Extended Bibliography: |
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Ref #: |
59979 |
Author(s): |
Roth,A.;Fischer,M.;Hamid,M.E.;Michalke,S.;Ludwig,W.;Mauch,H. |
Journal: |
J Clin Microbiol |
Title: |
Differentiation of phylogenetically related slowly growing mycobacteria based on 16S-23S rRNA gene internal transcribed spacer sequences |
Volume: |
36 |
Page(s): |
139-47 |
Year: |
1998 |
Keyword(s): |
GENBANK/X92668
GENBANK/X97632
GENBANK/X97633
GENBANK/X99217
GENBANK/X99219
GENBANK/X99220
GENBANK/Y14182
GENBANK/Y14183
GENBANK/Y14184
GENBANK/Y14185
GENBANK/Y14187
GENBANK/Y14188
GENBANK/Y14189
GENBANK/Y14190
GENBANK/Y14191
GENBANK/Y14192
Base Sequence
DNA, Ribosomal/*chemistry
Molecular Sequence Data
Mycobacterium/*classification/genetics
Phylogeny
Polymerase Chain Reaction
RNA, Ribosomal, 16S/*genetics
RNA, Ribosomal, 23S/*genetics
Transcription, Genetic
|
Remarks: |
Interspecific polymorphisms of the 16S rRNA gene (rDNA) are widely used for species identification of mycobacteria. 16S rDNA sequences, however, do not vary greatly within a species, and they are either indistinguishable in some species, for example, in Mycobacterium kansasii and M. gastri, or highly similar, for example, in M. malmoense and M. szulgai. We determined 16S-23S rDNA internal transcribed spacer (ITS) sequences of 60 strains in the genus Mycobacterium representing 13 species (M. avium, M. conspicuum, M. gastri, M. genavense, M. kansasii, M. malmoense, M. marinum, M. shimoidei, M. simiae, M. szulgai, M. triplex, M. ulcerans, and M. xenopi). An alignment of these sequences together with additional sequences available in the EMBL database (for M. intracellulare, M. phlei, M. smegmatis, and M. tuberculosis) was established according to primary- and secondary-structure similarities. Comparative sequence analysis applying different treeing methods grouped the strains into species-specific clusters with low sequence divergence between strains belonging to the same species (0 to 2%). The ITS-based tree topology only partially correlated to that based on 16S rDNA, but the main branching orders were preserved, notably, the division of fast-growing from slowly growing mycobacteria, separate branching for M. simiae, M. genavense, and M. triplex, and distinct branches for M. xenopi and M. shimoidei. Comparisons of M. gastri with M. kansasii and M. malmoense with M. szulgai revealed ITS sequence similarities of 93 and 88%, respectively. M. marinum and M. ulcerans possessed identical ITS sequences. Our results show that ITS sequencing represents a supplement to 16S rRNA gene sequences for the differentiation of closely related species. Slowly growing mycobacteria show a high sequence variation in the ITS; this variation has the potential to be used for the development of probes as a rapid approach to mycobacterial identification. |
URL: |
9431937 |
|
Ref #: |
60196 |
Author(s): |
Kim,B.J.;Lee,S.H.;Lyu,M.A.;Kim,S.J.;Bai,G.H.;Chae,G.T.;Kim,E.C.;Cha,C.Y.;Kook,Y.H. |
Journal: |
J Clin Microbiol |
Title: |
Identification of mycobacterial species by comparative sequence analysis of the RNA polymerase gene (rpoB) |
Volume: |
37 |
Page(s): |
1714-20 |
Year: |
1999 |
Keyword(s): |
GENBANK/AF057449
GENBANK/AF057450
GENBANK/AF057451
GENBANK/AF057452
GENBANK/AF057453
GENBANK/AF057454
GENBANK/AF057455
GENBANK/AF057456
GENBANK/AF057457
GENBANK/AF057458
GENBANK/AF057459
GENBANK/AF057460
GENBANK/AF057461
GENBANK/AF057462
GENBANK/AF057463
GENBANK/AF057464
GENBANK/AF057465
GENBANK/AF057466
GENBANK/AF057467
GENBANK/AF057468
GENBANK/AF057469
GENBANK/AF057470
GENBANK/AF057471
GENBANK/AF057472
GENBANK/AF057473
GENBANK/AF057474
GENBANK/AF057475
GENBANK/AF057476
GENBANK/AF057477
GENBANK/AF057478
etc.
