J Clin Pathol Qual Control.
2001 Jun;23(1):201-207.
Identification of Non-tuberculosous Mycobacteria by HaeIII Restriction Enzyme Analysis
- Affiliations
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- 1Department of Clinical Pathology, Hanyang University Hospital, Seoul, Korea.
Abstract
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BACKGROUND: Mycobacteria form a heterogenous group in terms of occurrence in clinical or enviromental material, complex phenotypic and genetic data, and disease association. This study was performed to identify mycobacteria to the level of species by multiplex PCR and restriction enzyme analysis(REA).
METHODS
Eleven reference strains and 160 strains of clincal samples were used in this study. All isolates and reference strains were subcultured in Ogawa medium and identified using standard biochemical test such as niacin accumulation test and nitrate reduction test. The multiplex PCR was performed by using the primers that amplifies 65 kDa HSP gene which exist in all mycobacteria and the primers that amplifies MPB64 gene which is specific to Mycobacterium tuberculosis complex. The genus specific amplifed DNA products were digested with the restriction enzymes such as Haelll, Sau96I, and BstEII.
RESULTS
Ten cases of non-tuberculous mycobacteria were detected by multiplex PCR. The ten cases of NTM were identify as five M. intracelluare, three M. avium, two M. fortuitum subsp. peregrinum by REA using HaeIII.
CONCLUSIONS
The multiplex PCR and REA using HaeIII is the very easy and accurate method for identification of M. tuberculosis and NTM.
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