Microsatellite based typing of Aspergillus fumigatus: strengths, pitfalls and solutions.
Microsatellites, or short tandem repeats (STR's), are popular tools to discriminate between microbial isolates. Here, we report on the robustness of a microsatellite panel for discrimination of Aspergillus fumigatus isolates. Two major PCR artefacts (stutter peaks and minus-A peaks) can complicate correct interpretation of STR data. We investigated the effect of alterations to the various components of the PCR amplification mixtures on these PCR artefacts and on the reproducibility of this assay. Some extreme conditions led to a loss of signal, but, under all conditions where a signal was obtained, identical typing results were produced. Furthermore, pitfalls with the exchange of results between labs are discussed. These pitfalls are primarily associated with sizing of the obtained PCR fragments.
de Valk
Hanneke A
HA
Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands.
Meis
Jacques F G M
JF
Klaassen
Corné H W
CH
D001232:Aspergillus fumigatus / Q000145:classification* / Q000235:genetics*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics*; D019075:Electrophoresis, Capillary; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results
D004271:DNA, Fungal