Amplification was conducted in temperature gradient thermal cycler (Biometra-T300, Gottingen, Germany) in a volume of 25µl. Each 25µl PCR mixture consisted of 1µl of bacterial DNA, 0.5 µl (10pM) of each oligodeoxynucleotide primers, 12.5 µl of 2× Master Mix Red (Ampliqon, Denmark) and 11µl DNase and RNase free water. PCR was used for detection nuc, mecA, icaA, icaD, icaB,icaC,scn, sea, sak, sep, chb genes. All primers and programs can be found in Table I [14 (link), 18 (link), 26 (link), 28-30 (link)]. After amplification, the PCR products were analyzed by electrophoresis on 1.5% agarose gel in 0.5×TBE buffer (5.4 g Tris base, 2.75 g boric acid, 2 ml 0.5 M EDTA, in 1 L). DNA ladder was a ready to use plasmid double digest sized range 100-3000bp obtained from SMOBIO Technology (Hsinchu, Taiwan). Specificity of the primers was checked by Primer Quest software tool (http://www.ncbi.nlm.nih.gov/GenBank).