The real time qPCR was performed with the following conditions: denaturation (94°C for 15 min), amplification and quantification (35 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 30 s), and final extension at 72°C for 5 min. The melting curve program was 52°C to 95°C, 0.2°C/read, and a 1-s hold.