MULTIPLEX RT-PCR BASED TEST FOR HIV-1, HTLVI/II, HCV

尽管使用了高度敏感的血清学测试来筛查血液 捐献美国血液中HIV-1、HTLV-I/II、丙型肝炎病毒和乙肝病毒抗体 中枢，感染发作和抗体之间的延迟 反应会产生一个“窗口期”，在此期间血液可能 有传染性，但血清呈阴性。因此，感染病毒的风险较低 由于受污染的血液产品造成的传播仍然存在。在.期间 在这笔赠款的基础上，我们开发了一种基于多重RT-PCR的捕获方法 允许同时直接检测HTLV-I/II、HIV-1、 丙型肝炎病毒和乙肝病毒同时检测。在第二阶段的过程中，我们建议 进一步完善这种多重RT-PCR分析，并开发试剂盒格式 适用于临床实验室中血液产品的常规筛查。 该分析将在一系列表征良好的性能上进行验证 以及所有五种病毒的血清转换面板和大量 (&gt；1000)来自正常献血者的血液样本。这项测试将会 提供高度敏感、经济高效的传染性直接测量方法 血液中的病毒并有助于进一步缩小抗体阴性的“窗口” 感染和检测之间的时间间隔。

Despite the use of highly sensitive serologic tests for screening blood donations for antibodies to HIV-1, HTLV-I/II, HCV and HBV in US blood centers, the delay between the onset of infection and the antibody response creates a "window" period during which the blood may be infectious but seronegative. As a result, a low risk of virus transmission due to contaminated blood products still exists. During PHASEI of this grant, we have developed a Multiplex RT-PCR based capture assay which permits the simultaneous direct detection of HTLV-I/II, HIV-1, HCV and HBV in a single assay. In the course of PHASE II, we propose to further refine this Multiplex RT-PCR assay, and to develop a kit format useful for routine screening of blood products in a clinical lab setting. The assay will be validated on a number of well characterized performance and seroconversion panels for all five viruses and on large numbers (>1,000) of blood samples from normal blood donors. This test will provide a highly sensitive, cost-effective direct measure of infectious virus in blood and help further reduce the antibody-negative "window" period between infectivity and detection.