The amplifications
were performed with an initial denaturation at 95 ◦C for
5min, followed by cycling stage with 40 cycles of 95 ◦C for 15 s,
60 ◦C for 20 s, and 68 ◦C for 30 s, then followed by melt curve stage
for 95 ◦C for 15 s, 60 ◦C for 60 s, 95 ◦C for 30 s, and 60 ◦C for 15 s