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 ORIGINAL ARTICLE
Year : 2010  |  Volume : 28  |  Issue : 1  |  Page : 26-29

Differentiation of clinical Mycobacterium tuberculosis complex isolates by their GyrB polymorphism


1 Department of Preventive Medicine, College of Veterinary Medicine and Animal Production, Sudan University of Science and Technology. P.O. Box 204, Khartoum North, Sudan
2 Department of Microbiology, Faculty of Veterinary Medicine, University of Khartoum, Postcode: 13314, Khartoum North, Sudan
3 Department of Preventive Medicine, Faculty of Veterinary Medicine, University of Khartoum, Postcode: 13314, Khartoum North, Sudan

Correspondence Address:
K M Suleiman
Department of Microbiology, Faculty of Veterinary Medicine, University of Khartoum, Postcode: 13314, Khartoum North
Sudan
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Source of Support: German Academic Exchange Service (DAAD), Conflict of Interest: None


DOI: 10.4103/0255-0857.58724

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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.






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