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 2×, 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 neutralise many common PCR inhibitors, provides 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 minutes at 50 °C; 1 minute at 95 °C; followed by 45 cycles of 5 seconds at 95 °C; 10 seconds 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.