Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads.
Brucellosis is a highly contagious zoonosis and still a major human health problem in endemic areas of the world. Although several diagnostic tools are available, most of them are difficult to implement especially in developing countries where complex health facilities are limited. Taking advantage...
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ftdoajarticles:oai:doaj.org/article:d135551061a7487ebba667a4aedbfbdd 2023-05-15T15:16:54+02:00 Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. Andrés E Ciocchini Diego A Rey Serantes Luciano J Melli Jeremy A Iwashkiw Bettina Deodato Jorge Wallach Mario F Feldman Juan E Ugalde Diego J Comerci 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0002048 https://doaj.org/article/d135551061a7487ebba667a4aedbfbdd EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3573069?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 doi:10.1371/journal.pntd.0002048 1935-2727 1935-2735 https://doaj.org/article/d135551061a7487ebba667a4aedbfbdd PLoS Neglected Tropical Diseases, Vol 7, Iss 2, p e2048 (2013) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2013 ftdoajarticles https://doi.org/10.1371/journal.pntd.0002048 2022-12-31T11:33:11Z Brucellosis is a highly contagious zoonosis and still a major human health problem in endemic areas of the world. Although several diagnostic tools are available, most of them are difficult to implement especially in developing countries where complex health facilities are limited. Taking advantage of the identical structure and composition of the Brucella spp. and Yersinia enterocolitica O:9 O-polysaccharide, we explored the application of a recombinant Y. enterocolitica O:9-polysaccharide-protein conjugate (OAg-AcrA) as a novel antigen for diagnosis of human brucellosis. We have developed and validated an indirect immunoassay using OAg-AcrA coupled to magnetic beads. OAg-AcrA was produced and purified with high yields in Y. enterocolitica O:9 cells co-expressing the oligosaccharyltransferase PglB and the protein acceptor AcrA of Campylobacter jejuni without the need for culturing Brucella. Expression of PglB and AcrA in Y. enterocolitica resulted in the transfer of the host O-polysaccharide from its lipid carrier to AcrA. To validate the assay and determine the cutoff values, a receiver-operating characteristic analysis was performed using a panel of characterized serum samples obtained from healthy individuals and patients of different clinical groups. Our results indicate that, using this assay, it is possible to detect infection caused by the three main human brucellosis agents (B. abortus, B. melitensis and B. suis) and select different cutoff points to adjust sensitivity and specificity levels as needed. A cutoff value of 13.20% gave a sensitivity of 100% and a specificity of 98.57%, and a cutoff value of 16.15% resulted in a test sensitivity and specificity of 93.48% and 100%, respectively. The high diagnostic accuracy, low cost, reduced assay time and simplicity of this new glycoconjugate-magnetic beads assay makes it an attractive diagnostic tool for using not only in clinics and brucellosis reference laboratories but also in locations with limited laboratory infrastructure and/or minimally trained ... Article in Journal/Newspaper Arctic Human health Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 7 2 e2048 |
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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 Andrés E Ciocchini Diego A Rey Serantes Luciano J Melli Jeremy A Iwashkiw Bettina Deodato Jorge Wallach Mario F Feldman Juan E Ugalde Diego J Comerci Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Brucellosis is a highly contagious zoonosis and still a major human health problem in endemic areas of the world. Although several diagnostic tools are available, most of them are difficult to implement especially in developing countries where complex health facilities are limited. Taking advantage of the identical structure and composition of the Brucella spp. and Yersinia enterocolitica O:9 O-polysaccharide, we explored the application of a recombinant Y. enterocolitica O:9-polysaccharide-protein conjugate (OAg-AcrA) as a novel antigen for diagnosis of human brucellosis. We have developed and validated an indirect immunoassay using OAg-AcrA coupled to magnetic beads. OAg-AcrA was produced and purified with high yields in Y. enterocolitica O:9 cells co-expressing the oligosaccharyltransferase PglB and the protein acceptor AcrA of Campylobacter jejuni without the need for culturing Brucella. Expression of PglB and AcrA in Y. enterocolitica resulted in the transfer of the host O-polysaccharide from its lipid carrier to AcrA. To validate the assay and determine the cutoff values, a receiver-operating characteristic analysis was performed using a panel of characterized serum samples obtained from healthy individuals and patients of different clinical groups. Our results indicate that, using this assay, it is possible to detect infection caused by the three main human brucellosis agents (B. abortus, B. melitensis and B. suis) and select different cutoff points to adjust sensitivity and specificity levels as needed. A cutoff value of 13.20% gave a sensitivity of 100% and a specificity of 98.57%, and a cutoff value of 16.15% resulted in a test sensitivity and specificity of 93.48% and 100%, respectively. The high diagnostic accuracy, low cost, reduced assay time and simplicity of this new glycoconjugate-magnetic beads assay makes it an attractive diagnostic tool for using not only in clinics and brucellosis reference laboratories but also in locations with limited laboratory infrastructure and/or minimally trained ... |
format |
Article in Journal/Newspaper |
author |
Andrés E Ciocchini Diego A Rey Serantes Luciano J Melli Jeremy A Iwashkiw Bettina Deodato Jorge Wallach Mario F Feldman Juan E Ugalde Diego J Comerci |
author_facet |
Andrés E Ciocchini Diego A Rey Serantes Luciano J Melli Jeremy A Iwashkiw Bettina Deodato Jorge Wallach Mario F Feldman Juan E Ugalde Diego J Comerci |
author_sort |
Andrés E Ciocchini |
title |
Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
title_short |
Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
title_full |
Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
title_fullStr |
Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
title_full_unstemmed |
Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
title_sort |
development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pntd.0002048 https://doaj.org/article/d135551061a7487ebba667a4aedbfbdd |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic Human health |
genre_facet |
Arctic Human health |
op_source |
PLoS Neglected Tropical Diseases, Vol 7, Iss 2, p e2048 (2013) |
op_relation |
http://europepmc.org/articles/PMC3573069?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 doi:10.1371/journal.pntd.0002048 1935-2727 1935-2735 https://doaj.org/article/d135551061a7487ebba667a4aedbfbdd |
op_doi |
https://doi.org/10.1371/journal.pntd.0002048 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
7 |
container_issue |
2 |
container_start_page |
e2048 |
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1766347195457470464 |