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

J Microbiol Methods. 2003 Oct;55(1):133-40. doi: 10.1016/s0167-7012(03)00121-0.

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% (approximately 2 x 10(7) cells g(-1) faeces) of the complex faecal community.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics*
  • Colony Count, Microbial
  • DNA, Bacterial / isolation & purification
  • Feces / microbiology*
  • Humans
  • Lactobacillus acidophilus / genetics*
  • RNA, Bacterial / analysis*
  • RNA, Messenger / analysis*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • RNA, Bacterial
  • RNA, Messenger
  • surface layer protein A, Bacteria