The real-time PCR step was immediately followed by real-time differential melting curve analysis using the SmartCycler TM machine. DNA melting was performed immediately following PCR on the Smart Cycler I machine. Samples were heated from 708C to 958C at 0.18C/s. Differential fluorescent intensity curves (ÀdF/dT) were smoothed and used for identification of melting peak (s).