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Published on: November 10, 2015
Development of a temperature-switch PCR-based SNP typing method for Mycobacterium ulcerans
Katharina Röltgen1, Kobina Assan-Ampah, Emelia Danso
1Swiss Tropical and Public Health Institute, Molecular Immunology, Basel, Switzerland.
Plos Neglected Tropical Diseases
|November 21, 2012
Summary
This study developed a rapid temperature-switch PCR (TSP) method to genetically differentiate Mycobacterium ulcerans strains causing Buruli ulcer. This cost-effective tool enables monitoring the spread of Buruli ulcer clones in endemic regions.
Area of Science:
- Microbiology
- Genetics
- Epidemiology
Background:
- Mycobacterium ulcerans causes Buruli ulcer, a severe skin disease.
- M. ulcerans exhibits very low genetic diversity, complicating strain tracking.
- Previous SNP typing identified ten M. ulcerans haplotypes in Ghana.
Purpose of the Study:
- To develop a temperature-switch PCR (TSP) assay for differentiating M. ulcerans haplotypes.
- To enable genetic fingerprinting of M. ulcerans strains using conventional agarose gel electrophoresis.
- To facilitate genetic monitoring of Buruli ulcer clones in endemic areas.
Main Methods:
- Development and validation of temperature-switch PCR (TSP) assays.
- Utilizing 89 predefined single nucleotide polymorphism (SNP) loci.
- Conventional agarose gel-based analysis of PCR products.
Main Results:
- Successful development of TSP assays to distinguish ten M. ulcerans haplotypes.
- Validation of TSP assay accuracy for typing M. ulcerans.
- Establishment of TSP assays in a reference laboratory in Ghana.
Conclusions:
- TSP assays provide a cost-effective and rapid method for M. ulcerans genetic fingerprinting.
- This method allows for regular genetic monitoring of Buruli ulcer clones.
- Facilitates investigation into the spread of M. ulcerans in endemic countries.
