Amplifications were carried out in a Thermocycler (MiniCyclerTM, MJ Research-Watertown, MA, USA) using the following conditions:initial denaturation at 95 for 5 min followed by 30 cycles of denaturation at 95 for 15 s, annealing at 61 for 15 s, and extension at 72 for 30 s, with a final extension at 72 for 8 min.