DEVELOPMENT OF METHODS FOR EPIDEMIOLOGIC TYPING OF PNEUMOCYSTIS CARINII

卡氏肺孢子虫流行病学分型方法的开发

• 批准号: 3752277
• 负责人: C P CARTWRIGHT
• 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/3752277
• 关键词: Pneumocystis carinii; bacterial RNA; bacterial genetics; electrophoresis; genetic strain; genetic techniques; genome; method development; microorganism classification; molecular genetics; nucleic acid sequence; polymerase chain reaction; restriction mapping; ribosomal RNA

The inability to culture Pneumocystis carinii has proven to be a major stumbling block in efforts to understand the epidemiology of disease caused by this organism. Much remains to be resolved concerning transmission, communicability of infection, and whether disease results from de novo infection of susceptible individuals or reactivation of latent infection during immunosuppression.. These issues cannot be addressed completely until methods for 'typing' isolates of P. carinii are available. Advancements in molecular technology make it possible to investigate the genetic relatedness of organisms by amplification of nucleic acid even when the organism has not been cultivated, and we intend to use thesE approaches to study P. carinii. The first stage of this project is currently underway, in which regions of the P. carinii genome that potentially could be sites for inter-strain variability are being investigated. Of particular interest are regions of the ribosomal RNA gene cluster that have proven useful for similar investigations in other organisms. These regions will be amplified by PCR using P. carinii-specific primers and the products analyzed electrophoretically. If necessary, restriction endonuclease analysis or sequence determination will be used to increase discriminatory power.

不能培养卡氏肺孢子虫已被证明是一个主要的 在努力了解疾病流行病学的绊脚石 是由这种生物引起的。仍有许多问题有待解决， 传播，传染性感染，以及疾病是否 由易感个体的新发感染引起，或 免疫抑制期间潜伏感染的再激活。这些 问题不能完全解决，直到“键入”的方法 卡氏肺孢子虫的分离株是可获得的。分子生物学进展 技术使研究遗传相关性成为可能， 通过核酸扩增的生物体，即使当生物体具有 我们打算用这些方法来研究P. carinii。该项目的第一阶段目前正在进行中， 卡氏肺孢子虫基因组的哪些区域可能是 正在研究菌株间变异性。特别 感兴趣的是核糖体RNA基因簇的区域， 证明对其他生物体的类似研究有用。这些 将使用卡氏肺孢子虫特异性引物通过PCR扩增区域， 对产物进行了结构分析。如有必要，限制 核酸内切酶分析或序列测定将用于 增加了辨别力。