Pcr And Rflp Analysis As Diagnostic Tests

PCR 和 Rflp 分析作为诊断测试

• 批准号: 6542074
• 负责人: Frank G Witebsky
• 金额: --
• 依托单位: CLINICAL CENTER
• 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/6542074
• 关键词: Actinomycetales; Mycobacterium; bacterial genetics; communicable disease diagnosis; diagnosis design /evaluation; diagnosis quality /standard; diagnostic tests; microorganism classification; polymerase chain reaction; rapid diagnosis; restriction fragment length polymorphism

Polymerase chain reaction (PCR) amplification of a portion of the genome of both rapidly growing Mycobacteria and Nocardia species, followed by restriction fragment length polymorphism (RFLP) analysis of the amplification products, has proven to be a useful technique in the diagnostic laboratory. Identification of these organisms at the species level can be obtained within a few days of organism isolation, as compared with the month or more required for conventional identification based on biochemical testing. In addition, these molecular procedures allow more accurate discrimination among species and subspecies than is possible with biochemical testing. Our work with two different areas of the Nocardia genome (a portion of the gene for 16S ribosomal RNA and a portion of the gene for the heat-shock protein) has suggested the existence of hitherto unrecognized Nocardia species; work is ongoing to characterize these organisms further. In addition, we have found some species of Nocardia to be human pathogens that were not previously reported to cause disease in patients in the western hemisphere, or were not known to be human pathogens at all, so far as we know. A manuscript describing our methodology has been published. We are currently writing up some of our findings on the unusual Nocardia species that we have found to be agents of disease in man.

对快速生长的分枝杆菌和诺卡菌的部分基因组进行聚合酶链反应（PCR）扩增，然后对扩增产物进行限制性片段长度多态性（RFLP）分析，已被证明是诊断实验室的一种有用技术。与基于生化测试的常规鉴定需要一个月或更长时间相比，在生物分离后几天内就可以在物种一级对这些生物进行鉴定。此外，与生物化学测试相比，这些分子程序可以更准确地区分物种和亚种。我们对诺卡菌基因组的两个不同区域（16S核糖体RNA基因的一部分和热休克蛋白基因的一部分）的研究表明，迄今为止尚未被认识的诺卡菌物种的存在；进一步鉴定这些生物的工作正在进行中。此外，据我们所知，我们发现诺卡菌的一些种类是人类病原体，这些病原体以前没有在西半球的患者中引起疾病的报道，或者根本不知道它们是人类病原体。一份描述我们方法的手稿已经出版。我们目前正在写一些关于诺卡菌的发现，我们发现诺卡菌是人类疾病的病原体。