Superior sensitivity for viral RNA detection.
- Superior Sensitivity – Higher yields optimised for lower viral RNA inputs
- Tough Tested - Tolerant to a wide range of PCR inhibitors
- Multiplexing – Supports highly sensitive detection for up to four targets
- Broad Dynamic Range – Reliable data from your precious samples every time
qScript™ 1-Step Virus ToughMix is a 2x, ready-to-use, master mix for rapid detection of RNA viruses such as Flu-A, Flu-B, and SARS-CoV-2, using one-step, or single-tube reverse transcription quantitative PCR (RT-qPCR). It has been optimised for maximum sensitivity to enable reliable quantification of very low input quantities of RNA using dual-labelled hydrolysis probe detection chemistries such as TaqMan® probes in single or multiplexed assay formats. qScript 1-Step Virus ToughMix contains all required components for RT-qPCR except RNA template and primer/probe assays.
qScript™ 1-Step Virus ToughMix is powered by an engineered reverse transcriptase with reduced RNase H activity and improved activity and stability at higher temperatures, that includes a RT Hot-Start technology to suppresses non-specific primer extension by the RT before cDNA synthesis. The use of higher temperatures (50 to 55 °C) for the first-strand step of one-step RT-qPCR provides higher specificity for primer annealing and disruption of RNA secondary structure that can interfere with cDNA synthesis. These features combined with a stringent mAb hot-start Taq polymerase and ToughMix reagent technology to neutralize many common PCR inhibitors, provide reproducible low-copy quantification as well as extended room temperature stability of fully assembled reactions for automated reaction assembly. qScript™ 1-Step Virus ToughMix does not contain an internal reference dye.
qScript™ 1-step Virus ToughMix was benchmarked against the gold standard qScript™ XLT 1-Step RT-qPCR ToughMix for detection of varying amounts of SARS-CoV-2 RNA in a background of human total RNA. Multiplexed RT-qPCR of the E-gene (FAM), RdRp (HEX) and RNase P Internal Control (ROX) was carried out on the Q qPCR cycler: 10 min at 50 °C; 1 min at 95 °C; followed by 45 cycles of 5 sec at 95 °C; 10 sec at 60 °C. Both products generated excellent results. However, the qScript 1-Step Virus ToughMix demonstrated earlier detection and robust amplification curves (higher signal amplitude) for low copy E-gene detection. Quantabio recommends both of these high performing products for RNA virus detection. While highly similar in composition, the improved specificity provided by the reverse transcription hot-start of the qScript™ 1-Step Virus ToughMix often translates to improved sensitivity and fewer false negative calls at or near the limit of detection (LOD), particularly in multiplexed assays.