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Simultaneous monitoring of eight human respiratory viruses including SARS-CoV-2 using liquid chromatography-tandem mass spectrometry.
Hodgkins, Christopher; Buckton, Laura K; Walker, Gregory J; Crossett, Ben; Cordwell, Stuart J; Horvath, Andrea R; Rawlinson, William D.
  • Hodgkins C; NSW Health Pathology, Prince of Wales Hospital, Campus Centre Building, 2031, Randwick, NSW, Australia.
  • Buckton LK; School of Life and Environmental Sciences and Charles Perkins Centre, The University of Sydney, 2006, Sydney, Australia.
  • Walker GJ; NSW Health Pathology, Prince of Wales Hospital, Campus Centre Building, 2031, Randwick, NSW, Australia.
  • Crossett B; Virology Research Laboratory, SAViD, Prince of Wales Hospital, 2031, Randwick, NSW, Australia.
  • Cordwell SJ; Schools of Medical Sciences, Women's and Children's Health, Faculty of Medicine and BABS Faculty of Science, University of New South Wales, 2052, Sydney, NSW, Australia.
  • Horvath AR; Sydney Mass Spectrometry, The University of Sydney, 2006, Sydney, NSW, Australia.
  • Rawlinson WD; School of Life and Environmental Sciences and Charles Perkins Centre, The University of Sydney, 2006, Sydney, Australia.
Sci Rep ; 12(1): 13392, 2022 08 04.
Article in English | MEDLINE | ID: covidwho-1972655
ABSTRACT
Diagnosis of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection has primarily been achieved using reverse transcriptase polymerase chain reaction (RT-PCR) for acute infection, and serology for prior infection. Assay with RT-PCR provides data on presence or absence of viral RNA, with no information on virus replication competence, infectivity, or virus characterisation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is typically not used in clinical virology, despite its potential to provide supplemental data about the presence of viral proteins and thus the potential for replication-competent, transmissible virus. Using the SARS-CoV-2 as a model virus, we developed a fast 'bottom-up' proteomics workflow for discovery of target virus peptides using 'serum-free' culture conditions, providing high coverage of viral proteins without the need for protein or peptide fractionation techniques. This workflow was then applied to Coronaviruses OC43 and 229E, Influenza A/H1N1 and H3N2, Influenza B, and Respiratory Syncytial Viruses A and B. Finally, we created an LC-MS/MS method for targeted detection of the eight-virus panel in clinical specimens, successfully detecting peptides from the SARS-CoV-2 ORF9B and nucleoprotein in RT-PCR positive samples. The method provides specific detection of respiratory viruses from clinical samples containing moderate viral loads and is an important further step to the use of LC-MS/MS in diagnosis of viral infection.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Influenza, Human / Influenza A Virus, H1N1 Subtype / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Sci Rep Year: 2022 Document Type: Article Affiliation country: S41598-022-16250-y

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Influenza, Human / Influenza A Virus, H1N1 Subtype / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Sci Rep Year: 2022 Document Type: Article Affiliation country: S41598-022-16250-y