sterile distilled water. Thermal cycling conditions included 1 min at 95 °C followed by 40 amplification cycles of 10 s at 95 °C and 34 s at 60 °C. Fluorescence was detected during the second stage of each cycle. To evaluate amplification specificity, a melt curve analysis was performed at the end of each PCR run. A melt curve profile was obtained by continuously measuring fluorescence while heating the mixture to 95 °C, cooling to 60 °C (15 s), and slowly heating to 95 °C at 0.1 °C s−1.