Use of PCR and RFLP Analysis and Restriction Fragment Length Polymorphism Analys

聚合酶链式反应(PCR)扩增快速生长的分枝杆菌和诺卡氏菌的部分基因组，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中一种有用的技术。与根据生化测试进行常规鉴定所需的一个月或更长时间相比，在生物隔离后的几天内就可以在物种一级对这些生物体进行鉴定。此外，与生化检测相比，这些分子程序可以更准确地区分物种和亚种。我们对诺卡氏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，到目前为止还没有被识别的诺卡氏菌物种的存在；进一步表征这些生物的工作正在进行中。我们刚刚开始研究用高效液相色谱来区分这些生物的可行性。已经出版了一份手稿，描述了我们的方法，并总结了我们迄今的发现。

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. We are just beginning a study of the feasibility of distinguishing among these organisms by high-performance liquid chromatography. A manuscript describing our methodology and summarizing our findings to date has been published.