Rapid identification of yeasts from positive blood culture bottles by pyrosequencing

Eur J Clin Microbiol Infect Dis. 2011 Jan;30(1):21-4. doi: 10.1007/s10096-010-1045-5. Epub 2010 Aug 28.

Abstract

We have developed a rapid protocol for the identification of Candida species from positive blood cultures by combining a simple method for nucleic acid extraction and preparation using microbial storage cardboards with polymerase chain reaction (PCR) and pyrosequencing of a small region of the 18 S rRNA gene. The protocol is robust and easy to implement and can be performed in 4 h. The method was tested against a collection of clinical blood cultures. Agreement of sequence identifications with standard microbiological methods was 100%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood / microbiology*
  • Candida / classification*
  • Candida / isolation & purification*
  • Candidiasis / diagnosis*
  • Candidiasis / microbiology
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / genetics
  • Fungemia / diagnosis*
  • Fungemia / microbiology
  • Humans
  • Mycology / methods*
  • Polymerase Chain Reaction / methods
  • RNA, Fungal / genetics
  • RNA, Ribosomal, 18S / genetics
  • Sequence Analysis, DNA / methods

Substances

  • DNA, Fungal
  • DNA, Ribosomal
  • RNA, Fungal
  • RNA, Ribosomal, 18S