Amino Acid Sequence
DNA-Directed RNA Polymerases/chemistry/*genetics
Humans
Molecular Sequence Data
Mycobacterium/*classification/enzymology/genetics
Mycobacterium Infections/microbiology
Phylogeny
Restriction Mapping
Sequence Alignment
Sequence Homology, Amino Acid
|
Remarks: |
For the differentiation and identification of mycobacterial species, the rpoB gene, encoding the beta subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 44 reference strains of mycobacteria and clinical isolates (107 strains) by PCR. The nucleotide sequences were directly determined (306 bp) and aligned by using the multiple alignment algorithm in the MegAlign package (DNASTAR) and the MEGA program. A phylogenetic tree was constructed by the neighbor-joining method. Comparative sequence analysis of rpoB DNAs provided the basis for species differentiation within the genus Mycobacterium. Slowly and rapidly growing groups of mycobacteria were clearly separated, and each mycobacterial species was differentiated as a distinct entity in the phylogenetic tree. Pathogenic Mycobacterium kansasii was easily differentiated from nonpathogenic M. gastri; this differentiation cannot be achieved by using 16S rRNA gene (rDNA) sequences. By being grouped into species-specific clusters with low-level sequence divergence among strains of the same species, all of the clinical isolates could be easily identified. These results suggest that comparative sequence analysis of amplified rpoB DNAs can be used efficiently to identify clinical isolates of mycobacteria in parallel with traditional culture methods and as a supplement to 16S rDNA gene analysis. Furthermore, in the case of M. tuberculosis, rifampin resistance can be simultaneously determined. |
URL: |
10325313 |
|
Ref #: |
75210 |
Author(s): |
Stone,B.B.;Nietupski,R.M.;Breton,G.L.;Weisburg,W.G. |
Journal: |
Int J Syst Bacteriol |
Title: |
Comparison of Mycobacterium 23S rRNA sequences by high-temperature reverse transcription and PCR |
Volume: |
45 |
Page(s): |
811-9 |
Year: |
1995 |
Keyword(s): |
GENBANK/U24502
GENBANK/U24503
GENBANK/U24504
GENBANK/U24505
GENBANK/U24506
GENBANK/U24507
GENBANK/U24508
GENBANK/U24509
GENBANK/U24510
GENBANK/U24511
GENBANK/U24512
GENBANK/U24513
GENBANK/U24514
GENBANK/U24515
GENBANK/U24516
GENBANK/U24517
GENBANK/U24518
GENBANK/U24519
GENBANK/U24520
GENBANK/U24521
GENBANK/U24522
GENBANK/U24523
GENBANK/U24524
GENBANK/U24525
GENBANK/U24526
GENBANK/U24527
GENBANK/U24528
GENBANK/U24529
GENBANK/U24530
GENBANK/U24531
Base Sequence
Molecular Sequence Data
Mycobacterium/*genetics
Phylogeny
*Polymerase Chain Reaction
RNA, Bacterial/*chemistry
RNA, Ribosomal, 16S/chemistry
RNA, Ribosomal, 23S/*chemistry
Temperature
Transcription, Genetic
|
Remarks: |
We describe a modified rRNA sequence analysis method which we used to determine the phylogenetic relationships among 58 species belonging to the genus Mycobacterium. We combined the sensitivity of the reverse transcriptase PCR for amplifying nanogram amounts of template rRNA material with the elevated extension temperatures used for the thermostable DNA polymerase from Thermus thermophilus. A 70 degrees C reverse transcription extension step permitted improved read-through of highly structured rRNA templates from members of the genus Mycobacterium, which have G+C contents of 66 to 71 mol%. The nucleic acid sequences of the amplified material were then determined by performing thermal cycle sequencing with alpha-33P-labeled primers, again with extension at 70 degrees C. Nonspecifically terminated bands were chased by using terminal deoxynucleotidyl transferase. Our method had a template requirement of nanogram amounts or less of purified RNA or 2,000 CFU of intact cells and had sufficient sensitivity so that lyophils obtained from the American Type Culture Collection could be used as source material. Sequences from a 250-nucleotide stretch of the 23S rRNA were aligned, and phylogenetic trees were evaluated by using the De Soete distance treeing algorithm and Rhodococcus bronchialis as the outgroup. Our 23S rRNA trees were compared with previously published 16S rRNA trees, including the comprehensive trees developed by the University of Illinois Ribosomal Database Project, and included 15 species not evaluated previously. Most of the groups were in general agreement and were consistent with relationships determined on the basis of biochemical characteristics, but some new relationships were also observed. |
URL: |
7547304 |
|
Ref #: |
1300 |
Author(s): |
Skerman,V.B.D.;McGowan,V.;Sneath,P.H.A.(ed) |
Journal: |
Int. J. Syst. Bacteriol. |
Title: |
Approved Lists of Bacterial Names. |
Volume: |
30 |
Page(s): |
225-420 |
Year: |
1980 |
|
Ref #: |
6111 |
Author(s): |
Marks,J.;Jenkins,P.A.;Tsukamura,M. |
Journal: |
Tubercle |
Title: |
Mycobacterium szulgai - a new pathogen. |
Volume: |
53 |
Page(s): |
210-214 |
Year: |
1972 |
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