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W l 1 : P B e @ q B G y B G p A A P p A B G B P p O G p u w B G B P p G w .
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p k @ r I u l A @ q P L F R - R C P y k , G B G p A
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112
y 2 : P L F R - R C P G p H B s B m J R y x o . u P 1 ) u B r B o p , 2 ) u A u P B l A o k G C B , 3 ) W l A B B G B P p G w ( @ o y u v o A q ) , 4 ) W l A A v B
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113
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114
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Comparison of PCR-RFLP and Biochemical Methods for
Detection of Human Pulmonary Tuberculosis Agents
Behnam Rafiee
1
, Ali asghar Farazi
2
, Davoud Sadeghi
3
, Sepideh Ghani
4
, Razieh Nazari
5
,
Rohulah Keshavarz
6
, Keyvan Tadayon
6
, Nader Msavari
6
1
M.Sc.,Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.
2
Assistant professor, Department of Infectious and Tropical Diseases, Arak University of Medical Sciences, Arak, Iran.
3
M.Sc., Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
4
M.Sc.,Young Researchers Club, Arak Branch, Islamic Azad University, Arak, Iran.
5
Assistant Professor, Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.
6
Assistant Professor, Razi Vaccine & Serum Research Institute, Karaj, Iran.
Abstract
Background and Objective: Tuberculosis is one of important cause of death in several countries.
Mycobacterium tuberculosis complex is consisted of homogenous slow growing mycobacteria that
their isolation and identification are difficult and time consuming. The objective of this study was to
evaluate a molecular method for differentiation of Mycobacterium tuberculosis from Mycobacterium
bovis, and at the same time, to compare the method with biochemical tests.
Material and methods: In this study 68 sputum specimens were collected from the smear positive
patients who hospitalized in health centers of Markazi province. The samples were cultivated in the
specialized cultures. Also, 10 isolates of Mycobacterium bovis that were obtained from different area
of Iran as well as 6 standard strains have been used for comparison. After performance of biochemical
tests, oxyR pseudogene was amplified by PCR and the PCR products were digested by AluI endonucle-
ase.
Results: The digestion on PCR products of Mycobacterium tuberculosis and Mycobacterium bovis
showed one and three fragment panel, respectively, on the gele. Only one isolate of human
Mycobacterium bovis strain has been identified. The results of PCR-RFLP were consistent with
biochemical tests.
Conclusion: Since PCR-RFLP is a rapid and accurate method for differentiation between
Mycobacterium bovis and other Mycobacterium tuberculosis complex and the results are important in
terms of diagnosis and treatment of tuberculosis.
Keywords: Mycobacterium tuberculosis complex, PCR-RFLP, oxyR pseudogene
Journal of Microbial World
Volume 3, No. 2, June 2010
Correspondence to: Nader Mosavari
Tel: +989122611438
Email:nmosavari@yahoo.com
Journal of Microbial World 2010 3(2): 110-115
115