II, HIV-1, and HIV-2) all have a tropism for the peripheral blood mononuclear cells (PBMC). Serological assays have been used quite successfully to identify persons with prior exposure to these viruses but fail to determine a current infection. virus isolation assays require prolonged co-cultivation of patients' PBMC with a susceptible cell line and have been shown to lack sensitivity. A newly described DNA amplification technique, the polymerase chain reaction (PCR) appears to have a great potential for detection of viral DNA. The PCR however, requires highly trained personnel, expensive equipment and about three days to perform an assay. Microbiological Associates Inc. (MBA) proposes to develop a simple, rapid, hybridization assay to detect current infections by these retroviruses. The method employs a plastic capillary as a solid phase to perform the hybridization assay. The capillary provides a very large reactive surface relative to sample size and also forces the close proximity of reactants. This environment produces increased sensitivity with shorter incubation times and temperature requirements. A non-isotopic, enzyme immunoassay amplification detection system is used to indicate hybridization with a known DNA probe.