Investigation of genetic diversity of Mycobacterium fortuitum clinical isolated by RAPD-PCR

Subject Areas : Bacteriology

Rasa Sheini Mehrabzadeh ¹ , Azardokht Khosravi ² , Abdorazagh Hashemi ³ , Hoshang Jamali ⁴

1 - MS.c., Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran.
2 - Professor, Department of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
3 - Assistant Professor, Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz, Iran
4 - Assistant Professor, Department of Immunology, Islamic Azad University, Jahrom Branch, Jahrom, Iran

Received: 2013-09-05 Accepted : 2013-11-09 Published : 2014-03-01

Keywords: RAPD-PCR, Mycobacterium fortuitum, Molecular Typing,

Abstract :

Background and Objectives: Non-tuberculosis Mycobacteria (NTM) are important due to increase in the rate of clinical outbreaks and emerging antibiotic resistance. Mycobacterium fortuitum is the most common NTMs in Iran. This study was aimed to molecular detection and genetic diversity of clinical isolates of M. foruitum in Iran using RAPD-PCR method. Materials and Methods: This cross-sectional study was carried out on 81 isolates of NTM isolated from various clinical samples. The primary identification of M. fortuitum was performed based on Acid Fast staining and routine biochemical tests. After confirmation of the isolates based on PCR, the PCR products were digested with AvaII, HphI and HpaII. The molecular typing of the M. fortuitum isolates were performed using PAPD analysis. Results: Out of 81 tested NTM, 36 strains were confirmed as M. fortuitum. Based on RAPD primers, these isolates were classified into 10 main clusters. Most of the isolates were distributed into RAPD types of 1 (9 members, 25%), 2 (8 members, 22.2%) and 6 (6 members, 16.6%). In RAPD analysis, the major fragments were 300 bp (94.4%), followed by fragment 1000 bp (66.6%). Conclusion: The results showed high discriminatory ability of RAPD–PCR method. This analysis is able to sufficiently discriminate the genotypic diversity and to prepare epidemiologic information of the M. fortuitum isolates. RAPD techniques is simple, rapid, inexpensive, and is less complicated than most of other molecular typing methods.