A sensitivity test was undertaken w

A sensitivity test was undertaken with bacterial cultures and purified genomic DNA. Reference strain V. parahaemolyticus ATCC 17802 was cultured overnight at 37 ℃ in APW. It was then 10-fold diluted, and the CFU number was determined by plate count. Meanwhile, 1 mL of each dilution was collected to extract DNA as described above.The concentration of chromosomal DNA (10 ng) in the original solution extracted from a pure culture of V. parahaemolyticus ATCC 17802 was measured using a NanoDrop 2000 (Thermo Scientific, Waltham, MA, USA) and then the DNA was 10-fold serially diluted (10 ng/μL to 10 fg/μL) in sterile distilled water. PCR amplification was performed with optimized parameters as described below.The reproducibility of the entire nested PCR assay was examined by performing in triplicate and with three biological replicates on different days.