DEVELOPMENT OF METHODS FOR EPIDEMIOLOGIC TYPING OF PNEUMOCYSTIS CARINII

无法培养卡氏肺孢子虫已被证明是一个主要的 努力了解疾病流行病学的绊脚石 由这种有机体引起的。关于以下问题仍有许多问题需要解决 传播、感染的传染性以及疾病是否 易感人群新感染的结果或 免疫抑制期间潜伏感染的重新激活。这些 问题不能被完全解决，直到“打字”的方法 卡氏肺孢子虫的分离株是可用的。分子生物学研究进展 科技使研究人类的遗传联系成为可能。 通过核酸的放大，即使当有机体有 我们打算用这些方法来研究P. 卡里尼。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 对于菌株间的可变性正在进行研究。特别的 感兴趣的是核糖体RNA基因簇的区域 事实证明，这对其他生物的类似研究很有用。这些 这些区域将通过使用卡氏肺孢子虫特异性引物和 对产物进行了电泳法分析。如有必要，限制 将使用内切酶分析或序列测定来 加大歧视力度。

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.