Yafteh

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i Rapiddentification of atypical mycobacterium(Non-tuberculosis mycobacterium NTM) is important because of increase of these organisms infection in recent years. As phenotypic tests are time consuming and expensive, nowadays the molecular methods are widely used for rapid identification of mycobacterium spcies. The aim of this study was molecular differentiation of atypical mycobacteria using three primers in the PCR- RFLP method and evaluation of sensitivity of primers. Materials and Methods: This study was performed on 48 atypical mycobacterium specimens separated from pulmonary tuberculosis (PTB) patients who were indentified by phenotypic tests. Drug susceptibility testing was performed by proportional method, the fragments of the 16S-23S rRNA gene spacer and hsp65 gene were amplified by PCR method. Subsequently the amplicons were digested with enzymes namely AvaII,HphI, HpaII, BstEII and HaeIII and electrophoresed on 2% agarose gel. Results: A total of 48 isolates, 8 (16.6%) had multi-drug resistant (MDR-TB), 4 (8.3%) had susceptible and 36 (75%) had non MDR ( combined resistance). 13 (27%) were rapid growing and 73% were slow growing . By rate detection of hsp65 PCR-RFLP primer was higher than other primers. Conclusion: hsp65 PCR- RFLP method was more specific and exact for differentiation of non tuberculosis mycobacterium(NTM).

Keywords: Atypical mycobacterium, Molecular differentiation, PCR-RFLP method.

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