Using Sequence Analysis to Identify Cultures Derived from Airborne Spores.

Abstract

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.

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Document Details

Document Type
Technical Report
Publication Date
Jan 01, 1998
Accession Number
ADA339362

Entities

People

  • Charles H. Wick
  • Erica Cline
  • Homer R. Yeh
  • Robert L. Edmonds

Organizations

  • Edgewood Chemical Biological Center

Tags

DTIC Thesaurus Topics

  • Cells
  • Fungi
  • Polymerase Chain Reaction
  • Sequence Analysis

Fields of Study

  • Biology

Readers

  • Microbial Pathology
  • Molecular Genetics