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 approach was based on type-specific nucleotide sequences in MAP1506, a member of the PPE (proline-proline-glutamic acid) gene family.
Centro de Vigilancia Sanitaria Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Elena
E
Castellanos
Alicia
A
Aranaz
Jeroen
J
De Buck
D000818:Animals; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D044366:Transition Temperature
D004269:DNA, Bacterial