Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26.
The clinical spectrum of human disease caused by the roundworms Toxocara canis and Toxocara cati ranges from visceral and ocular larva migrans to covert toxocariasis. The parasite is not typically recovered in affected tissues, so detection of parasite-specific antibodies is usually necessary for es...
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ftdoajarticles:oai:doaj.org/article:ff0ff80eda244542b4c6026165372019 2023-05-15T15:10:35+02:00 Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. John P Anderson Lisa N Rascoe Keith Levert Holly M Chastain Matthew S Reed Hilda N Rivera Isabel McAuliffe Bin Zhan Ryan E Wiegand Peter J Hotez Patricia P Wilkins Jan Pohl Sukwan Handali 2015-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0004168 https://doaj.org/article/ff0ff80eda244542b4c6026165372019 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4618131?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004168 https://doaj.org/article/ff0ff80eda244542b4c6026165372019 PLoS Neglected Tropical Diseases, Vol 9, Iss 10, p e0004168 (2015) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2015 ftdoajarticles https://doi.org/10.1371/journal.pntd.0004168 2022-12-31T05:31:13Z The clinical spectrum of human disease caused by the roundworms Toxocara canis and Toxocara cati ranges from visceral and ocular larva migrans to covert toxocariasis. The parasite is not typically recovered in affected tissues, so detection of parasite-specific antibodies is usually necessary for establishing a diagnosis. The most reliable immunodiagnostic methods use the Toxocara excretory-secretory antigens (TES-Ag) in ELISA formats to detect Toxocara-specific antibodies. To eliminate the need for native parasite materials, we identified and purified immunodiagnostic antigens using 2D gel electrophoresis followed by electrospray ionization mass spectrometry. Three predominant immunoreactive proteins were found in the TES; all three had been previously described in the literature: Tc-CTL-1, Tc-TES-26, and Tc-MUC-3. We generated Escherichia coli expressed recombinant proteins for evaluation in Luminex based immunoassays. We were unable to produce a functional assay with the Tc-MUC-3 recombinant protein. Tc-CTL-1 and Tc-TES-26 were successfully coupled and tested using defined serum batteries. The use of both proteins together generated better results than if the proteins were used individually. The sensitivity and specificity of the assay for detecting visceral larval migrans using Tc-CTL-1 plus Tc-TES-26 was 99% and 94%, respectively; the sensitivity for detecting ocular larval migrans was 64%. The combined performance of the new assay was superior to the currently available EIA and could potentially be employed to replace current assays that rely on native TES-Ag. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Eia ENVELOPE(7.755,7.755,63.024,63.024) PLOS Neglected Tropical Diseases 9 10 e0004168 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 John P Anderson Lisa N Rascoe Keith Levert Holly M Chastain Matthew S Reed Hilda N Rivera Isabel McAuliffe Bin Zhan Ryan E Wiegand Peter J Hotez Patricia P Wilkins Jan Pohl Sukwan Handali Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
The clinical spectrum of human disease caused by the roundworms Toxocara canis and Toxocara cati ranges from visceral and ocular larva migrans to covert toxocariasis. The parasite is not typically recovered in affected tissues, so detection of parasite-specific antibodies is usually necessary for establishing a diagnosis. The most reliable immunodiagnostic methods use the Toxocara excretory-secretory antigens (TES-Ag) in ELISA formats to detect Toxocara-specific antibodies. To eliminate the need for native parasite materials, we identified and purified immunodiagnostic antigens using 2D gel electrophoresis followed by electrospray ionization mass spectrometry. Three predominant immunoreactive proteins were found in the TES; all three had been previously described in the literature: Tc-CTL-1, Tc-TES-26, and Tc-MUC-3. We generated Escherichia coli expressed recombinant proteins for evaluation in Luminex based immunoassays. We were unable to produce a functional assay with the Tc-MUC-3 recombinant protein. Tc-CTL-1 and Tc-TES-26 were successfully coupled and tested using defined serum batteries. The use of both proteins together generated better results than if the proteins were used individually. The sensitivity and specificity of the assay for detecting visceral larval migrans using Tc-CTL-1 plus Tc-TES-26 was 99% and 94%, respectively; the sensitivity for detecting ocular larval migrans was 64%. The combined performance of the new assay was superior to the currently available EIA and could potentially be employed to replace current assays that rely on native TES-Ag. |
format |
Article in Journal/Newspaper |
author |
John P Anderson Lisa N Rascoe Keith Levert Holly M Chastain Matthew S Reed Hilda N Rivera Isabel McAuliffe Bin Zhan Ryan E Wiegand Peter J Hotez Patricia P Wilkins Jan Pohl Sukwan Handali |
author_facet |
John P Anderson Lisa N Rascoe Keith Levert Holly M Chastain Matthew S Reed Hilda N Rivera Isabel McAuliffe Bin Zhan Ryan E Wiegand Peter J Hotez Patricia P Wilkins Jan Pohl Sukwan Handali |
author_sort |
John P Anderson |
title |
Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. |
title_short |
Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. |
title_full |
Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. |
title_fullStr |
Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. |
title_full_unstemmed |
Development of a Luminex Bead Based Assay for Diagnosis of Toxocariasis Using Recombinant Antigens Tc-CTL-1 and Tc-TES-26. |
title_sort |
development of a luminex bead based assay for diagnosis of toxocariasis using recombinant antigens tc-ctl-1 and tc-tes-26. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2015 |
url |
https://doi.org/10.1371/journal.pntd.0004168 https://doaj.org/article/ff0ff80eda244542b4c6026165372019 |
long_lat |
ENVELOPE(7.755,7.755,63.024,63.024) |
geographic |
Arctic Eia |
geographic_facet |
Arctic Eia |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 9, Iss 10, p e0004168 (2015) |
op_relation |
http://europepmc.org/articles/PMC4618131?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004168 https://doaj.org/article/ff0ff80eda244542b4c6026165372019 |
op_doi |
https://doi.org/10.1371/journal.pntd.0004168 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
9 |
container_issue |
10 |
container_start_page |
e0004168 |
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1766341582222524416 |