Pandemic Preparedness: An Easily Implemented LC-immuno-MRM-MS Winterplex Method for Respiratory Viral Screening

Reliable large-scale testing for respiratory viruses, including Influenza viruses, coronaviruses (such as SARS-CoV-2), and respiratory syncytial virus (RSV) is essential for both endemic surveillance and pandemic response. While RT-qPCR remains the current gold standard, the COVID-19 pandemic highlighted the need for alternative, scalable diagnostic platforms. Although alternative approaches like liquid chromatography coupled with mass spectrometry (LC-MS) are ideally suited for viral multiplexing, they have yet to undergo clinical validation. Here, we present a quantitative immuno-MRM assay, called Winterplex, for the simultaneous detection of Influenza A, Influenza B, RSV, and SARS-CoV-2, each targeted via one proteotypic peptide per virus. The method was validated according to ISO standards for in vitro diagnostics and benchmarked against RT-qPCR. To strengthen future pandemic preparedness, urgent investment is needed to clinically translate and expand such multiplexed mass spectrometry-based diagnostics, enabling faster and more flexible responses to emerging viral threats.