Limiting false-positive polymerase chain reaction results: detection of DNA and mRNA to differentiate viable from dead bacteria

Hideo Kobayashi; Margret Oethinger; Marion J Tuohy; Gary W Procop; Gerri S Hall; Thomas W Bauer

doi:10.1016/j.diagmicrobio.2009.04.004

Limiting false-positive polymerase chain reaction results: detection of DNA and mRNA to differentiate viable from dead bacteria

Diagn Microbiol Infect Dis. 2009 Aug;64(4):445-7. doi: 10.1016/j.diagmicrobio.2009.04.004.

Authors

Hideo Kobayashi¹, Margret Oethinger, Marion J Tuohy, Gary W Procop, Gerri S Hall, Thomas W Bauer

Affiliation

¹ Pathology and Laboratory Medicine Institute, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA.

PMID: 19631099
DOI: 10.1016/j.diagmicrobio.2009.04.004

Abstract

We compared 2 methods for determining Escherichia coli viability in vitro. A 16S rDNA polymerase chain reaction (PCR) assay detected bacteria irrespective of viability. A groEL mRNA reverse transcriptase PCR was positive for 72 h but later became negative. Detecting mRNA holds promise but is tedious, and groEL may not be the best target.

Publication types

Comparative Study
Evaluation Study

MeSH terms

Bacterial Infections / diagnosis*
Bacterial Infections / microbiology
Chaperonin 60 / genetics
DEAD-box RNA Helicases
DNA, Bacterial / genetics*
Diagnostic Errors*
Escherichia coli / genetics
Escherichia coli Proteins / genetics
Microbial Viability*
Polymerase Chain Reaction / methods*
Polymerase Chain Reaction / standards
RNA, Bacterial / genetics*
RNA, Messenger / genetics*
RNA, Ribosomal, 16S / genetics

Substances

Chaperonin 60
DNA, Bacterial
Escherichia coli Proteins
RNA, Bacterial
RNA, Messenger
RNA, Ribosomal, 16S
deaD protein, E coli
DEAD-box RNA Helicases