An LC-Immuno-MRM-MS Winterplex Method Development Framework for Respiratory Viral Screening.

Area of Science:

• Clinical diagnostics
• Virology
• Mass spectrometry

Background:

• Reliable large-scale testing for respiratory viruses like influenza, SARS-CoV-2, and RSV is crucial for surveillance and pandemic response.
• RT-qPCR is the gold standard, but the COVID-19 pandemic revealed the need for scalable, alternative detection platforms.
• Mass spectrometry (MS)-based methods show promise for multiplex viral detection but require clinical validation.

Purpose of the Study:

• To develop and validate a novel assay for simultaneous detection of multiple respiratory viruses.
• To evaluate the performance of the new assay against the current gold standard (RT-qPCR).
• To highlight the potential of MS-based diagnostics for future pandemic preparedness.

Main Methods:

• Development of a quantitative immuno-multiple reaction monitoring (iMRM) assay named Winterplex.
• Simultaneous detection of influenza A, influenza B, RSV, and SARS-CoV-2 using two proteotypic peptides per virus.
• Validation according to ISO standards for in vitro diagnostics and benchmarking against RT-qPCR.

Main Results:

• The Winterplex assay demonstrated successful simultaneous detection of the targeted respiratory viruses.
• The assay's performance was benchmarked against RT-qPCR, indicating its viability as an alternative detection method.
• Validation according to ISO standards confirmed its suitability for in vitro diagnostics.

Conclusions:

• The Winterplex assay provides a validated, scalable platform for multiplexed respiratory virus detection.
• Investment in translating and expanding multiplexed MS-based diagnostic methodologies is essential for enhanced pandemic preparedness.
• MS-based diagnostics offer flexibility for incorporating diverse targets, enabling rapid responses to emerging viral threats.