RESUMEN
Here we describe a protocol for identifying metabolites in respiratory specimens of patients that are SARS-CoV-2 positive, SARS-CoV-2 negative, or H1N1 positive. This protocol provides step-by-step instructions on sample collection from patients, followed by metabolite extraction. We use ultra-high-pressure liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) for data acquisition and describe the steps for data analysis. The protocol was standardized with specific customization for SARS-CoV-2-containing respiratory specimens. For complete details on the use and execution of this protocol, please refer to Maras et al. (2021).
Asunto(s)
COVID-19/diagnóstico , Cromatografía Líquida de Alta Presión/métodos , Metabolómica/métodos , COVID-19/metabolismo , Biología Computacional , Pruebas Diagnósticas de Rutina , Perfilación de la Expresión Génica , Técnicas Genéticas , Humanos , Subtipo H1N1 del Virus de la Influenza A/metabolismo , Subtipo H1N1 del Virus de la Influenza A/patogenicidad , Espectrometría de Masas/métodos , Metaboloma , Estándares de Referencia , SARS-CoV-2/metabolismo , SARS-CoV-2/patogenicidad , Manejo de Especímenes/métodosRESUMEN
In this protocol, we describe global proteome profiling for the respiratory specimen of COVID-19 patients, patients suspected with COVID-19, and H1N1 patients. In this protocol, details for identifying host, viral, or bacterial proteome (Meta-proteome) are provided. Major steps of the protocol include virus inactivation, protein quantification and digestion, desalting of peptides, high-resolution mass spectrometry (HRMS)-based analysis, and downstream bioinformatics analysis. For complete details on the use and execution of this profile, please refer to Maras et al. (2021).