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Microsatellite markers for typing Aspergillus fumigatus isolates
E Bart-Delabesse1, J F Humbert, E Delabesse
1Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, Créteil, France.
Journal of Clinical Microbiology
|August 15, 1998
Summary
Highly polymorphic microsatellite DNA markers effectively type Aspergillus fumigatus isolates. This method shows high discriminatory power, aiding epidemiological studies of invasive aspergillosis.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Aspergillus fumigatus is a significant human pathogen.
- Accurate typing of A. fumigatus isolates is crucial for epidemiological studies.
- Microsatellite DNA markers offer high polymorphism for genetic analysis.
Purpose of the Study:
- To investigate the utility of microsatellite markers for typing Aspergillus fumigatus isolates.
- To assess the discriminatory power of selected CA repeat microsatellites.
- To evaluate the suitability of these markers for epidemiological investigations.
Main Methods:
- Screening of an A. fumigatus DNA library with a (CA)10 oligonucleotide to identify microsatellite loci.
- Design of primers flanking four CA repeat microsatellites.
- Amplification of microsatellite loci using fluorophore-labeled primers.
- Analysis of PCR products via automatic sequencing and GeneScan software.
Main Results:
- Four CA microsatellite loci were identified and characterized.
- A high discriminatory power (0.994) was achieved with the four markers.
- Consistent results were obtained regardless of the DNA extraction method used.
- No distinct clustering between environmental and clinical isolates was observed.
Conclusions:
- Microsatellite markers are highly effective and suitable for typing Aspergillus fumigatus.
- These markers possess significant discriminatory power for epidemiological studies.
- The findings suggest all A. fumigatus isolates may be potentially pathogenic, warranting further investigation.