Differentiation of Mycobacterium species by analysis of the heat-shock protein 65 gene (hsp65).
The nucleotide sequences (604 bp) of partial heat-shock protein genes (hsp65) from 161 Mycobacterium strains containing 56 reference Mycobacterium species and 105 clinical isolates were determined and compared. hsp65 sequence analysis showed a higher degree of divergence between Mycobacterium species than did 16S rRNA gene analysis. Generally, the topology of the phylogenetic tree based on the hsp65 DNA sequences was similar to that of the 16S rRNA gene, thus revealing natural relationships among Mycobacterium species. When a direct sequencing protocol targeting 422 bp sequences was applied to 70 non-tuberculous mycobacterium (NTM) clinical isolates, all NTMs were clearly identified. In addition, an XhoI PCR restriction fragment length polymorphism analysis method for the differentiation of Mycobacterium tuberculosis complex from NTM strains was developed during this study. The results obtained suggest that 604 bp hsp65 sequences are useful for the phylogenetic analysis and species identification of mycobacteria.
Kim
Hong
H
Department of Microbiology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea.
Kim
Sun-Hyun
SH
Department of Microbiology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea.
Shim
Tae-Sun
TS
Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-600, Korea.
Kim
Mi-Na
MN
Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-600, Korea.
Bai
Gill-Han
GH
The Korean Institute of Tuberculosis, The Korean National Tuberculosis Association, Seoul 137-140, Korea.
Park
Young-Gil
YG
The Korean Institute of Tuberculosis, The Korean National Tuberculosis Association, Seoul 137-140, Korea.
Lee
Sueng-Hyun
SH
Department of Microbiology, College of Medicine, Konkuk University, Chungju 380-230, Korea.
Chae
Gue-Tae
GT
Institute of Hansen's Disease, The Catholic University Medical College, Seoul 137-7014, Korea.
Cha
Chang-Yong
CY
Department of Microbiology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea.
Kook
Yoon-Hoh
YH
Department of Microbiology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea.
Kim
Bum-Joon
BJ
Department of Microbiology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea.
D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D018834:Chaperonin 60; D018833:Chaperonins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D001426:Bacterial Proteins; D018834:Chaperonin 60; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; C074119:heat-shock protein 65, Mycobacterium; C034510:CTCGAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D018833:Chaperonins