Figure 1.
Calibration of the real-time RT-PCR assay for HCV, HIV-1, SARS-CoV1, and SARS-CoV2.
We diluted purified and calibrated armored RNA with pooled normal human plasma supplemented with 1 g/L sodium azide and prepared 200-μL aliquots by 10-fold serial dilution to obtain samples containing 1010 to 101 copies. From these materials, we isolated template RNA ranging from 1010 to 101 copies (from left to right) for RT-PCR assays. Water was used as a negative control. All RNA templates were assayed in a single run using a diagnostic reagent set (Intec) for each individual virus. Real-time RT-PCR was conducted on an iCycler iQ thermal cycler (Bio-Rad).