Using Sequence Analysis to Identify Cultures Derived from Airborne Spores.
EDGEWOOD RESEARCH DEVELOPMENT AND ENGINEERING CENTER ABERDEEN PROVING GROUNDMD
Pagination or Media Count:
The purpose of this project was to evaluate rDNA sequencing in identifying airborne fungal spores. Universal fungal small subunit SSU ribosomal RNA primers were used with the polymerase chain reaction PCR to amplify the SSU rRNA gene from each isolate for sequencing. Sequences were analyzed through the ribosmal database project to identity the most closely related known rDNA sequence. Restriction fragment length polymorphisms RFLPs were obtained from three cultures to be used to detect each culture in mixed samples. Of the five stock cultures studied, one common fungus could be identified to genus with some confidence Mucor. For three cultures, sequence data yielded closest matches that were probably closely related genera Penicillium, Aureobasidium, Spongipellis, but the phylogenetic tree lacked the resolution for genus identification, whereas sequence results from the one culture were more ambiguous. The RFLP analysis could detect the presence of some cultures in mixed DNA isolates, with varying degrees of sensitivity. Although there are not yet enough published sequences to identity less common airborne fungi using sequencing of the SSU rDNA gene, our results indicate that the molecular methods evaluated in this study could have the potential to identity fungal spores from common genera and detect them in mixed environmental samples.
- Medicine and Medical Research