The procedure recommended by Roche was followed, and a Light Cycler Master Kit (Roche, Applied Science, Switzerland) was used to set up the reaction (2.0 μL master mix, 1.6 μL MgCl2 1.0 μL Primer F and R, 13.4 μL H2O, 1 μL sample). Absolute quantification analysis of the GDNA was carried out with a LightCycler 480 Instrument (Roche Applied Science, Switzerland). The amplification protocol was as follows: initial denaturation for 10 min at 94 °C followed by 45 cycles of 10 s at 94 °C, 5 s at the specific annealing temperature as given in Table 1, and 16 s at 72 °C.