Abstract
Two PCR typing methods, based on polymorphism of the insertion sequence IS6110, were compared with Mycobacterium tuberculosis strains by using a single primer complementary to the inverted repeats of IS6110. Total M. tuberculosis DNA either was amplified directly (IS6110-PCR) or was amplified following digestion and ligation (IS6110-inverse-PCR). Both PCR techniques showed a similar degree of discrimination. Because of its simplicity, IS6110-PCR was chosen to confirm that a single M. tuberculosis strain was responsible for an outbreak of tuberculosis in a secondary school. IS6110-PCR was used to study the degree of differentiation in 85 clinical M. tuberculosis isolates from BACTEC 12B broth cultures. Results were consistent with those of the standardized IS6110 restriction fragment length polymorphism (RFLP) analysis method, showing identical PCR types for identical RFLPs, although the degree of discrimination was greater by RFLP analysis. The study concludes that due to its simplicity, IS6110-PCR is a good screening method when quick differentiation between M. tuberculosis strains is needed because BACTEC cultures may be used directly.
Full Text
The Full Text of this article is available as a PDF (269.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Butler W. R., Haas W. H., Crawford J. T. Automated DNA fingerprinting analysis of Mycobacterium tuberculosis using fluorescent detection of PCR products. J Clin Microbiol. 1996 Jul;34(7):1801–1803. doi: 10.1128/jcm.34.7.1801-1803.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedman C. R., Stoeckle M. Y., Johnson W. D., Jr, Riley L. W. Double-repetitive-element PCR method for subtyping Mycobacterium tuberculosis clinical isolates. J Clin Microbiol. 1995 May;33(5):1383–1384. doi: 10.1128/jcm.33.5.1383-1384.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Groenen P. M., Bunschoten A. E., van Soolingen D., van Embden J. D. Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis; application for strain differentiation by a novel typing method. Mol Microbiol. 1993 Dec;10(5):1057–1065. doi: 10.1111/j.1365-2958.1993.tb00976.x. [DOI] [PubMed] [Google Scholar]
- Gutiérrez M., Samper S., Gavigan J. A., García Marín J. F., Martín C. Differentiation by molecular typing of Mycobacterium bovis strains causing tuberculosis in cattle and goats. J Clin Microbiol. 1995 Nov;33(11):2953–2956. doi: 10.1128/jcm.33.11.2953-2956.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haas W. H., Butler W. R., Woodley C. L., Crawford J. T. Mixed-linker polymerase chain reaction: a new method for rapid fingerprinting of isolates of the Mycobacterium tuberculosis complex. J Clin Microbiol. 1993 May;31(5):1293–1298. doi: 10.1128/jcm.31.5.1293-1298.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linton C. J., Jalal H., Leeming J. P., Millar M. R. Rapid discrimination of Mycobacterium tuberculosis strains by random amplified polymorphic DNA analysis. J Clin Microbiol. 1994 Sep;32(9):2169–2174. doi: 10.1128/jcm.32.9.2169-2174.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Otal I., Martín C., Vincent-Lévy-Frebault V., Thierry D., Gicquel B. Restriction fragment length polymorphism analysis using IS6110 as an epidemiological marker in tuberculosis. J Clin Microbiol. 1991 Jun;29(6):1252–1254. doi: 10.1128/jcm.29.6.1252-1254.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palittapongarnpim P., Chomyc S., Fanning A., Kunimoto D. DNA fingerprinting of Mycobacterium tuberculosis isolates by ligation-mediated polymerase chain reaction. Nucleic Acids Res. 1993 Feb 11;21(3):761–762. doi: 10.1093/nar/21.3.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palittapongarnpim P., Chomyc S., Fanning A., Kunimoto D. DNA fragment length polymorphism analysis of Mycobacterium tuberculosis isolates by arbitrarily primed polymerase chain reaction. J Infect Dis. 1993 Apr;167(4):975–978. doi: 10.1093/infdis/167.4.975. [DOI] [PubMed] [Google Scholar]
- Plikaytis B. B., Crawford J. T., Woodley C. L., Butler W. R., Eisenach K. D., Cave M. D., Shinnick T. M. Rapid, amplification-based fingerprinting of Mycobacterium tuberculosis. J Gen Microbiol. 1993 Jul;139(7):1537–1542. doi: 10.1099/00221287-139-7-1537. [DOI] [PubMed] [Google Scholar]
- Ross B. C., Dwyer B. Rapid, simple method for typing isolates of Mycobacterium tuberculosis by using the polymerase chain reaction. J Clin Microbiol. 1993 Feb;31(2):329–334. doi: 10.1128/jcm.31.2.329-334.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thierry D., Brisson-Noël A., Vincent-Lévy-Frébault V., Nguyen S., Guesdon J. L., Gicquel B. Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol. 1990 Dec;28(12):2668–2673. doi: 10.1128/jcm.28.12.2668-2673.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Embden J. D., Cave M. D., Crawford J. T., Dale J. W., Eisenach K. D., Gicquel B., Hermans P., Martin C., McAdam R., Shinnick T. M. Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology. J Clin Microbiol. 1993 Feb;31(2):406–409. doi: 10.1128/jcm.31.2.406-409.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Soolingen D., Qian L., de Haas P. E., Douglas J. T., Traore H., Portaels F., Qing H. Z., Enkhsaikan D., Nymadawa P., van Embden J. D. Predominance of a single genotype of Mycobacterium tuberculosis in countries of east Asia. J Clin Microbiol. 1995 Dec;33(12):3234–3238. doi: 10.1128/jcm.33.12.3234-3238.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]