{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:02:19Z","timestamp":1760144539307,"version":"build-2065373602"},"reference-count":82,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2024,4,25]],"date-time":"2024-04-25T00:00:00Z","timestamp":1714003200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Banco Nacional de Desenvolvimento Econ\u00f4mico e Social","award":["15.2.0473.1","10.571.5020.20QF.0001"],"award-info":[{"award-number":["15.2.0473.1","10.571.5020.20QF.0001"]}]},{"name":"National Health Fund","award":["15.2.0473.1","10.571.5020.20QF.0001"],"award-info":[{"award-number":["15.2.0473.1","10.571.5020.20QF.0001"]}]},{"name":"Funda\u00e7\u00e3o de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico em Sa\u00fade (FIOTEC)","award":["15.2.0473.1","10.571.5020.20QF.0001"],"award-info":[{"award-number":["15.2.0473.1","10.571.5020.20QF.0001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Trachoma is the world-leading infectious cause of preventable blindness and is caused by the bacteria Chlamydia trachomatis. In developing countries, diagnosis is usually based on clinical evaluation. Serological-based tests are cheaper than molecular-based ones, but the latter are more sensitive and specific. The present study developed a new duplex qPCR which concomitantly detects the C. trachomatis cryptic plasmid and the human 18S rRNA gene, with an LOD95% for C. trachomatis DNA of 13.04 genome equivalents per reaction. The new qPCR was tested using 50 samples from an endemic area and 12 from a non-endemic area that were previously characterized using direct immunofluorescence assay (DFA) and clinical evaluation. Among the 50 endemic samples, 3 were found to be positive by clinical evaluation (6%), 18 were found to be positive by DFA (36%), and 48 were found to be positive by qPCR (96%). Next, the new duplex qPCR was validated using 50 samples previously characterized by qPCR. Validation was carried out on a benchtop instrument (ABI7500) or on a portable point-of-care instrument (Q3-Plus), showing 95% specificity and 100% sensitivity. The ubiquitous presence of C. trachomatis DNA in samples from the endemic region confirms that constant monitoring is of paramount importance for the effective measurement of the elimination of trachoma. The newly developed duplex qPCR presented in this study, along with its validation in a portable qPCR system, constitutes important tools toward achieving this goal.<\/jats:p>","DOI":"10.3390\/diagnostics14090892","type":"journal-article","created":{"date-parts":[[2024,4,26]],"date-time":"2024-04-26T10:56:32Z","timestamp":1714128992000},"page":"892","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Validation of a New Duplex Real-Time Polymerase Chain Reaction for Chlamydia trachomatis DNA Detection in Ocular Swab Samples"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7926-7952","authenticated-orcid":false,"given":"Joana da Felicidade Ribeiro","family":"Favacho","sequence":"first","affiliation":[{"name":"Evandro Chagas Institute, Secretariat of Health and Environment Surveillance, Ministry of Health (IEC\/SVSA\/MS), Ananindeua 67030-000, PA, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keren Kariene","family":"Leite","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology of Paran\u00e1 (IBMP), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1348-1523","authenticated-orcid":false,"given":"Thiago","family":"Jacomasso","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology of Paran\u00e1 (IBMP), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aline Burda","family":"Farias","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology of Paran\u00e1 (IBMP), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5648-4118","authenticated-orcid":false,"given":"Luciano Chaves","family":"Franco Filho","sequence":"additional","affiliation":[{"name":"Evandro Chagas Institute, Secretariat of Health and Environment Surveillance, Ministry of Health (IEC\/SVSA\/MS), Ananindeua 67030-000, PA, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7803-2860","authenticated-orcid":false,"given":"Samara Tatielle Monteiro","family":"Gomes","sequence":"additional","affiliation":[{"name":"Evandro Chagas Institute, Secretariat of Health and Environment Surveillance, Ministry of Health (IEC\/SVSA\/MS), Ananindeua 67030-000, PA, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6513-3937","authenticated-orcid":false,"given":"Herald Souza","family":"dos Reis","sequence":"additional","affiliation":[{"name":"Evandro Chagas Institute, Secretariat of Health and Environment Surveillance, Ministry of Health (IEC\/SVSA\/MS), Ananindeua 67030-000, PA, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gardene Dourado","family":"Mota","sequence":"additional","affiliation":[{"name":"Evandro Chagas Institute, Secretariat of Health and Environment Surveillance, Ministry of Health (IEC\/SVSA\/MS), Ananindeua 67030-000, PA, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pedro Henrique de Caires","family":"Schluga","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology of Paran\u00e1 (IBMP), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Walleyd Sami","family":"Tassi","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology of Paran\u00e1 (IBMP), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rita de C\u00e1ssia Pontello","family":"Rampazzo","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology of Paran\u00e1 (IBMP), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheila Kay","family":"West","sequence":"additional","affiliation":[{"name":"Dana Center for Preventative Ophthalmology, Johns Hopkins University, Baltimore, MD 21287, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charlotte Ann","family":"Gaydos","sequence":"additional","affiliation":[{"name":"International Sexually Transmitted Disease Research Laboratory, Division of Infectious Diseases, Johns Hopkins University, Baltimore, MD 21218, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio Jos\u00e9 Ledo Alves","family":"da Cunha","sequence":"additional","affiliation":[{"name":"Institute of Studies in Public Health, Federal University of Rio de Janeiro, (IESC\/UFRJ), Rio de Janeiro 21941-592, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8434-2152","authenticated-orcid":false,"given":"Alexandre Dias Tavares","family":"Costa","sequence":"additional","affiliation":[{"name":"Carlos Chagas Institute, Funda\u00e7\u00e3o Oswaldo Cruz (FIOCRUZ PR), Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,25]]},"reference":[{"key":"ref_1","unstructured":"Brazil, Ministry of Health (2016). 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