{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T21:48:38Z","timestamp":1775166518369,"version":"3.50.1"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,6,9]],"date-time":"2022-06-09T00:00:00Z","timestamp":1654732800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004809","name":"Financiadora de Estudos e Projetos","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2013\/08135-2"],"award-info":[{"award-number":["2013\/08135-2"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2018\/07239-2"],"award-info":[{"award-number":["2018\/07239-2"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["301275\/2019-0"],"award-info":[{"award-number":["301275\/2019-0"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["301677\/2018-2"],"award-info":[{"award-number":["301677\/2018-2"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["465539\/2014-9"],"award-info":[{"award-number":["465539\/2014-9"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Med."],"abstract":"<jats:p>Human T cell lymphotropic virus (HTLV) is the caustive agent of two main conditions i. e., the HTLV-1-associated myelopathy\/tropical spastic paraparesis (HAM\/TSP) and the adult T-cell leukemia\/lymphoma (ATLL). HTLV diagnosis is based on serological and molecular approaches; however, an accurate and validated method is still needed. The objective of this study was to establish a rapid and sensitive molecular test to confirm and discriminate HTLV 1\/2 types. The test validation was performed as a multicentric study involving HTLV confirmation centers throughout Brazil. Proviral DNA was extracted from whole blood and the amplification was performed using in-house designed primer and probe sets targeting the <jats:italic>pol<\/jats:italic> genomic region. An internal control to validate the extraction and amplification was also included. The limit of detection (LoD) of the assay was four copies\/reaction for HTLV-1 and 10.9 copies\/reaction for HTLV-2. The diagnostic sensitivity of the platform was 94.6% for HTLV-1, 78.6% for HTLV-2, and the specificity was 100% for both viruses. Cross-reactions of the test with human viruses including HAV, HBV, HCV, HIV-1\/2, and parvovirus B19 were not observed. During the multicentric validation, the test was used to screen a total of 692 blood samples obtained from previously confirmed HTLV-positive individuals. From these, 91.1% tested positive being concordant with the previously obtained results. In conclusion, our duoplex-RT-PCR-HTLV1 \/2 presented adequate efficiency for HTLV-1\/2 differentiation showing high sensitivity and specificity. Therefore, it can be a suitable tool for confirmation of suspected and inconclusive HTLV cases, prenatal and pre-transplant diagnosis, in Brazil and in other countries HTLV-endemic countries.<\/jats:p>","DOI":"10.3389\/fmed.2022.881630","type":"journal-article","created":{"date-parts":[[2022,6,9]],"date-time":"2022-06-09T04:57:01Z","timestamp":1654750621000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Rapid and Sensitive Qualitative Duoplex Real-Time PCR Method for Discriminatory and Confirmatory Diagnosis of HTLV-1 and HTLV-2 Infections: Brazilian Multicentric Study"],"prefix":"10.3389","volume":"9","author":[{"given":"Mauricio Cristiano","family":"Rocha-Junior","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Evandra Strazza","family":"Rodrigues","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Svetoslav Nanev","family":"Slavov","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tatiane","family":"Assone","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ma\u00edra","family":"Pedreschi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Debora Glenda Lima","family":"de La Roque","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maisa","family":"Sousa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Viviana","family":"Olavarria","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernardo","family":"Galv\u00e3o-Castro","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benedito Antonio Lopes da","family":"Fonseca","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Augusto C\u00e9sar","family":"Penalva de Oliveira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jerusa","family":"Smid","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oswaldo Massaiti","family":"Takayanagui","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jorge","family":"Casseb","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dimas Tadeu","family":"Covas","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simone","family":"Kashima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,6,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"7415","DOI":"10.1073\/pnas.77.12.7415","article-title":"Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma","volume":"77","author":"Poiesz","year":"1980","journal-title":"Proc Natl Acad Sci USA."},{"key":"B2","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1126\/science.6981847","article-title":"A new subtype of human T-cell leukemia virus (HTLV-II) associated with a T-cell variant of hairy cell leukemia","volume":"218","author":"Kalyanaraman","year":"1982","journal-title":"Science"},{"key":"B3","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1186\/1742-4690-2-30","article-title":"Discovery of a new human T-cell lymphotropic virus (HTLV-3) in Central Africa","volume":"2","author":"Calattini","year":"2005","journal-title":"Retrovirology"},{"key":"B4","doi-asserted-by":"publisher","first-page":"7994","DOI":"10.1073\/pnas.0501734102","article-title":"Emergence of unique primate T-lymphotropic viruses among central African bushmeat hunters","volume":"102","author":"Wolfe","year":"2005","journal-title":"Proc Natl Acad Sci USA."},{"key":"B5","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1016\/S0140-6736(85)92734-5","article-title":"Antibodies to human T-lymphotropic virus type-I in patients with tropical spastic paraparesis","volume":"2","author":"Gessain","year":"1985","journal-title":"Lancet."