Bacterial quantification
DENTAL PLAQUE
Quantitative polymerase chain reaction (qPCR) was used to quantify dental plaque bacteria. qPCR amplification was performed using the CFX96 system (Bio-Rad, Hercules, CA). The 25-μl reaction mixture contained the SYBR reaction mix (TaKaRa, Kusatsu, Japan), template DNA (100 ng), and forward and reverse primers (500 nM each). Thermal cycling conditions were as follows: initial denaturation at 95 °C for 30 s, followed by 40 cycles each consisting of 95 °C for 15 s, and 60 °C for 30 s. Threshold cycle (CT) values were determined, and the relative abundance was calculated based on the 2