Bacterial gene expression detected in human faeces by reverse transcription-PCR

N.A. Fitzsimons, A.D.L. Akkermans, W.M. de Vos, E.E. Vaughan

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)

Abstract

A method to isolate and specifically detect bacterial messenger RNA (mRNA) in human faeces is presented. The surface layer protein gene slpA of Lactobacillus acidophilus ATCC 4356(T) was chosen as a model system because it is transcribed at a high level to a relatively stable mRNA (Boot et al., 1996, J. Bacteriol. 178, 5388-5394). A simulation of the recovery of bacterial cells in the faecal ecosystem was achieved by spiking faecal homogenates with different levels of L. acidophilus cells and total RNA was isolated using a method based on Macaloid clay. The slpA transcript could be detected by reverse transcription-PCR (RT-PCR) when the L. acidophilus cells comprised more than 0.14% (similar to2 x 10(7) cells g(-1) faeces) of the complex faecal community. (C) 2003 Elsevier Science B.V. All rights reserved.
Original languageEnglish
Pages (from-to)133-140
JournalJournal of Microbiological Methods
Volume55
DOIs
Publication statusPublished - 2003

Keywords

  • 16s ribosomal-rna
  • gradient gel-electrophoresis
  • in-situ hybridization
  • human fecal samples
  • layer protein gene
  • messenger-rna
  • lactobacillus-acidophilus
  • gastrointestinal-tract
  • chain-reaction
  • rt-pcr

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