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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.
Schuster, Ofir; Atiya-Nasagi, Yafit; Rosen, Osnat; Zvi, Anat; Glinert, Itai; Ben Shmuel, Amir; Weiss, Shay; Laskar, Orly; Feldberg, Liron.
Afiliación
  • Schuster O; Department of Infectious Diseases, Israel Institute for Biological Research (IIBR), Ness Ziona, Israel. ofirsc@iibr.gov.il.
  • Atiya-Nasagi Y; Department of Infectious Diseases, Israel Institute for Biological Research (IIBR), Ness Ziona, Israel.
  • Rosen O; Department of Biotechnology, IIBR, Ness Ziona, Israel.
  • Zvi A; Department of Biochemistry and Molecular Genetics, IIBR, Ness Ziona, Israel.
  • Glinert I; Department of Infectious Diseases, Israel Institute for Biological Research (IIBR), Ness Ziona, Israel.
  • Ben Shmuel A; Department of Infectious Diseases, Israel Institute for Biological Research (IIBR), Ness Ziona, Israel.
  • Weiss S; Department of Infectious Diseases, Israel Institute for Biological Research (IIBR), Ness Ziona, Israel.
  • Laskar O; Department of Infectious Diseases, Israel Institute for Biological Research (IIBR), Ness Ziona, Israel.
  • Feldberg L; Department of Analytical Chemistry, IIBR, Ness Ziona, Israel. lironf@iibr.gov.il.
Anal Bioanal Chem ; 414(5): 1949-1962, 2022 Feb.
Article en En | MEDLINE | ID: mdl-34981149
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 × 104 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.
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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Cromatografía Liquida / Separación Inmunomagnética / Espectrometría de Masas en Tándem / Prueba de COVID-19 / SARS-CoV-2 / COVID-19 Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Anal Bioanal Chem Año: 2022 Tipo del documento: Article País de afiliación: Israel

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Cromatografía Liquida / Separación Inmunomagnética / Espectrometría de Masas en Tándem / Prueba de COVID-19 / SARS-CoV-2 / COVID-19 Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Anal Bioanal Chem Año: 2022 Tipo del documento: Article País de afiliación: Israel