[16S approximately 23S rDNA internal transcribed spacer sequence analysis and its application in mycobacterial identification].
OBJECTIVE
To study the applicability of the 16S approximately 23S rDNA internal transcribed spacer (ITS) sequences in mycobacterial identification.


METHODS
The 16S approximately 23S rDNA ITS sequences of 22 mycobacterial species (30 reference strains and 16 clinical isolates) were determined by PCR-direct sequencing. The sequences determined by this study and those reported in Genbank were analysed by Clustal program (MegAlign Package [Windows Version 4.01]; DNASTAR, Madison, Wis). The percentage similarity of sequence pairs was calculated and a cluster analysis dendrogram was constructed by PHYLIP package.


RESULTS
The 16S approximately 23S rDNA ITS sequences among mycobacterial species had highly polymorphism except that Mycobacterium tuberculosis complex (MTC) sequences were highly consistent. The similarity between sequence pairs was 30.4% to 86.5%. Mycobacterial species could be isolated by cluster analysis dendrogram. The results showed that sequence analysis of the 16S approximately 23S rDNA ITS can differentiate MTC from nontuberculous mycobacteria (NTM) and identify NTM at species level.


CONCLUSIONS
16S approximately 23S rDNA ITS sequences can be used as target genes in mycobacterial identification.
Li
Guoli
G
Tuberculosis Research Laboratory, 309th Hospital of PLA, Beijing 100091, China.
Zhuang
Yuhui
Y
Zhao
Ming
M
Wang
Guozhi
G
Fan
Yihu
Y
Chen
Baowen
B
Hu
Zhongyi
Z
D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S