RT-qPCR Assay Optimised for Multiplexed SARS-CoV-2 and Winter Virus Tests

Multiplex assays, ideal for SARS-CoV-2 and winter virus testing, have the potential to save considerable time, effort and reagent use. However, they are complex to optimise – so PCR Biosystems has done the hard work for you in a new application note detailing a rapid and sensitive multiplex RT-qPCR assay for SARS-CoV-2 detection. This is designed to contribute to, and support efforts in, developing effective high-throughput diagnostic testing for COVID-19.

SARS-CoV-2 testing during the winter months will likely form part of a multiplexed respiratory panel test designed to distinguish SARS-CoV-2 from other common seasonal viruses such as Influenza A, Influenza B and Respiratory Syncytial Virus (RSV).

It is well recognised that multiplex approaches bring several financial and efficiency gains by allowing for several tests to be performed simultaneously on a single sample.  However, they are harder to optimise given the increased number of targets in just one RT-qPCR reaction.

To help those looking to use multiplex RT-qPCR assays for laboratory-based tests or for the development of diagnostic kits for viruses such as SARS-CoV-2, RSV or influenza, PCR Biosystems has published a new application note that sets out a proven experimental set-up for successful SARS-CoV-2 detection in a multiplex format.

The application note describes the optimal conditions for achieving a sensitive result, including the RT-qPCR variables of reverse transcription time and temperature, as well as concentrations of primers and probes. Using an optimised set-up, PCR Biosystems demonstrates the ability to detect SARS-CoV-2 at concentrations as low as four viral copies per reaction.

To aid the development of respiratory virus panels, PCR Biosystems has recently launched qPCRBIO Probe 1-Step Virus Detect. Optimised with a high-concentration 4X mix, this kit allows more sample to be added to boost analytical sensitivity, even when working with low-volume reactions. The qPCRBIO Probe 1-Step Virus Detect kit is ideally suited to high-throughput, highly multiplexed virus detection assays, such as the one outlined in the application note.

To download the application note, please click here.