Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts.
Background The Hemagglutination assay (HA) is widely used in plague diagnosis, however, it has a subjective interpretation and demands high amounts of antigen and other immunobiological supplies. On the other hand, the conventional Anti-IgG ELISA is limited by the need of specific conjugates for mul...
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ftdoajarticles:oai:doaj.org/article:51613430df304b438a34953b75593cc7 2023-05-15T15:15:47+02:00 Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. Matheus Filgueira Bezerra Camila Cavalcanti Xavier Alzira Maria Paiva de Almeida Christian Robson de Souza Reis 2022-05-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0009805 https://doaj.org/article/51613430df304b438a34953b75593cc7 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0009805 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009805 https://doaj.org/article/51613430df304b438a34953b75593cc7 PLoS Neglected Tropical Diseases, Vol 16, Iss 5, p e0009805 (2022) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2022 ftdoajarticles https://doi.org/10.1371/journal.pntd.0009805 2022-12-30T23:16:51Z Background The Hemagglutination assay (HA) is widely used in plague diagnosis, however, it has a subjective interpretation and demands high amounts of antigen and other immunobiological supplies. On the other hand, the conventional Anti-IgG ELISA is limited by the need of specific conjugates for multiple plague hosts, which leaves a gap for new diagnostic methods able to cover both the diagnosis of human cases and the epidemiological surveillance of multiple sentinel species. Methods We developed an ELISA Protein A-peroxidase method to detect anti-F1 antibodies across several species, including humans. To determine the cut-off and performance rates, HA results from 288 samples (81 rabbits, 64 humans, 66 rodents and 77 dogs) were used as reference. Next, we evaluated the agreement between Protein A-ELISA and Anti-IgG ELISA in an expanded sample set (n = 487). Results Optimal conditions were found with 250ng/well of F1 and 1:500 serum dilution. Protein A-ELISA showed high repeatability and reproducibility. We observed good correlation rates between the Protein A and IgG ELISAs optical densities and a higher positive/negative OD ratio for the Protein A-ELISA method. The overall sensitivity, specificity and area under the curve for Protein A-ELISA were 94%, 99% and 0.99, respectively. Similar results were observed for each species separately. In the analysis of the expanded sample set, there was a strong agreement between Protein A and IgG assays (kappa = 0.97). Furthermore, there was no cross-reaction with other common infectious diseases, such as dengue, Zika, Chagas disease, tuberculosis (humans) and ehrlichiosis, anaplasmosis and leishmaniasis (dogs). Conclusions Altogether, the Protein A-ELISA showed high performance when compared both to HA and Anti-IgG ELISA, with a polyvalent single protocol that requires reduced amounts of antigen and can be employed to any plague hosts. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 16 5 e0009805 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Matheus Filgueira Bezerra Camila Cavalcanti Xavier Alzira Maria Paiva de Almeida Christian Robson de Souza Reis Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Background The Hemagglutination assay (HA) is widely used in plague diagnosis, however, it has a subjective interpretation and demands high amounts of antigen and other immunobiological supplies. On the other hand, the conventional Anti-IgG ELISA is limited by the need of specific conjugates for multiple plague hosts, which leaves a gap for new diagnostic methods able to cover both the diagnosis of human cases and the epidemiological surveillance of multiple sentinel species. Methods We developed an ELISA Protein A-peroxidase method to detect anti-F1 antibodies across several species, including humans. To determine the cut-off and performance rates, HA results from 288 samples (81 rabbits, 64 humans, 66 rodents and 77 dogs) were used as reference. Next, we evaluated the agreement between Protein A-ELISA and Anti-IgG ELISA in an expanded sample set (n = 487). Results Optimal conditions were found with 250ng/well of F1 and 1:500 serum dilution. Protein A-ELISA showed high repeatability and reproducibility. We observed good correlation rates between the Protein A and IgG ELISAs optical densities and a higher positive/negative OD ratio for the Protein A-ELISA method. The overall sensitivity, specificity and area under the curve for Protein A-ELISA were 94%, 99% and 0.99, respectively. Similar results were observed for each species separately. In the analysis of the expanded sample set, there was a strong agreement between Protein A and IgG assays (kappa = 0.97). Furthermore, there was no cross-reaction with other common infectious diseases, such as dengue, Zika, Chagas disease, tuberculosis (humans) and ehrlichiosis, anaplasmosis and leishmaniasis (dogs). Conclusions Altogether, the Protein A-ELISA showed high performance when compared both to HA and Anti-IgG ELISA, with a polyvalent single protocol that requires reduced amounts of antigen and can be employed to any plague hosts. |
format |
Article in Journal/Newspaper |
author |
Matheus Filgueira Bezerra Camila Cavalcanti Xavier Alzira Maria Paiva de Almeida Christian Robson de Souza Reis |
author_facet |
Matheus Filgueira Bezerra Camila Cavalcanti Xavier Alzira Maria Paiva de Almeida Christian Robson de Souza Reis |
author_sort |
Matheus Filgueira Bezerra |
title |
Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. |
title_short |
Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. |
title_full |
Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. |
title_fullStr |
Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. |
title_full_unstemmed |
Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts. |
title_sort |
evaluation of a multi-species protein a-elisa assay for plague serologic diagnosis in humans and other mammal hosts. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2022 |
url |
https://doi.org/10.1371/journal.pntd.0009805 https://doaj.org/article/51613430df304b438a34953b75593cc7 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 16, Iss 5, p e0009805 (2022) |
op_relation |
https://doi.org/10.1371/journal.pntd.0009805 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009805 https://doaj.org/article/51613430df304b438a34953b75593cc7 |
op_doi |
https://doi.org/10.1371/journal.pntd.0009805 |
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PLOS Neglected Tropical Diseases |
container_volume |
16 |
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5 |
container_start_page |
e0009805 |
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1766346124800557056 |