PCR Amplification and High-Resolution Melting Curve Analysis as a Rapid Diagnostic Method for Genotyping Members of the Mycobacterium avium–intracellulare Complex

Clin Microbiol Infect. 2010 Nov;16(11):1658-62. doi: 10.1111/j.1469-0691.2010.03198.x.

Abstract

Some of the members of the Mycobacterium avium–intracellulare (MAI) complex are recognized as human pathogens in both immunocompromised and immunocompetent patients. The current molecular methods that are available for genotyping the MAI complex members can be both expensive and technically demanding. In this report, we describe for the first time the application of a real-time PCR and high-resolution melt approach to differentiate between the complex members by targeting a member of the Pro- Pro-Glu gene family, MACPPE24. To this end, reference strains of the M. avium subspecies and Mycobacterium intracellulare were used to optimize the technique. Then, this real-time PCR–high-resolution melt approach was used to distinguish ten M. avium ssp. hominissuis field isolates from the M. intracellulare reference strain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Base Sequence
  • DNA Fingerprinting / methods*
  • DNA, Bacterial / genetics
  • Genotype
  • Humans
  • Molecular Sequence Data
  • Mycobacterium avium Complex / classification*
  • Mycobacterium avium Complex / genetics*
  • Mycobacterium avium Complex / isolation & purification
  • Mycobacterium avium-intracellulare Infection / microbiology*
  • Nucleic Acid Denaturation
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide*
  • Sequence Alignment

Substances

  • Bacterial Proteins
  • DNA, Bacterial