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DNA amplifications were performed in a Bio-Rad CFX-96 real-time thermocycler (Bio-Rad, USA), running a standard program (95°C for 2 min, 40 cycles at 95°C for 10 s, 60°C for 10 s and 72°C for 30 s, followed by a melting curve analysis to determine amplicon specificity using a temperature range from 65 to 95°C with increments of 0.5°C)

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