{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T06:56:50Z","timestamp":1773730610351,"version":"3.50.1"},"reference-count":40,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2010,5,1]],"date-time":"2010-05-01T00:00:00Z","timestamp":1272672000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2010,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Accurate diagnosis of canine leishmaniasis (CanL) is essential toward a more efficient control of this zoonosis, but it remains problematic due to the high incidence of asymptomatic infections. In this study, we present data on the development of enzyme-linked immunosorbent assay (ELISA)-based techniques for the detection of antibodies against the recombinant protein\n            <jats:italic>Leishmania infantum<\/jats:italic>\n            cytosolic tryparedoxin peroxidase (\n            <jats:italic>Li<\/jats:italic>\n            cTXNPx) and a comparison of the results with those employing soluble\n            <jats:italic>Leishmania<\/jats:italic>\n            antigens from promastigote or amastigote forms and the homologue recombinant protein\n            <jats:italic>L. infantum<\/jats:italic>\n            mitochondrial TXNPx (\n            <jats:italic>Li<\/jats:italic>\n            mTXNPx). Moreover, we offer an evaluation of the diagnostic potential of rK39 for CanL in the Portuguese canine population and propose an improvement to the existing ELISA-based serological techniques by combining the\n            <jats:italic>Li<\/jats:italic>\n            cTXNPx and rK39 antigens as a\n            <jats:italic>Leishmania<\/jats:italic>\n            antigen mixture (LAM). The data demonstrated that ELISAs based on soluble promastigote or amastigote antigens had generally higher levels of sensitivity for detection of antibodies in symptomatic or asymptomatic dogs than for detection of those against isolated recombinant proteins. Nevertheless, the specificities were found to be similar for all target antigens used. Importantly, the LAM-ELISA methodology improved the overall sensitivity, maintaining a high overall level of specificity. In addition, it was demonstrated that the detection of anti-LAM IgG2 can increase the accuracy of the serological diagnosis. Overall, the obtained results showed that the strategy of combining two well-defined\n            <jats:italic>Leishmania<\/jats:italic>\n            antigens,\n            <jats:italic>Li<\/jats:italic>\n            cTXNPx and rK39, proved to be a sensitive and specific improvement to current serological diagnosis of CanL, being a useful tool for the detection of both clinical and subclinical forms of canine\n            <jats:italic>Leishmania<\/jats:italic>\n            infection.\n          <\/jats:p>","DOI":"10.1128\/jcm.02402-09","type":"journal-article","created":{"date-parts":[[2010,2,18]],"date-time":"2010-02-18T02:34:25Z","timestamp":1266460465000},"page":"1866-1874","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":32,"title":["Application of an Improved Enzyme-Linked Immunosorbent Assay Method for Serological Diagnosis of Canine Leishmaniasis"],"prefix":"10.1128","volume":"48","author":[{"given":"Nuno","family":"Santare\u0301m","sequence":"first","affiliation":[{"name":"Parasite Disease Group, IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal"}]},{"given":"Ricardo","family":"Silvestre","sequence":"additional","affiliation":[{"name":"Parasite Disease Group, IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal"}]},{"given":"Lui\u0301s","family":"Cardoso","sequence":"additional","affiliation":[{"name":"Parasite Disease Group, IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal"},{"name":"Departamento de Cie\u0302ncias Veterina\u0301rias, Universidade de Tra\u0301s-os-Montes e Alto Douro, Vila Real, Portugal"}]},{"given":"Henk","family":"Schallig","sequence":"additional","affiliation":[{"name":"KIT (Koninklijk Instituut voor de Tropen\/Royal Tropical Institute), KIT Biomedical Research, 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