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Journal of Clinical Microbiology
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Mycobacteriology and Aerobic Actinomycetes

Mycobacterium malmoense-Specific Nested PCR Based on a Conserved Sequence Detected in Random Amplified Polymorphic DNA Fingerprints

Juha Kauppinen, Rauno Mäntyjärvi, Marja-Leena Katila
Juha Kauppinen
Department of Clinical Microbiology, University of Kuopio, FIN-70211 Kuopio, and
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Rauno Mäntyjärvi
Department of Clinical Microbiology, University of Kuopio, FIN-70211 Kuopio, and
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Marja-Leena Katila
Department of Clinical Microbiology, Kuopio University Hospital, FIN-70210 Kuopio, Finland
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DOI: 10.1128/JCM.37.5.1454-1458.1999
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ABSTRACT

Mycobacterium malmoense is an opportunistic human pathogen of increasing clinical importance. Since it is difficult to detect and identify the organism by conventional techniques, it was decided to seek a nucleic acid amplification method specific forM. malmoense. The method was based on detection of a conserved band in random amplified polymorphic DNA (RAPD) fingerprints of 45 M. malmoense strains. This band was a 1,046-bp product which was proven to be M. malmoense specific in dot blot hybridization analysis with a panel of mycobacterial strains belonging to 39 other species. The fragment was sequenced, and oligonucleotide primers were synthesized to evaluate the specificity of the PCR. Two primer pairs were found to be specific and sensitive in the nested PCR that was developed. All 49 M. malmoensestrains analyzed produced a PCR product of the expected size. In contrast, no strains belonging to the other mycobacterial species tested produced amplicons with these primers under specified reaction conditions. The results of the electrophoresis were confirmed by the hybridization with the M. malmoense-specific oligonucleotide probe. This method could be applied to the analysis of clinical or environmental samples, permitting the rapid detection ofM. malmoense.

  • Copyright © 1999 American Society for Microbiology
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Mycobacterium malmoense-Specific Nested PCR Based on a Conserved Sequence Detected in Random Amplified Polymorphic DNA Fingerprints
Juha Kauppinen, Rauno Mäntyjärvi, Marja-Leena Katila
Journal of Clinical Microbiology May 1999, 37 (5) 1454-1458; DOI: 10.1128/JCM.37.5.1454-1458.1999

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Mycobacterium malmoense-Specific Nested PCR Based on a Conserved Sequence Detected in Random Amplified Polymorphic DNA Fingerprints
Juha Kauppinen, Rauno Mäntyjärvi, Marja-Leena Katila
Journal of Clinical Microbiology May 1999, 37 (5) 1454-1458; DOI: 10.1128/JCM.37.5.1454-1458.1999
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KEYWORDS

DNA Fingerprinting
mycobacterium
polymerase chain reaction
Random Amplified Polymorphic DNA Technique

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