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Direct recombinase polymerase amplification assay

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  • The results showed that the RPA reaction, without requiring complex thermal cycles, was well-performed in the optimal conditions of 39°C within only 25 minutes. The limit of detection was identified as 310 fg of L. monocytogenes genomic DNA, which was 1000-fold more sensitive than the conventional PCR. RPA also succeeded to directly detect L. monocytogenes cells at a concentration as low as 2.5 × 101 Colony Forming Unit (CFU)/mL in pure cultures. In addition, RPA could accurately detect L. monocytogenes at 2.5 × 102 CFU/mL in milk without sample extraction or processing.

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