Zhang Y, Rajagopalan M, Brown BA, Wallace RJ; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1997 May 4-8; 97: 572 (abstract no. U168).
University of Texas Health Center, Tyler, TX.
Genetic strain comparison is essential to better define the epidemiology of MAC infection. RAPD-PCR has ben shown to be a simple and rapid technique for genotying of numerous bacterial and a few mycobacterial species. 163 MAC cultures from 33 patients with lung disease were examined by RAPD-PCR and compared with pulsed field gel electrophoresis (PFGE) done from the same subcultures. Most MAC strains were easily identified as the same (clonal) or different by PFGE with Dral, Xbal and AseI. For RAPD-PCR four primers were selected: OPA2, OPA18 (JCM 1994; 32:1827), INS-2 and IS986-FP (JCM 1994, 32:2169). DNA was prepared by standard phenol-chloroform extraction. Strains to be compared were processed and run by PCR at the same time on the same gel. Each primer produced readily discernible patterns with 1-4 major band(s) ranging from 200 bp to over 2000 bp. Variations of one major band for any given primer was rare but did occur with RAPD-PCR with strains identical by PFGE. When at least three primers were used for each group of strains from the same patient and only the differences of major bands were counted, the results of RAPD-PCR were comparable with PFGE. This study demonstrates that RAPD-PCR can be used for MAC strain comparison, but is subject to more variabilities than PFGE.
Publication Types:
Keywords:
- Electrophoresis, Gel, Pulsed-Field
- Humans
- Mycobacterium
- Mycobacterium Infections
- Mycobacterium avium Complex
- Mycobacterium avium-intracellulare Infection
- Polymerase Chain Reaction
- Random Amplified Polymorphic DNA Technique
Other ID:
UI: 102235549
From Meeting Abstracts