Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection
Abstract This paper outlines methods used to identify novel antigens for use in the development of serological assays. Specifically, we applied these methods to a neurogenic parasitic nematode of cervids called Parelaphostrongylus tenuis. This parasite is of particular concern in both wild and domes...
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ftdoajarticles:oai:doaj.org/article:a2d7e88d772649db8aa1d34a6a7d440a 2023-07-30T03:55:46+02:00 Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection Jessie Richards Stephen Kania Abigail Wilson Emily Kent Richard Gerhold 2023-07-01T00:00:00Z https://doi.org/10.1038/s41598-023-37481-7 https://doaj.org/article/a2d7e88d772649db8aa1d34a6a7d440a EN eng Nature Portfolio https://doi.org/10.1038/s41598-023-37481-7 https://doaj.org/toc/2045-2322 doi:10.1038/s41598-023-37481-7 2045-2322 https://doaj.org/article/a2d7e88d772649db8aa1d34a6a7d440a Scientific Reports, Vol 13, Iss 1, Pp 1-8 (2023) Medicine R Science Q article 2023 ftdoajarticles https://doi.org/10.1038/s41598-023-37481-7 2023-07-16T00:38:39Z Abstract This paper outlines methods used to identify novel antigens for use in the development of serological assays. Specifically, we applied these methods to a neurogenic parasitic nematode of cervids called Parelaphostrongylus tenuis. This parasite is of particular concern in both wild and domestic ungulates as it causes significant neurological signs and definitive diagnosis is only possible post-mortem, necessitating the development of serologic assays for antemortem diagnosis. Proteins extracted from P. tenuis organisms were affinity isolated using antibodies enriched from seropositive moose (Alces alces). The proteins were analyzed using mass spectrometry and liquid chromatography to obtain amino acid sequences that were then cross-referenced to open reading frames predicted from an assembled transcriptome. An antigen of interest was assessed for immunogenic epitopes and subsequently synthesized into 10-mer synthetic overlapping peptides representing these regions. These synthetic peptides were then assessed for reactivity against positive and negative moose sera and demonstrated potential use as a serological assay in diagnostic laboratories. Known negative moose sera revealed significantly lower optical density when compared to the positive samples (p < 0.05). This method serves as a pipeline for the construction of diagnostic assays of pathogens in both human and veterinary medicine. Article in Journal/Newspaper Alces alces Directory of Open Access Journals: DOAJ Articles Scientific Reports 13 1 |
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English |
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Medicine R Science Q |
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Medicine R Science Q Jessie Richards Stephen Kania Abigail Wilson Emily Kent Richard Gerhold Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection |
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Medicine R Science Q |
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Abstract This paper outlines methods used to identify novel antigens for use in the development of serological assays. Specifically, we applied these methods to a neurogenic parasitic nematode of cervids called Parelaphostrongylus tenuis. This parasite is of particular concern in both wild and domestic ungulates as it causes significant neurological signs and definitive diagnosis is only possible post-mortem, necessitating the development of serologic assays for antemortem diagnosis. Proteins extracted from P. tenuis organisms were affinity isolated using antibodies enriched from seropositive moose (Alces alces). The proteins were analyzed using mass spectrometry and liquid chromatography to obtain amino acid sequences that were then cross-referenced to open reading frames predicted from an assembled transcriptome. An antigen of interest was assessed for immunogenic epitopes and subsequently synthesized into 10-mer synthetic overlapping peptides representing these regions. These synthetic peptides were then assessed for reactivity against positive and negative moose sera and demonstrated potential use as a serological assay in diagnostic laboratories. Known negative moose sera revealed significantly lower optical density when compared to the positive samples (p < 0.05). This method serves as a pipeline for the construction of diagnostic assays of pathogens in both human and veterinary medicine. |
format |
Article in Journal/Newspaper |
author |
Jessie Richards Stephen Kania Abigail Wilson Emily Kent Richard Gerhold |
author_facet |
Jessie Richards Stephen Kania Abigail Wilson Emily Kent Richard Gerhold |
author_sort |
Jessie Richards |
title |
Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection |
title_short |
Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection |
title_full |
Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection |
title_fullStr |
Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection |
title_full_unstemmed |
Novel methods of immunogenic antigen selection for serological diagnosis of Parelaphostrongylus tenuis infection |
title_sort |
novel methods of immunogenic antigen selection for serological diagnosis of parelaphostrongylus tenuis infection |
publisher |
Nature Portfolio |
publishDate |
2023 |
url |
https://doi.org/10.1038/s41598-023-37481-7 https://doaj.org/article/a2d7e88d772649db8aa1d34a6a7d440a |
genre |
Alces alces |
genre_facet |
Alces alces |
op_source |
Scientific Reports, Vol 13, Iss 1, Pp 1-8 (2023) |
op_relation |
https://doi.org/10.1038/s41598-023-37481-7 https://doaj.org/toc/2045-2322 doi:10.1038/s41598-023-37481-7 2045-2322 https://doaj.org/article/a2d7e88d772649db8aa1d34a6a7d440a |
op_doi |
https://doi.org/10.1038/s41598-023-37481-7 |
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Scientific Reports |
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13 |
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1772809543054524416 |