Yoshida Shiomi, Suzuki Katsuhiro, Tsuyuguchi Kazunari, Iwamoto Tomotada, Okada Masaji, Sakatani Mitsunori
Clinical Research Center, National Hospital Organization Kinki-chuo Chest Medical Center, 1180 Nagasone-cho, Kita-ku, Sakai-shi, Osaka 591-8555 Japan.
Kekkaku. 2007 Feb;82(2):103-10.
To make molecular epidemiological analysis of Mycobacterium kansasii (M. kansasii) isolates.
We examined 174 M. kansasii isolates from clinical samples of patients at National Hospital Organization Kinki-chuo Chest Medical Center from June 1, 2002 to August 31, 2005 by polymerase chain reaction (PCR) -restriction analysis (PRA) of the heat shock protein (hsp) 65 gene (hsp65-PRA), sequencing (ITS, 16S-23S internal transcribed spacer, and hsp65 for discrepant case between hsp65-PRA and ITS sequence), 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.
Of the 174 M. kansasii isolates, 170 strains were classified as M. kansasii type I using hsp65-PRA, while two isolates belonged to type II and one each isolate to type IIb and VI, respectively. Although the ITS sequence of these isolates also identified the same region of polymorphism by hsp 65-PRA, only type II b might be revealed atypical type II, a transitional type from typical type II to intermediate type I by hsp65 sequence. The polymorphic patterns by RFLPs with MPTR and IS1652 probe were shown specific for each homogeneous cluster by hsp 65-PRA. In addition, 159 isolates were recognized the same common pattern A by PFGE analysis. In contrast, the rest 15 isolates revealed significant polymorphism within 11 isolates of type I, and 4 isolates among type II, IIb, and VI.
We verified the M. kansasii genotype I was predominant, with the same pattern of major worldwide type regions, and reflected a very tight clonal structure. Type I was furthermore indicated recognition of subtypes by PFGE analysis.
对堪萨斯分枝杆菌（M. kansasii）分离株进行分子流行病学分析。
我们采用热休克蛋白（hsp）65基因的聚合酶链反应（PCR）-限制性分析（PRA）（hsp65-PRA）、测序（ITS、16S-23S内部转录间隔区以及针对hsp65-PRA和ITS序列不一致情况的hsp65）、脉冲场凝胶电泳（PFGE）以及基因组DNA的主要多态性串联重复（MPTR）探针和IS1652探针的限制性片段长度多态性（RFLP），对2002年6月1日至2005年8月31日期间在国立医院组织近畿中央胸部医疗中心的患者临床样本中的174株堪萨斯分枝杆菌分离株进行了检测。
在174株堪萨斯分枝杆菌分离株中，170株菌株通过hsp65-PRA被分类为堪萨斯分枝杆菌I型，而2株分离株属于II型，1株分离株分别属于IIb型和VI型。尽管这些分离株的ITS序列也通过hsp 65-PRA鉴定出相同的多态性区域，但只有IIb型可能被揭示为非典型II型，即从典型II型到中间I型的过渡型，通过hsp65序列。MPTR和IS1652探针的RFLP多态性模式显示对hsp 65-PRA的每个同源簇具有特异性。此外，通过PFGE分析，159株分离株呈现相同的常见模式A。相比之下，其余15株分离株在I型的11株分离株以及II型、IIb型和VI型的其中4株分离株中显示出显著的多态性。
我们证实堪萨斯分枝杆菌I型占主导地位，具有与全球主要类型区域相同的模式，并反映出非常紧密的克隆结构。此外，I型通过PFGE分析表明存在亚型。
