NUCLEIC ACID BASED DETECTION AND QUANTITATION OF HIV-1 AND HIV-2

HIV-1 和 HIV-2 的核酸检测和定量

• 批准号: 3748264
• 负责人: I K HEWLETT
• 金额: --
• 依托单位: BUREAU OF HEALTH PLANNING AND RESOURCES DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3748264
• 关键词: AIDS /HIV test; HIV infections; Kaposi's sarcoma; clinical chemistry; diagnosis design /evaluation; human immunodeficiency virus 1; human immunodeficiency virus 2; human tissue; leukopenia; nucleic acid probes; plasma; polymerase chain reaction; saliva; virus DNA; virus RNA

Primer pairs have been developed that allow the detection and discrimination of HIV-1 and HIV-2 by PCR. When used singly, these primers and probes permit detection of between 1-10 copies of viral DNA. Greater than 90% sensitivity was seen in clinical samples for the HIV-2 primers and sensitivity was greater in samples from persons with CD4 counts less than 700. These primers can also be used in a co- amplification format with good sensitivity. These primers have been used to study the presence of HIV and related viruses in diseases of unknown etiology such as Idiopathic CD4+ T-Lymphocytopenia and Kaposi's Sarcoma. A sensitive method for amplification of RNA in the presence of UNG was developed. By this method, all steps of the PCR procedure are performed in a single tube a minimizing carryover contamination. A procedure for amplification of HIV DNA and RNA is being developed for in situ amplification and detection by PCR. This procedure is being modified for non-isotopic detection systems. These detection methods are being used to detect and quantitate virus in saliva and plasma and in infected cells from patients.

已经开发出了允许检测和 用聚合酶链式反应鉴别HIV-1和HIV-2单独使用时，这些 引物和探针可以检测到1-10个拷贝的病毒DNA。 临床样本对HIV-2的敏感性超过90% 在来自CD4携带者的样本中，引物和敏感性更高 不到700人。这些引物也可用于联合检测。 具有良好灵敏度的放大格式。这些底漆已经用过了 研究HIV及相关病毒在不明原因疾病中的存在 病因如特发性CD4+T淋巴细胞减少症和卡波西肉瘤。 建立了一种在UNG存在下扩增RNA的灵敏方法 发展起来的。通过该方法，执行了PCR程序的所有步骤 在单管中，最大限度地减少了残留污染。一种程序，用于 HIV DNA和RNA的原位扩增正在开发中 用聚合酶链式反应进行扩增和检测。此程序正在修改以用于 非同位素检测系统。这些检测方法正在被使用 检测和定量唾液、血浆和感染细胞中的病毒 从病人那里。