Abstract
Loop-mediated isothermal amplification (LAMP) combined with the ethidium monoazide (EMA) treatment was applied for differentiation from VBNC state and dead state of V. parahaemolyticus. The tlh gene can be amplified remarkably within 1 h when the amount of DNA was 9fg in EMA-LAMP assay. EMA has the ability to penetrate selectively through the damaged cytoplasmic membrane of dead cells and to intercalate into DNA, so the gene of the dead cells could not be amplified. Amplification of DNA from dead bacterial was successfully inhibited by EMA, whereas the DNA derived from V. parahaemolyticus in VBNC state was readily amplified.
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