Dataset Information

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

ABSTRACT: 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.

ORGANISM(S): Influenza A Virus Homo Sapiens Respiratory Syncytial Virus Gallus Gallus Severe Acute Respiratory Syndrome Coronavirus 2 Influenza B Virus H5n1 Subtype

SUBMITTER: Bart Van Puyvelde

PROVIDER: PXD064656 | panorama | Wed Feb 25 00:00:00 GMT 2026

REPOSITORIES: PanoramaPublic

ACCESS DATA

Similar Datasets

Project description:There is an urgent need for robust and high-throughput methods for SARS-CoV-2 detection in suspected pa-tient samples to facilitate disease management, surveillance, and control. Although nucleic acid detection methods such as RT-PCR are the gold standard, during the current pandemic the deployment of RT-PCR tests has been extremely slow, and key reagents such as PCR primers, and RNA extraction kits are at critical shortages. Rapid point-of-care viral antigen detec-tion methods have been previously employed for the diagnosis of respiratory viruses such as influenza and respiratory syn-cytial viruses. Therefore, the direct detection of SARS-CoV-2 viral antigens in patient samples could also be used for diagno-sis of active infection and alternative methodologies for specific and sensitive viral protein detection should be explored. Targeted mass spectrometry techniques have enabled the identification and quantitation of a defined subset of pro-teins/peptides at single amino acid resolution with attomole level sensitivity and high reproducibility. Herein we report a tar-geted mass spectrometry assay for the detection of SARS- CoV-2 spike protein and nucleoprotein in a relevant biologi-cal matrix. Recombinant full-length spike protein and nucleoprotein were digested and prototypic peptides were selected for parallel reaction monitoring (PRM) quantitation using a high resolution Orbitrap instrument. A spectral library, which con-tained 7 proteotypic peptides (4 from spike protein and 3 from nucleoprotein) and the top 3 to 4 transitionsMS2 spectra, was generated and evaluated. From the original spectral library, we selected 2 best performing peptides for the final PRM assay. The assay was evaluated using mock test samples containing inactivated SARS-CoV-2 virions, added to in-vitro de-rived mucus. The PRM assay provided a limit of detection (LOD) of ~200 attomoles and a limit of quantitation (LOQ) of ~ 390 attomoles. Extrapolating from the test samples, the projected titer of virus particles necessary for detection of SARS-CoV-2 spike and nucleoprotein detection was approximately 2E5 viral particles/mL, making it an attractive alternative to RT-PCR assays. Potentially mass spectrometry-based methods for viral antigen detection may deliver higher throughput and could serve as a complementary diagnostic tool to RT-PCR. Furthermore, this assay could be used to evaluate the pres-ence of SARS-CoV-2 in archived or recently collected biological fluids, in-vitro derived research materials, and wastewater samples

Dataset Information

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

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets