For the second run, 1 ml of the first run product was added to 24 ml reaction volume containing, 25 pmol/ml of each primer (pGp3 and pGp4), 2. Pocket warmer LAMP (pwLAMP) was performed using a loopamp DNA amplification kit (Eiken Chemical) described previously [32] . Each 25 ml reaction mixture contained 1.6 mM each of FIP and BIP, 0.2 mM each of F3 and B3, 0.8 mM each of LF and LB, 26 reaction mixture (12.5 ml), 1 ml of Bst DNA polymerase, 1 ml of fluorescence detection reagent (Eiken Chemical), 3.5 ml distilled water and 1 ml sample.