DEVELOPMENT OF UNIVERSAL PCR ASSAY FOR RETROVIRUSES

逆转录病毒在自然界中普遍存在，长期以来被认为是 在动物中引起病原体的传染性癌症。 只是在最后一次 然而，十年来，致病性人类逆转录病毒被发现。 的 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 免疫缺陷病毒I型和II型（HIV 1和HIV 2）是病因 艾滋病的代理人-现代瘟疫。很明显，这些和其他的 未鉴定的逆转录病毒对公共健康构成了相当大的威胁 因为它们在人群中的流行率似乎在增加， 与各种疾病的联系。特别令人关切的是 它们是通过受污染的血液和血液制品传播的， 需要实施筛选程序以确保安全 的血液供应。分子生物学的最新进展，如 聚合酶链反应（PCR），有助于体外扩增 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;第一个是开发一种PCR检测方法， 基因序列在所有已知的，以及未经鉴定的人类逆转录病毒。 这是可行的，因为PCR引物靶向高度保守的 第二个主要目标是自动化整个pol基因的区域， PCR检测逆转录病毒，包括非放射性检测 特异性扩增的pol基因片段长期目标是 完善的分析，以点，它可以在血库中使用， 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.