Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein

Human Norwalk viruses (HuNoVs), the most common etiological agents of acute gastroenteritis, are genetically diverse RNA viruses that frequently cause mass food poisoning internationally. Although nucleic acid detection methods, such as reverse transcription–quantitative polymerase chain reaction (R...

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Autores principales: Kimura, Yayoi, Shin, Jihye, Nakai, Yusuke, Takahashi, Masaya, Ino, Yoko, Akiyama, Tomoko, Goto, Keiko, Nagata, Noriko, Yamaoka, Yutaro, Miyakawa, Kei, Kimura, Hirokazu, Ryo, Akihide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9319599/
https://www.ncbi.nlm.nih.gov/pubmed/35891395
http://dx.doi.org/10.3390/v14071416
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author Kimura, Yayoi
Shin, Jihye
Nakai, Yusuke
Takahashi, Masaya
Ino, Yoko
Akiyama, Tomoko
Goto, Keiko
Nagata, Noriko
Yamaoka, Yutaro
Miyakawa, Kei
Kimura, Hirokazu
Ryo, Akihide
author_facet Kimura, Yayoi
Shin, Jihye
Nakai, Yusuke
Takahashi, Masaya
Ino, Yoko
Akiyama, Tomoko
Goto, Keiko
Nagata, Noriko
Yamaoka, Yutaro
Miyakawa, Kei
Kimura, Hirokazu
Ryo, Akihide
author_sort Kimura, Yayoi
collection PubMed
description Human Norwalk viruses (HuNoVs), the most common etiological agents of acute gastroenteritis, are genetically diverse RNA viruses that frequently cause mass food poisoning internationally. Although nucleic acid detection methods, such as reverse transcription–quantitative polymerase chain reaction (RT-qPCR), are the gold standard for the diagnosis of norovirus infection, alternative methods are needed for the specific and sensitive viral protein detection for rapid diagnosis and surveillance. In this study, we developed a robust and high-throughput targeted proteomic assay workflow to directly detect the VP1 major capsid protein of HuNoVs. A parallel reaction monitoring (PRM) assay using a high-resolution mass spectrometer was used to detect representative peptides derived from VP1 in six different HuNoV genotypes. An optimized protocol using synthesized heavy isotope-labeled peptides as internal standards was also used to simultaneously genotype and quantify the VP1 protein in human stool specimens. This method is expected to become a new tool for studying the molecular epidemiology of HuNoV and to shed new light on targeted proteomics in clinical practice.
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spelling pubmed-93195992022-07-27 Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein Kimura, Yayoi Shin, Jihye Nakai, Yusuke Takahashi, Masaya Ino, Yoko Akiyama, Tomoko Goto, Keiko Nagata, Noriko Yamaoka, Yutaro Miyakawa, Kei Kimura, Hirokazu Ryo, Akihide Viruses Article Human Norwalk viruses (HuNoVs), the most common etiological agents of acute gastroenteritis, are genetically diverse RNA viruses that frequently cause mass food poisoning internationally. Although nucleic acid detection methods, such as reverse transcription–quantitative polymerase chain reaction (RT-qPCR), are the gold standard for the diagnosis of norovirus infection, alternative methods are needed for the specific and sensitive viral protein detection for rapid diagnosis and surveillance. In this study, we developed a robust and high-throughput targeted proteomic assay workflow to directly detect the VP1 major capsid protein of HuNoVs. A parallel reaction monitoring (PRM) assay using a high-resolution mass spectrometer was used to detect representative peptides derived from VP1 in six different HuNoV genotypes. An optimized protocol using synthesized heavy isotope-labeled peptides as internal standards was also used to simultaneously genotype and quantify the VP1 protein in human stool specimens. This method is expected to become a new tool for studying the molecular epidemiology of HuNoV and to shed new light on targeted proteomics in clinical practice. MDPI 2022-06-28 /pmc/articles/PMC9319599/ /pubmed/35891395 http://dx.doi.org/10.3390/v14071416 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kimura, Yayoi
Shin, Jihye
Nakai, Yusuke
Takahashi, Masaya
Ino, Yoko
Akiyama, Tomoko
Goto, Keiko
Nagata, Noriko
Yamaoka, Yutaro
Miyakawa, Kei
Kimura, Hirokazu
Ryo, Akihide
Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein
title Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein
title_full Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein
title_fullStr Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein
title_full_unstemmed Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein
title_short Development of Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of Human Norovirus Major Capsid Protein
title_sort development of parallel reaction monitoring mass spectrometry assay for the detection of human norovirus major capsid protein
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9319599/
https://www.ncbi.nlm.nih.gov/pubmed/35891395
http://dx.doi.org/10.3390/v14071416
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