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Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples
Recently, numerous diagnostic approaches from different disciplines have been developed for SARS-CoV-2 diagnosis to monitor and control the COVID-19 pandemic. These include MS-based assays, which provide analytical information on viral proteins. However, their sensitivity is limited, estimated to be...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8723902/ https://www.ncbi.nlm.nih.gov/pubmed/34981149 http://dx.doi.org/10.1007/s00216-021-03831-5 |
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author | Schuster, Ofir Atiya-Nasagi, Yafit Rosen, Osnat Zvi, Anat Glinert, Itai Ben Shmuel, Amir Weiss, Shay Laskar, Orly Feldberg, Liron |
author_facet | Schuster, Ofir Atiya-Nasagi, Yafit Rosen, Osnat Zvi, Anat Glinert, Itai Ben Shmuel, Amir Weiss, Shay Laskar, Orly Feldberg, Liron |
author_sort | Schuster, Ofir |
collection | PubMed |
description | Recently, numerous diagnostic approaches from different disciplines have been developed for SARS-CoV-2 diagnosis to monitor and control the COVID-19 pandemic. These include MS-based assays, which provide analytical information on viral proteins. However, their sensitivity is limited, estimated to be 5 × 10(4) PFU/ml in clinical samples. Here, we present a reliable, specific, and rapid method for the identification of SARS-CoV-2 from nasopharyngeal (NP) specimens, which combines virus capture followed by LC–MS/MS(MRM) analysis of unique peptide markers. The capture of SARS-CoV-2 from the challenging matrix, prior to its tryptic digestion, was accomplished by magnetic beads coated with polyclonal IgG-α-SARS-CoV-2 antibodies, enabling sample concentration while significantly reducing background noise interrupting with LC–MS analysis. A sensitive and specific LC–MS/MS(MRM) analysis method was developed for the identification of selected tryptic peptide markers. The combined assay, which resulted in S/N ratio enhancement, achieved an improved sensitivity of more than 10-fold compared with previously described MS methods. The assay was validated in 29 naive NP specimens, 19 samples were spiked with SARS-CoV-2 and 10 were used as negative controls. Finally, the assay was successfully applied to clinical NP samples (n = 26) pre-determined as either positive or negative by RT-qPCR. This work describes for the first time a combined approach for immuno-magnetic viral isolation coupled with MS analysis. This method is highly reliable, specific, and sensitive; thus, it may potentially serve as a complementary assay to RT-qPCR, the gold standard test. This methodology can be applied to other viruses as well. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-021-03831-5. |
format | Online Article Text |
id | pubmed-8723902 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-87239022022-01-04 Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples Schuster, Ofir Atiya-Nasagi, Yafit Rosen, Osnat Zvi, Anat Glinert, Itai Ben Shmuel, Amir Weiss, Shay Laskar, Orly Feldberg, Liron Anal Bioanal Chem Research Paper Recently, numerous diagnostic approaches from different disciplines have been developed for SARS-CoV-2 diagnosis to monitor and control the COVID-19 pandemic. These include MS-based assays, which provide analytical information on viral proteins. However, their sensitivity is limited, estimated to be 5 × 10(4) PFU/ml in clinical samples. Here, we present a reliable, specific, and rapid method for the identification of SARS-CoV-2 from nasopharyngeal (NP) specimens, which combines virus capture followed by LC–MS/MS(MRM) analysis of unique peptide markers. The capture of SARS-CoV-2 from the challenging matrix, prior to its tryptic digestion, was accomplished by magnetic beads coated with polyclonal IgG-α-SARS-CoV-2 antibodies, enabling sample concentration while significantly reducing background noise interrupting with LC–MS analysis. A sensitive and specific LC–MS/MS(MRM) analysis method was developed for the identification of selected tryptic peptide markers. The combined assay, which resulted in S/N ratio enhancement, achieved an improved sensitivity of more than 10-fold compared with previously described MS methods. The assay was validated in 29 naive NP specimens, 19 samples were spiked with SARS-CoV-2 and 10 were used as negative controls. Finally, the assay was successfully applied to clinical NP samples (n = 26) pre-determined as either positive or negative by RT-qPCR. This work describes for the first time a combined approach for immuno-magnetic viral isolation coupled with MS analysis. This method is highly reliable, specific, and sensitive; thus, it may potentially serve as a complementary assay to RT-qPCR, the gold standard test. This methodology can be applied to other viruses as well. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-021-03831-5. Springer Berlin Heidelberg 2022-01-04 2022 /pmc/articles/PMC8723902/ /pubmed/34981149 http://dx.doi.org/10.1007/s00216-021-03831-5 Text en © Springer-Verlag GmbH Germany, part of Springer Nature 2021 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Research Paper Schuster, Ofir Atiya-Nasagi, Yafit Rosen, Osnat Zvi, Anat Glinert, Itai Ben Shmuel, Amir Weiss, Shay Laskar, Orly Feldberg, Liron Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples |
title | Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples |
title_full | Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples |
title_fullStr | Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples |
title_full_unstemmed | Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples |
title_short | Coupling immuno-magnetic capture with LC–MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples |
title_sort | coupling immuno-magnetic capture with lc–ms/ms(mrm) as a sensitive, reliable, and specific assay for sars-cov-2 identification from clinical samples |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8723902/ https://www.ncbi.nlm.nih.gov/pubmed/34981149 http://dx.doi.org/10.1007/s00216-021-03831-5 |
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