DEVELOPMENT OF UNIVERSAL PCR ASSAY FOR RETROVIRUSES

逆转录病毒在自然界中无处不在，长期以来一直被认为是 在动物身上引起病原体的传染性癌症。只是在最后一次 然而，在发现致病性人类逆转录病毒的十年里。这个 人类嗜T淋巴细胞病毒I型和II型(HTLV 1和HTLV 2)是 与成人T细胞白血病/淋巴瘤有因果关系，而人类 I型和II型免疫缺陷病毒(HIV1和HIV2)是病原体 艾滋病的代理人--现代的瘟疫。显然，到目前为止，这些和其他 未经鉴定的逆转录病毒是一种相当大的公共卫生威胁 因为他们在人群中的流行率似乎在增加，就像 它们与各种疾病的联系。特别令人担忧的是这样一个事实 它们通过受污染的血液和血液制品传播， 有必要实施筛查程序以确保安全 血液供应的问题。分子生物学的最新进展，如 聚合酶链式反应(PCR)，便于体外扩增 DNA，为DNA诊断创造了令人兴奋的新可能性。我们的建议 有两个主要目标；第一个目标是开发一种聚合酶链式反应分析方法来扩增POL 所有已知的以及未定性的人类逆转录病毒中的基因序列。 这是可行的，因为聚合酶链式反应针对的是高度保守的 POL基因区域；第二个主要目标是使整个 聚合酶链式反应检测逆转录病毒，包括非放射性检测 特异性扩增的Pol1基因片段的同源性。长期目标是 完善化验方法，使其可以在血库中作为 可靠、灵敏、简单、自动化的逆转录病毒污染检测 血液及相关产品。

Retroviruses are ubiquitous in nature and have long been recognized as infectious cancer causing pathogens in animals. It was only in the last decade, however, that pathogenic human retroviruses were discovered. The human T lymphotropic viruses types I and II (HTLV 1 and HTLV 2) are causally associated with adult T-cell leukemia/lymphoma, while the human immunodeficiency viruses types I and II (HIV 1 and HIV 2) are the etiologic agents of AIDS - the modern plague. Clearly, these and other as yet uncharacterized retroviruses represent a considerable public health threat because their prevalence in the population appears to be increasing, as is their association with various diseases. Of particular concern is the fact that they are transmitted by contaminated blood and blood products, necessitating the implementation of screening procedures to ensure safety of the blood supply. Recent advances in molecular biology, such as the polymerase chain reaction (PCR), which facilitates in vitro amplification of DNA, create exciting new possibilities for DNA diagnostics. Our proposal has two major goals; the first is development of a PCR assay to amplify pol gene sequences in all known, as well as uncharacterized human retroviruses. This is feasible because the PCR primers are targeted to highly conserved regions of the pol gene; the second major goal is to automate the entire PCR assay to detect retroviruses, including the non-radioactive detection of the specifically amplified pol gene fragment. The long term goal is to perfect the assay to the point where it can be used in blood banks as a reliable, sensitive, simple, automated test for retrovirus contamination of blood and related products.