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PCR amplification and high-resolution melting curve analysis as a rapid diagnostic method for genotyping members of the Mycobacterium avium–intracellulare complex
Article first published online: 17 FEB 2010
DOI: 10.1111/j.1469-0691.2010.03198.x
© 2010 The Authors. Journal Compilation © 2010 European Society of Clinical Microbiology and Infectious Diseases
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How to Cite
Castellanos, E., Aranaz, A. and De Buck, J. (2010), PCR amplification and high-resolution melting curve analysis as a rapid diagnostic method for genotyping members of the Mycobacterium avium–intracellulare complex. Clinical Microbiology and Infection, 16: 1658–1662. doi: 10.1111/j.1469-0691.2010.03198.x
Publication History
- Issue published online: 17 FEB 2010
- Article first published online: 17 FEB 2010
- Original Submission: 13 November 2009; Revised Submission: 29 January 2010; Accepted: 8 February 2010 Editor: M. Drancourt Article published online: 17 February 2010
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Keywords:
- High-resolution melt;
- Mycobacterium avium subspecies;
- Mycobacterium avium ssp. hominissuis;
- Mycobacterium intracellulare;
- real-time PCR
Clin Microbiol Infect 2010; 16: 1658–1662
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.