2007 Volume 82 Issue 2 Pages 103-110
[Purpose] To make molecular epidemiological analysis of Mycobacterium k ansasii (Al, k ansasii) isolates.
[Methods] We examined 174 M. kansasii is olates 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 (hsp 65PRA), sequencing (ITS, 16S-23S internal transcribed spacer, and hsp 65 for discrepant case between hsp 65-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 IS 1652probe of genomic DNA.
[Results] Of the 174 M. kansasii isolates, 170 strains were classified as M. kansasii type I using hsp 65-PRA, while two isolates belonged to type II and one each isolate to type II b and VI, respectively. Although the ITS sequence of these isolates also identified the same region of polymorphism by hsp65-PRA, only type II b might be revealed atypical type II, a transitional type from typical type II to intermediate type I by hsp 65 sequence. The polymorphic patterns by RFLPs with MPTR and 1S1652 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, Ub, and VI.
[Discussion] W e 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.
