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

doi:10.1016/S0167-7012(03)00121-0

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 journal › Article › Academic › peer-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 language	English
Pages (from-to)	133-140
Journal	Journal of Microbiological Methods
Volume	55
DOIs	https://doi.org/10.1016/S0167-7012(03)00121-0
Publication status	Published - 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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10.1016/S0167-7012(03)00121-0

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@article{d62b6defe94e4cde9c4de98bb3d61559,

title = "Bacterial gene expression detected in human faeces by reverse transcription-PCR",

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.",

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",

author = "N.A. Fitzsimons and A.D.L. Akkermans and {de Vos}, W.M. and E.E. Vaughan",

year = "2003",

doi = "10.1016/S0167-7012(03)00121-0",

language = "English",

volume = "55",

pages = "133--140",

journal = "Journal of Microbiological Methods",

issn = "0167-7012",

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TY - JOUR

T1 - Bacterial gene expression detected in human faeces by reverse transcription-PCR

AU - Fitzsimons, N.A.

AU - Akkermans, A.D.L.

AU - de Vos, W.M.

AU - Vaughan, E.E.

PY - 2003

Y1 - 2003

N2 - 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.

AB - 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.

KW - 16s ribosomal-rna

KW - gradient gel-electrophoresis

KW - in-situ hybridization

KW - human fecal samples

KW - layer protein gene

KW - messenger-rna

KW - lactobacillus-acidophilus

KW - gastrointestinal-tract

KW - chain-reaction

KW - rt-pcr

U2 - 10.1016/S0167-7012(03)00121-0

DO - 10.1016/S0167-7012(03)00121-0

M3 - Article

SN - 0167-7012

VL - 55

SP - 133

EP - 140

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

ER -

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

Abstract

Keywords

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