Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1

Norovirus-associated diseases are the most common foodborne illnesses worldwide. Polymerase chain reaction-based methods are the primary diagnostics for clinical samples; however, the high mutation rate of norovirus makes viral amplification and genotyping challenging. Technological advances in mass...

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Autores principales:	Pei-Yu Chu, Hui-Wen Huang, Michittra Boonchan, Yu-Chang Tyan, Kevin Leroy Louis, Kun-Mu Lee, Kazushi Motomura, Liang-Yin Ke
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Norovirus virus-like particle (VLP) M-class ultra-performance liquid chromatography system data independent mass spectrometry (MS<sup>E</sup>) VP1 protein P2-P1b region Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/8dfc6420a92b4e24989c7a6b0d16b9f9
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spelling	oai:doaj.org-article:8dfc6420a92b4e24989c7a6b0d16b9f92021-11-25T19:14:40ZMass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP110.3390/v131123321999-4915https://doaj.org/article/8dfc6420a92b4e24989c7a6b0d16b9f92021-11-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2332https://doaj.org/toc/1999-4915Norovirus-associated diseases are the most common foodborne illnesses worldwide. Polymerase chain reaction-based methods are the primary diagnostics for clinical samples; however, the high mutation rate of norovirus makes viral amplification and genotyping challenging. Technological advances in mass spectrometry (MS) make it a promising tool for identifying disease markers. Besides, the superior sensitivity of MS and proteomic approaches may enable the detection of all variants. Thus, this study aimed to establish an MS-based system for identifying and typing norovirus. We constructed three plasmids containing the major capsid protein VP1 of the norovirus GII.4 2006b, 2006a, and 2009a strains to produce virus-like particles for use as standards. Digested peptide signals were collected using a nano-flow ultra-performance liquid chromatography mass spectrometry (nano-UPLC/MS<sup>E</sup>) system, and analyzed by ProteinLynx Global SERVER and TREE-PUZZLE software. Results revealed that the LC/MS<sup>E</sup> system had an excellent coverage rate: the system detected more than 94% of amino acids of 3.61 femtomole norovirus VP1 structural protein. In the likelihood-mapping analysis, the proportions of unresolved quartets were 2.9% and 4.9% in the VP1 and S domains, respectively, which is superior to the 15.1% unresolved quartets in current PCR-based methodology. In summary, the use of LC/MS<sup>E</sup> may efficiently monitor genotypes, and sensitively detect structural and functional mutations of noroviruses.Pei-Yu ChuHui-Wen HuangMichittra BoonchanYu-Chang TyanKevin Leroy LouisKun-Mu LeeKazushi MotomuraLiang-Yin KeMDPI AGarticleNorovirusvirus-like particle (VLP)M-class ultra-performance liquid chromatography systemdata independent mass spectrometry (MS<sup>E</sup>)VP1 proteinP2-P1b regionMicrobiologyQR1-502ENViruses, Vol 13, Iss 2332, p 2332 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Norovirus virus-like particle (VLP) M-class ultra-performance liquid chromatography system data independent mass spectrometry (MS<sup>E</sup>) VP1 protein P2-P1b region Microbiology QR1-502
spellingShingle	Norovirus virus-like particle (VLP) M-class ultra-performance liquid chromatography system data independent mass spectrometry (MS<sup>E</sup>) VP1 protein P2-P1b region Microbiology QR1-502 Pei-Yu Chu Hui-Wen Huang Michittra Boonchan Yu-Chang Tyan Kevin Leroy Louis Kun-Mu Lee Kazushi Motomura Liang-Yin Ke Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1
description	Norovirus-associated diseases are the most common foodborne illnesses worldwide. Polymerase chain reaction-based methods are the primary diagnostics for clinical samples; however, the high mutation rate of norovirus makes viral amplification and genotyping challenging. Technological advances in mass spectrometry (MS) make it a promising tool for identifying disease markers. Besides, the superior sensitivity of MS and proteomic approaches may enable the detection of all variants. Thus, this study aimed to establish an MS-based system for identifying and typing norovirus. We constructed three plasmids containing the major capsid protein VP1 of the norovirus GII.4 2006b, 2006a, and 2009a strains to produce virus-like particles for use as standards. Digested peptide signals were collected using a nano-flow ultra-performance liquid chromatography mass spectrometry (nano-UPLC/MS<sup>E</sup>) system, and analyzed by ProteinLynx Global SERVER and TREE-PUZZLE software. Results revealed that the LC/MS<sup>E</sup> system had an excellent coverage rate: the system detected more than 94% of amino acids of 3.61 femtomole norovirus VP1 structural protein. In the likelihood-mapping analysis, the proportions of unresolved quartets were 2.9% and 4.9% in the VP1 and S domains, respectively, which is superior to the 15.1% unresolved quartets in current PCR-based methodology. In summary, the use of LC/MS<sup>E</sup> may efficiently monitor genotypes, and sensitively detect structural and functional mutations of noroviruses.
format	article
author	Pei-Yu Chu Hui-Wen Huang Michittra Boonchan Yu-Chang Tyan Kevin Leroy Louis Kun-Mu Lee Kazushi Motomura Liang-Yin Ke
author_facet	Pei-Yu Chu Hui-Wen Huang Michittra Boonchan Yu-Chang Tyan Kevin Leroy Louis Kun-Mu Lee Kazushi Motomura Liang-Yin Ke
author_sort	Pei-Yu Chu
title	Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1
title_short	Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1
title_full	Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1
title_fullStr	Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1
title_full_unstemmed	Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1
title_sort	mass spectrometry-based system for identifying and typing norovirus major capsid protein vp1
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/8dfc6420a92b4e24989c7a6b0d16b9f9
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Mass Spectrometry-Based System for Identifying and Typing Norovirus Major Capsid Protein VP1

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