Figure 3
Molecular beacons can detect DNA between 1 and 106B. microti in a duplex assay in the presence of human DNA. Amplification plots of BmTPK and ACTA1 genes in PCR assays using the human DNA representing 105 ACTA1 copies spiked with ten-fold dilutions from 1 to 106 of B. microti DNA copies were used to estimate quantities of B. microti (A) and human (C) DNA by employing both BmTPK and ACTA1 molecular beacons. The assay quantified amplicons from both the BmTPK and the ACTA1 genes in the same PCR assay tubes. A high coefficient of correlation (r2 = 0.993) between the Ct values and the parasite numbers obtained from the standard curve (B) indicates that the molecular beacons can be used effectively to quantify the parasite burden in the infected human cells using multiplex assay system using the optimized conditions.