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PMID: 9546166 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Detection of mRNA by reverse transcription-PCR as an indicator of viability in Escherichia coli cells.

Applied and environmental microbiology ·Vol. 64 ·No. 4 ·1998-04-00 ·Pages 1313-8

Sheridan GE, Masters CI, Shallcross JA, MacKey BM

Abstract

The relationship between the detection of mRNA and cellular viability in Escherichia coli was investigated in cells killed by heat or ethanol. Reverse transcription-PCR (RT-PCR) methods were developed for detecting mRNA from rpoH, groEL, and tufA genes. mRNA from all three genes was detected immediately after the cells had been killed by heat or ethanol but gradually disappeared with time when dead cells were held at room temperature. In heat-killed cells, some mRNA targets became undetectable after 2 to 16 h, whereas after ethanol treatment, mRNA was still detected after 16 h. In contrast, 16S rRNA was detected by RT-PCR in all samples containing dead cells and did not disappear during a subsequent incubation of 16 h at room temperature. Of the different types of nucleic acid, mRNA is the most promising candidate for an indicator of viability in bacteria, but its persistence in dead cells depends on the inactivating treatment and subsequent holding conditions.

MeSH Terms
Bacterial Proteins/genetics Base Sequence Chaperonin 60/genetics DNA Primers/genetics Escherichia coli/cytology,genetics,metabolism Ethanol/pharmacology Genes, Bacterial Heat-Shock Proteins/genetics Hot Temperature Polymerase Chain Reaction/methods RNA, Bacterial/genetics,metabolism RNA, Messenger/genetics,metabolism RNA, Ribosomal, 16S/genetics,metabolism Sigma Factor/genetics Transcription Factors
Chemicals
Bacterial Proteins Chaperonin 60 DNA Primers Heat-Shock Proteins RNA, Bacterial RNA, Messenger RNA, Ribosomal, 16S Sigma Factor Transcription Factors heat-shock sigma factor 32 Ethanol
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Sheridan G E
Institute of Food Research, Reading, United Kingdom.
Masters C I
Shallcross J A
MacKey B M
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.64.4.1313-1318.1998
Published
1998-04-00
Pages
1313-8
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC106147
Subset
IM
External Links
PubMed source DOI: 10.1128/AEM.64.4.1313-1318.1998 PMC Full Text View journal details
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