Then, 1 M of eacholigonucleotide primer and 1 L of DNA were added. The amplification profile involved an initial denaturation step at 95◦C for 15 min,followed by 30 cycles at 94◦C for 30 s, 50◦C for 40 s and 72◦C for 60 s. The positive and negative controls for the extraction and PCR were included. The PCR reactions were processed in a Mastercycler Personal (Eppendorf, Milan, Italy). All reactions were performed in duplicate.