Differentiation of clinical Mycobacterium tuberculosis complex isolates by their GyrB polymorphism

Purpose: To evaluate the reliability of the gyrB PCR-RFLP technique in differentiating clinical Mycobacterium tuberculosis complex isolates. Materials and Methods: A primer pair MTUB-f and MTUB-r for M. tuberculosis complex (MTBC) was used to differentiate 79 mycobacterial isolates by specific ampli...

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Bibliographic Details
Published in:Indian journal of medical microbiology 2010-01, Vol.28 (1), p.26-29
Main Authors: Abass, N. A, Suleiman, K. M, El Jalii, I. M
Format: Article
Language:English
Subjects:
Bacteria
Deoxyribonucleic acid
DNA
Genes
GyrB sequences
GyrB sequences, Mycobacterium tuberculosis complex, Mycobacteria other than tuberculosis
Microbiology
Mycobacteria other than tuberculosis
Mycobacterium tuberculosis
Mycobacterium tuberculosis complex
Thermal cycling
Tuberculosis
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Summary:Purpose: To evaluate the reliability of the gyrB PCR-RFLP technique in differentiating clinical Mycobacterium tuberculosis complex isolates. Materials and Methods: A primer pair MTUB-f and MTUB-r for M. tuberculosis complex (MTBC) was used to differentiate 79 mycobacterial isolates by specific amplification of the 1,020-bp fragment of the gyrB gene (gyrB-PCR1). The MTBC isolates were further differentiated using a set of specific primers MTUB-756-Gf and MTUB-1450-Cr that allowed selective amplification of the gyrB fragment specific for M. tuberculosis (gyrB-PCR2). The DNA polymorphisms in the 1,020-bp gyrB fragment for 7 M. tuberculosis strains confirmed by PCR as well as 2 reference strains; M. tuberculosis H37Rv and M. bovis BCG were analyzed with the restriction enzyme Rsa1. Results: Seventy-seven (97.5%) isolates were positive for gyrB-PCR1 and thus identified as members of M. tuberculosis complex (MTBC) and two (2.6%) isolates were negative and identified as Mycobacteria other than tuberculosis (MOTT). All the M. tuberculosis isolates showed the typical M. tuberculosis specific Rsa1 RFLP patterns (100, 360, 560-bp) while 360 and 480-bp fragments were generated from M. bovis BCG. Conclusion: The gyrB PCR-RFLP using the endonuclease Rsa1 can be used to differentiate M. tuberculosis from M. bovis in clinical isolates.
ISSN:0255-0857
1998-3646
DOI:10.4103/0255-0857.58724