Molecular Epidemiologic study on pulmonary Mycobacterium kansasii disease

[目的]检测甘肃分枝杆菌（M.kansasii）的分子流行病学。[方法]采用热休克蛋白（hsp）65- pcr -限制性内切分析（PRA）、测序（ITS、16S-23S内部转录间隔段和hsp65）、脉冲场凝胶电泳（PFGE）、限制性内切片段长度多态性（RFLP）和基因组DNA主要多态性串联重复序列（MPTR）探针和IS1652探针对2002年6月1日至8月30日在国立医院组织金基中央胸科医学中心分离的堪萨斯氏杆菌进行检测。[结果]174株分离物中170株经hsp65-PRA鉴定为ⅰ型，2株分离物为ⅱ型，各分离物分别为ⅱb型和ⅵ型。ITS与hsp65的序列分析也通过hsp65- pra发现了相同的多态性区域。MPTR和IS1652探针的RFLPs的不同模式显示出每个均匀簇的特异性。159株经PFGE鉴定为相同的共有型(A)。与此相反，15个分离株在11个I型分离株中存在显著多态性，4个分离株在II、IIb、VI型分离株中存在显著多态性。[讨论]经PFGE验证，ⅰ型几乎是优势型，具有相同的世界主要型模式，而ⅰ型则表现为多态性模式。

[Purpose] To detect molecular epidemiology in Mycobacterium kansasii (M.kansasii). [Methods] We examined the M.kansasii isolates from sputum of patients at National Hospital Organization Kinki-chuo Chest Medical Center from June 1 2002 to August 30 by Heat shock protein(hsp)65-PCR-restriction analysis (PRA), sequencing (ITS, 16S-23S internal transcribed spacer and hsp65), pulsed-field gel electrophoresis (PFGE), and restriction fragment length polymorphism (RFLP) with the major polymorphic tandem repeat (MPTR) probe and the IS1652 probe of genomic DNA. [Results] 170 strains from 174 isolates were classified as M.kansasii type I using hsp65-PRA, while 2 isolates belonged to type II and each isolate belonged type IIb and VI, respectively. Sequence analysis of the ITS and hsp65 also identified the same region of polymorphism by hsp65-PRA.The different patterns by RFLPs with MPTR and IS1652 probe were shown specific for each homogeneous cluster. 159 isolates by PFGE were recognized the same common pattern(A). In contrast, 15 isolates by PFGE were included significant polymorphism within 11 isolates of type I, and 4 isolates among type II, IIb, VI. [Discussion] We verified the M.kansasii genotype I was almost predominant type, with same pattern of major worldwide type, whereas type I were appeared to be polymorphic patterns by PFGE.