Detection of Mycobacterium tuberculosis complex by gyrB PCR in patients with clinical suspicious of tuberculosis in Mazandaran, Iran

ABSTRACT

Introduction: Mycobacterium tuberculosis complex [MTBC] are causative agents of human and animal tuberculosis. Differentiation of MTBC members is required for appropriate treatment of individual patients and for epidemiological purposes and reduction in drug resistances. Material and Methods: 1345 patients were collected with clinical suspicions of tuberculosis who referred to the Health Care Center of Mazandaran province from July 2010 to June 2011. The specimens were stained by the Ziehl-Neelsen staining technique and were cultured on Lowenstein-Jensen medium to detect the mycobacteria. For recognition of Mycobacterium tuberculosis complex MTUB-f and MTUB-r primer [gyrB-PCR1] were used. For differentiation of Mycobacterium tuberculosis complex members MTUB-756-Gf and MTUB- 1450Cr [gyrB-PCR2] and RFLP PCR using RsaI restriction enzymewere used. Results: Of 1345 specimens, only 65[4.83%] isolates were positive culture of which59 [90.76%] were MTBC and 6 [9.24%] identified as Mycobacteria other than tuberculosis. All of 59 isolates were M. tuberculosis. Conclusion: The gyrB-RFLP PCR and using the RsaI restriction enzyme is a rapid and easy technique to perform for differentiation of the member of M. tuberculosis complex. Key words gyrB- RFLP PCR- RsaI - Mycobacterium tuberculosis complex