Multiplexed High-Throughput Serological Assay for Human Enteroviruses
Immunological assays detecting antibodies against enteroviruses typically use a single enterovirus serotype as antigen. This limits the ability of such assays to detect antibodies against different enterovirus types and to detect possible type-specific variation in antibody responses. We set out to...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2020-06-01
|
Series: | Microorganisms |
Subjects: | |
Online Access: | https://www.mdpi.com/2076-2607/8/6/963 |
id |
doaj-90f10a58955e47c58cb4289765358234 |
---|---|
record_format |
Article |
spelling |
doaj-90f10a58955e47c58cb42897653582342020-11-25T03:11:36ZengMDPI AGMicroorganisms2076-26072020-06-01896396310.3390/microorganisms8060963Multiplexed High-Throughput Serological Assay for Human EnterovirusesNiila V. V. Saarinen0Jussi Lehtonen1Riitta Veijola2Johanna Lempainen3Mikael Knip4Heikki Hyöty5Olli H. Laitinen6Vesa P. Hytönen7Faculty of Medicine and Health Technology, Tampere University, 33520 Tampere, FinlandFaculty of Medicine and Health Technology, Tampere University, 33520 Tampere, FinlandDepartment of Paediatrics, University of Oulu, 90570 Oulu, FinlandDepartment of Paediatrics, University of Turku, 20520 Turku, FinlandPediatric Research Center, Children’s Hospital, University of Helsinki and Helsinki University Hospital, 00029 Helsinki, FinlandFaculty of Medicine and Health Technology, Tampere University, 33520 Tampere, FinlandFaculty of Medicine and Health Technology, Tampere University, 33520 Tampere, FinlandFaculty of Medicine and Health Technology, Tampere University, 33520 Tampere, FinlandImmunological assays detecting antibodies against enteroviruses typically use a single enterovirus serotype as antigen. This limits the ability of such assays to detect antibodies against different enterovirus types and to detect possible type-specific variation in antibody responses. We set out to develop a multiplexed assay for simultaneous detection of antibodies against multiple enterovirus and rhinovirus types encompassing all human infecting species. Seven recombinant VP1 proteins from enteroviruses EV-A to EV-D and rhinoviruses RV-A to RV-C species were produced. Using Meso Scale Diagnostics U-PLEX platform we were able to study antibody reactions against these proteins as well as non-structural enterovirus proteins in a single well with 140 human serum samples. Adults had on average 33-fold stronger antibody responses to these antigens (<i>p</i> < 10<sup>−11</sup>) compared to children, but children had less cross-reactivity between different enterovirus types. The results suggest that this new high-throughput assay offers clear benefits in the evaluation of humoral enterovirus immunity in children, giving more exact information than assays that are based on a single enterovirus type as antigen.https://www.mdpi.com/2076-2607/8/6/963enterovirusrhinovirusmultiplex immunoassay |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Niila V. V. Saarinen Jussi Lehtonen Riitta Veijola Johanna Lempainen Mikael Knip Heikki Hyöty Olli H. Laitinen Vesa P. Hytönen |
spellingShingle |
Niila V. V. Saarinen Jussi Lehtonen Riitta Veijola Johanna Lempainen Mikael Knip Heikki Hyöty Olli H. Laitinen Vesa P. Hytönen Multiplexed High-Throughput Serological Assay for Human Enteroviruses Microorganisms enterovirus rhinovirus multiplex immunoassay |
author_facet |
Niila V. V. Saarinen Jussi Lehtonen Riitta Veijola Johanna Lempainen Mikael Knip Heikki Hyöty Olli H. Laitinen Vesa P. Hytönen |
author_sort |
Niila V. V. Saarinen |
title |
Multiplexed High-Throughput Serological Assay for Human Enteroviruses |
title_short |
Multiplexed High-Throughput Serological Assay for Human Enteroviruses |
title_full |
Multiplexed High-Throughput Serological Assay for Human Enteroviruses |
title_fullStr |
Multiplexed High-Throughput Serological Assay for Human Enteroviruses |
title_full_unstemmed |
Multiplexed High-Throughput Serological Assay for Human Enteroviruses |
title_sort |
multiplexed high-throughput serological assay for human enteroviruses |
publisher |
MDPI AG |
series |
Microorganisms |
issn |
2076-2607 |
publishDate |
2020-06-01 |
description |
Immunological assays detecting antibodies against enteroviruses typically use a single enterovirus serotype as antigen. This limits the ability of such assays to detect antibodies against different enterovirus types and to detect possible type-specific variation in antibody responses. We set out to develop a multiplexed assay for simultaneous detection of antibodies against multiple enterovirus and rhinovirus types encompassing all human infecting species. Seven recombinant VP1 proteins from enteroviruses EV-A to EV-D and rhinoviruses RV-A to RV-C species were produced. Using Meso Scale Diagnostics U-PLEX platform we were able to study antibody reactions against these proteins as well as non-structural enterovirus proteins in a single well with 140 human serum samples. Adults had on average 33-fold stronger antibody responses to these antigens (<i>p</i> < 10<sup>−11</sup>) compared to children, but children had less cross-reactivity between different enterovirus types. The results suggest that this new high-throughput assay offers clear benefits in the evaluation of humoral enterovirus immunity in children, giving more exact information than assays that are based on a single enterovirus type as antigen. |
topic |
enterovirus rhinovirus multiplex immunoassay |
url |
https://www.mdpi.com/2076-2607/8/6/963 |
work_keys_str_mv |
AT niilavvsaarinen multiplexedhighthroughputserologicalassayforhumanenteroviruses AT jussilehtonen multiplexedhighthroughputserologicalassayforhumanenteroviruses AT riittaveijola multiplexedhighthroughputserologicalassayforhumanenteroviruses AT johannalempainen multiplexedhighthroughputserologicalassayforhumanenteroviruses AT mikaelknip multiplexedhighthroughputserologicalassayforhumanenteroviruses AT heikkihyoty multiplexedhighthroughputserologicalassayforhumanenteroviruses AT ollihlaitinen multiplexedhighthroughputserologicalassayforhumanenteroviruses AT vesaphytonen multiplexedhighthroughputserologicalassayforhumanenteroviruses |
_version_ |
1724653391684042752 |