Rapid Identification and Differentiation of Mycobacterium avium Subspecies paratuberculosis Types by Use of Real-Time PCR and High-Resolution Melt Analysis of the MAP1506 Locus

High-resolution melt (HRM) analysis can identify sequence polymorphisms by comparing the melting curves of amplicons generated by real-time PCR amplification. We describe the application of this technique to identify Mycobacterium avium subspecies paratuberculosis types I, II, and III. The HRM appro...

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Bibliographic Details
Published inJournal of Clinical Microbiology Vol. 48; no. 4; pp. 1474 - 1477
Main Authors Castellanos, Elena, Aranaz, Alicia, De Buck, Jeroen
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 01.04.2010
American Society for Microbiology (ASM)
Subjects
Animals
Clinical Veterinary Microbiology
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Mycobacterium avium
Mycobacterium avium subsp. paratuberculosis - classification
Mycobacterium avium subsp. paratuberculosis - genetics
Mycobacterium avium subsp. paratuberculosis - isolation & purification
Paratuberculosis - diagnosis
Paratuberculosis - microbiology
Polymerase Chain Reaction - methods
Transition Temperature
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Summary:High-resolution melt (HRM) analysis can identify sequence polymorphisms by comparing the melting curves of amplicons generated by real-time PCR amplification. We describe the application of this technique to identify Mycobacterium avium subspecies paratuberculosis types I, II, and III. The HRM approach was based on type-specific nucleotide sequences in MAP1506, a member of the PPE (proline-proline-glutamic acid) gene family.
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ISSN:0095-1137
1098-660X
DOI:10.1128/JCM.02484-09