Limiting false-positive polymerase chain reaction results: detection of DNA and mRNA to differentiate viable from dead bacteria.
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.
Kobayashi
Hideo
H
Pathology and Laboratory Medicine Institute, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA.
Oethinger
Margret
M
Tuohy
Marion J
MJ
Procop
Gary W
GW
Hall
Gerri S
GS
Bauer
Thomas W
TW
D001424:Bacterial Infections / Q000175:diagnosis* / Q000382:microbiology; D018834:Chaperonin 60 / Q000235:genetics; D053487:DEAD-box RNA Helicases; D004269:DNA, Bacterial / Q000235:genetics*; D003951:Diagnostic Errors; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D050296:Microbial Viability; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012329:RNA, Bacterial / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D018834:Chaperonin 60; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S; C088840:deaD protein, E coli; D053487:DEAD-box RNA Helicases