Pulsed-Field Gel Electrophoresis Study of Mycobacterium abscessusIsolates Previously Affected by DNA Degradation
In: Journal of Clinical Microbiology, Jg. 42 (2004-12-01), Heft 12, S. 5582-5587
Online
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Zugriff:
ABSTRACTDNA degradation (which results in a smear pattern) occurs with almost 50% of Mycobacterium abscessusstrains during pulsed-field gel electrophoresis (PFGE). We assessed the potential benefit of using thiourea-containing buffer with M. abscessusby studying 69 isolates not previously typeable by PFGE (i.e., those with a smear pattern). Random (epidemiologically unrelated) isolates that were typeable (no smear pattern) were included as controls. Genomic DNA was digested with DraI, XbaI, and AseI. PFGE gels were run in regular gel buffer with and without 100 µM thiourea. All 69 isolates that generated smear patterns had clear band profiles when the thiourea buffer was used. These isolates were divided into only 30 patterns with DraI, 20 patterns with XbaI, and 20 patterns with AseI. The molecular profiles were all closely or possibly related, and the differences between the isolates ranged from zero to six bands. By multilocus enzyme electrophoresis (MEE), 45 of 53 smear isolates (85%) belonged to two closely related electrophoretic types. These isolates contained at least one enzyme allele seen almost exclusively in this group. Isolates without smear patterns were unaffected by thiourea and produced unrelated PFGE profiles, as well as multiple MEE types. The hsp65and 16S rRNA gene sequences of the isolates with smear patterns were identical to those of M. abscessustype strain ATCC 19977, which had a nonsmear pattern, suggesting that this clone is a subgroup within M. abscessus. This demonstrates that the inability to type M. abscessusby PFGE is associated with a single clone of organisms.
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Pulsed-Field Gel Electrophoresis Study of Mycobacterium abscessusIsolates Previously Affected by DNA Degradation
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Autor/in / Beteiligte Person: | Zhang, Yansheng ; Yakrus, Mitchell A. ; Graviss, Edward A. ; Williams-Bouyer, Natalie ; Turenne, Christine ; Kabani, Amin ; Wallace, Richard J. |
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Zeitschrift: | Journal of Clinical Microbiology, Jg. 42 (2004-12-01), Heft 12, S. 5582-5587 |
Veröffentlichung: | 2004 |
Medientyp: | serialPeriodical |
ISSN: | 0095-1137 (print) ; 1098-660X (print) |
DOI: | 10.1128/JCM.42.12.5582-5587.2004 |
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