Clin Mol Pathol. 1995 Jun;48(3):M124-32. doi: 10.1136/mp.48.3.m124.
Rapid identification of mycobacteria from AIDS patients by capillary electrophoretic profiling of amplified SOD gene.
Clinical molecular pathology
T J Bull, D C Shanson, L C Archard
Affiliations
Affiliations
- Medical Microbiology Department, Charing Cross and Westminster Medical School, London W6 8RF.
PMID: 16695992
PMCID: PMC407944 DOI: 10.1136/mp.48.3.m124
Abstract
Aim-Rapid differentiation of mycobacterial species at the genomic level.Methods-The manganese superoxide dismutase (SOD) gene (464 bp) and 16SrRNA (353 bp) from 104 isolates (18 species) of mycobacteria were amplified using polymerase chain reaction (PCR). Products were sequenced and a phenogram of SOD sequences derived. PCR products of SOD gene were digested with HaeIII, and restriction fragment profiles visualised using capillary electrophoresis.Results-Novel SOD sequences were found for M szulgai, M marinum, M phlei, M smegmatis, M chelonei, M paratuberculosis, M malmoense, M intracellulare serotype 7, M intracellulare serotype 18, and M celatum types 1, 2, and 3. Phylogenetic analysis indicated that 18 of 19 species studied had 8-29% interspecies and <6% intraspecies sequence diversity in the SOD gene. No consistent differences were detected between AIDS and non-AIDS isolates. M paratuberculosis showed a unique SOD sequence with a 1.1% (SD 0.5%) diversity from M avium. Capillary electrophoresis profiles were able to differentiate 16 of 18 species within 24 hours.Conclusions-A phenogram of SOD sequences clearly delineated all mycobacterial species and showed two distinct clusters, fast growing species, and the M avium complex (MAC). Within the MAC, M avium (five types), M intracellulare (five types), M scrofulaceum (two types), and M paratuberculosis (one type) could be demonstrated. Phylogenetic diversity of M celatum from MAC, previously suggested by 16SrRNA data, was confirmed. This simple and rapid method for DNA extraction, in conjunction with capillary electrophoresis of SOD restriction fragments, allows rapid identification of mycobacterial isolates.
References
- J Gen Microbiol. 1993 Oct;139(10):2423-9 - PubMed
- N Engl J Med. 1993 Sep 16;329(12):898-904 - PubMed
- Nucleic Acids Res. 1994 Sep 25;22(19):4002-8 - PubMed
- J Clin Microbiol. 1984 Oct;20(4):722-9 - PubMed
- Lancet. 1992 Nov 28;340(8831):1360 - PubMed
- Nucleic Acids Res. 1989 Oct 25;17(20):8378 - PubMed
- Int J Syst Bacteriol. 1993 Jul;43(3):539-48 - PubMed
- Res Microbiol. 1994 Mar-Apr;145(3):177-81 - PubMed
- J Infect Dis. 1993 Jul;168(1):112-9 - PubMed
- J Clin Microbiol. 1994 Nov;32(11):2801-12 - PubMed
- Adv Clin Chem. 1992;29:161-96 - PubMed
- Res Microbiol. 1994 Mar-Apr;145(3):243-52; discussion 252-61 - PubMed
- J Gen Microbiol. 1990 Sep;136(9):1915-20 - PubMed
- Infect Immun. 1990 Jun;58(6):1937-42 - PubMed
- FEBS Lett. 1988 Mar 14;229(2):256-60 - PubMed
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