Frequency of Non-tuberculosis Mycobacterium by PCR Restriction Fragment Length Polymorphism Analysis (PRA) Method

Atypical Mycobacterium cause various infections and some of them lead to Tuberculosis-like disease. Treatment of atypical mycobacterium is different from tuberculosis so identification of mycobacterium species is important for better control of tuberculosis. PRA is a more rapid and accurate method in comparison with phenotypic ones. During the present study using 3 restriction enzymes for digestion of 644 bp PCR product of hsp65 gene, identification of 50 mycobacterium isolates were accomplished. All of the 50 different atypical mycobacterium isolates from patients referred to the Pasteur Institute of Iran over 89-90 years were tested for PRA. A 644 bp fragment of hsp65 gene was amplified by PCR. Subsequently, PCR products were digested with AvaII, HphI and HpaII enzymes. Digested fragments were compared with standard algorithm and identified with GelcomparII software. Forty nine of 50 atypical Mycobacteria were identified in to 13 groups including 15 M. fortuitum, 12 M. simiae، 6 M. kansasii، 3 M. szulgai, 2 M. triviale، 2. M. gordonae، 2 M. aichiense،, 2 M. gallinarum. 1 M. hassiacum،1 M. malmoense،1 M. aurum، 1 M. marinum،1 M. abscessus and one unknown species. The results showed PRA using AvaII، HphI وHpaII is a simple, fast and accurate for identification of atypical mycobacteral isolates into species or sub species level. Rapid and exact identification of atypical mycobacterium from Mycobacterium tuberculosis is essential for effectiveness of TB surveillance programs.