Effect of stress treatments on the detection of Listeria monocytogenes and enterotoxigenic Escherichia coli by the polymerase chain reaction.
The relationship between viability assessed by plate counts and detectability by gene probe-polymerase chain reaction (PCR) techniques was examined with cells of Escherichia coli and Listeria monocytogenes previously exposed to a range of stress treatments. In all cases the organisms were detectable by PCR after plate counts had declined to zero. Treatment with acid or hydrogen peroxide caused loss of PCR soon after viability was lost, but strong PCR signals were obtained from starved or desiccated cells long after cells became non-viable. Exposure to temperatures up to 100 degrees C had little effect on detection by PCR and even autoclaving cells at 121 degrees C for 15 min failed to abolish PCR detection completely. There is thus no simple relationship between viability and detectability by PCR. Detection of pathogens by PCR in environmental monitoring requires additional evidence of viability before risk can be properly assessed.
Masters
C I
CI
Institute of Food Research, Reading Laboratory, Earley Gate, Reading, UK.
Shallcross
J A
JA
Mackey
B M
BM
D001427:Bacterial Toxins; D001483:Base Sequence; D002455:Cell Division; D004768:Enterotoxins; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000302:isolation & purification* / Q000472:pathogenicity; D006358:Hot Temperature / Q000009:adverse effects; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008089:Listeria monocytogenes / Q000187:drug effects / Q000254:growth & development / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D014774:Virulence
D001427:Bacterial Toxins; D004768:Enterotoxins; D006861:Hydrogen Peroxide