Abstract:

This study was aimed to develop three sero-specific real-time RT-PCR detection systems based on Taqman-MGB probes for the assay of polioviruses in foodstuffs. Primer and Taqman-MGB probe sets based on the 2A region of poliovirus genome were designed to detect three serotypes of polioviruses separately and simultaneously. Four artificially inoculated food samples, including oyster, blueberry, lettuce and water were analyzed based on the established standard curves using three reference molecules as calibrators. Sensitivity tests suggested that at least 2 copies of the reference molecule DNA could be stably detected both in the separate and simultaneous detection systems. Good linearity with the correlation coefficient above 0.999 and high reaction efficiency (1.01 － 1.04) were observed in four standard curves employing serially diluted reference molecule DNA as the calibrator. Three serotypes of polioviruses in four inoculated samples were successfully detected using the established real-time RT-PCR detection systems. The developed real-time RT-PCR detection systems based on Taqman-MGB probes are most suitable for polioviruses detection in foodstuffs.

Key words: polioviruses, rea1-time RT-PCR, Taqman-MGB probe, foodstuffs, reference molecule