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Coutard, Francois; Pommepuy, Monique; Loaec, Solen; Hervio Heath, Dominique. |
Aims: This work investigates the maintenance of viability and potential virulence of Vibrio parahaemolyticus in a viable but nonculturable population (VBNC) state by reverse transcription-polymerase chain reaction (RT-PCR). Methods and Results: Housekeeping genes, 16S-23S rDNA and rpoS, as well as virulence genes, tdh1 and tdh2, were selected and detected by PCR in a pathogenic strain of V. parahaemolyticus (Vp4). Their expression was then studied by RT-PCR in V. parahaemolyticus Vp4 cultivated in rich medium at 37degreesC. The 16S-23S rDNA and rpoS, tdh1, tdh2 genes were transcripted at the mid-logarithmic, stationary and late stationary phases, corresponding to various physiological states. The expression of these genes was also studied by RT-PCR in a... |
Tipo: Text |
Palavras-chave: Vibrio parahaemolyticus; Viable but nonculturable state; Tdh2; Tdh1; RT PCR; RpoS; Environment; 16S 23S rDNA. |
Ano: 2005 |
URL: http://archimer.ifremer.fr/doc/2005/publication-1098.pdf |
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Coutard, Francois; Lozach, Solen; Pommepuy, Monique; Hervio Heath, Dominique. |
A real-time reverse transcription-PCR method was developed to determine whether the recovery of culturability of viable but nonculturable (VBNC) Vibrio parahaemolyticus induced the expression of virulence genes coding for the thermostable direct hemolysin and for type III secretion system 2 (TTSS2). The culturability of clinical strain Vp5 of V. parahaemolyticus in artificial seawater at 4°C was monitored, and the VBNC state was obtained. One day after entry into the VBNC state, temperature upshifts to 20 and 37°C allowed the recovery of culturability. Standard curves for the relative quantification of expression of the housekeeping genes rpoS, pvsA, fur, and pvuA; the tdh2 gene; and the TTSS2 genes (VPA1354, VPA1346, and VPA1342) were established. The... |
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Ano: 2007 |
URL: http://archimer.ifremer.fr/doc/2007/publication-2787.pdf |
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