},{"key":"B6","first-page":"1036","article-title":"Adult T-cell leukemia: concept and problems (author's transl)","volume":"20","author":"Takatsuki","year":"1979","journal-title":"Rinsho Ketsueki."},{"key":"B7","doi-asserted-by":"publisher","first-page":"S50","DOI":"10.1016\/S0165-5728(98)91243-7","article-title":"Recent studies on the epidemiology, clinical features and pathogenic mechanisms of HTLV-I associated myelopathy (HAM\/TSP) and other diseases associated to HTLV","volume":"1","author":"Osame","year":"1997","journal-title":"J Neurovirol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"1345","DOI":"10.1016\/0140-6736(90)92896-P","article-title":"Infective dermatitis of Jamaican children: a marker for HTLV-I infection","volume":"336","author":"LaGrenade","year":"1990","journal-title":"Lancet."},{"key":"B9","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1016\/S0140-6736(89)90038-X","article-title":"Chronic inflammatory arthropathy associated with HTLV-I","volume":"1","author":"Nishioka","year":"1989","journal-title":"Lancet."},{"key":"B10","doi-asserted-by":"publisher","first-page":"388","DOI":"10.3389\/fmicb.2012.00388","article-title":"Epidemiological aspects and world distribution of HTLV-1 infection","volume":"3","author":"Gessain","year":"2012","journal-title":"Front Microbiol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"926","DOI":"10.1590\/S0102-311X2005000300027","article-title":"Heterogeneous geographic distribution of human T-cell lymphotropic viruses I and II (HTLV-I\/II): serological screening prevalence rates in blood donors from large urban areas in Brazil","volume":"21","author":"Catalan-Soares","year":"2005","journal-title":"Cad Saude Publica"},{"key":"B12","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1590\/S0036-46652012000300002","article-title":"HTLV-1\/2 seroprevalence and coinfection rate in Brazilian first-time blood donors: an 11-year follow-up","volume":"54","author":"Pinto","year":"2012","journal-title":"Rev Inst Med Trop S\u00e3o Paulo"},{"key":"B13","doi-asserted-by":"publisher","first-page":"813","DOI":"10.1089\/aid.1995.11.813","article-title":"Identification of human T cell lymphotropic virus type IIa infection in the Kayapo, an indigenous population of Brazil","volume":"11","author":"Ishak","year":"1995","journal-title":"AIDS Res Hum Retroviruses."},{"key":"B14","doi-asserted-by":"publisher","first-page":"4327","DOI":"10.1128\/JVI.72.5.4327-4340.1998","article-title":"African origin of human T-lymphotropic virus type 2 (HTLV-2) supported by a potential new HTLV-2d subtype in Congolese Bambuti Efe Pygmies","volume":"72","author":"Vandamme","year":"1998","journal-title":"J Virol."},{"key":"B15","unstructured":"Portaria n\u00b0 1376, de 19 de novembro de 1993. Normas t\u00e9cnicas para coleta, processamento e transfus\u00e3o de sangue, componentes e derivadosBrasil1993"},{"key":"B16","doi-asserted-by":"publisher","first-page":"780","DOI":"10.1046\/j.1537-2995.2002.00114.x","article-title":"Strategies for diagnosis of HTLV-I and -II","volume":"42","author":"Thorstensson","year":"2002","journal-title":"Transfusion."},{"key":"B17","doi-asserted-by":"publisher","first-page":"427","DOI":"10.1086\/499273","article-title":"Human T lymphotropic virus types I and II western blot seroindeterminate status and its association with exposure to prototype HTLV-I","volume":"193","author":"Yao","year":"2006","journal-title":"J Infect Dis"},{"key":"B18","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1016\/j.jcv.2011.05.022","article-title":"Multiplex real-time PCR for the detection and quantitation of HTLV-1 and HTLV-2 proviral load: addressing the issue of indeterminate HTLV results","volume":"52","author":"Waters","year":"2011","journal-title":"J Clin Virol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/S0166-0934(01)00445-1","article-title":"Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay","volume":"102","author":"Deh\u00e9e","year":"2002","journal-title":"J Virol Methods."},{"key":"B20","doi-asserted-by":"publisher","first-page":"12433","DOI":"10.1002\/jcp.28087","article-title":"Current approaches for detection of human T-lymphotropic virus type 1: a systematic review","volume":"234","author":"Fani","year":"2019","journal-title":"J Cell Physiol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"559","DOI":"10.1128\/CMR.13.4.559","article-title":"Multiplex PCR: optimization and application in diagnostic virology","volume":"13","author":"Elnifro","year":"2000","journal-title":"Clin Microbiol Rev"},{"key":"B22","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/S0166-0934(98)00087-1","article-title":"Quantification of human T-cell lymphotropic virus type 1 proviral load by quantitative competitive polymerase chain reaction","volume":"75","author":"Albrecht","year":"1998","journal-title":"J Virol Methods"},{"key":"B23","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1089\/aid.2012.0132","article-title":"Human T cell lymphotropic virus type 1 viral load variability and long-term trends in asymptomatic carriers and in patients with human T cell lymphotropic virus type 1-related diseases","volume":"29","author":"Demontis","year":"2013","journal-title":"AIDS Res Hum Retroviruses."},{"key":"B24","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1007\/s13365-020-00924-2","article-title":"Monitoring of HTLV-1-associated diseases by proviral load quantification using multiplex real-time PCR","volume":"28","author":"Rodrigues","year":"2022","journal-title":"J Neurovirol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/S0890-8508(03)00002-1","article-title":"Multiplex PCR using real time DNA amplification for the rapid detection and quantitation of HTLV I or II","volume":"17","author":"Estes","year":"2003","journal-title":"Mol Cell Probes"},{"key":"B26","doi-asserted-by":"publisher","first-page":"1193","DOI":"10.1099\/0022-1317-77-6-1193","article-title":"Complete nucleotide sequence of the Italian human T-cell lymphotropic virus type II isolate Gu and phylogenetic identification of a possible origin of South European epidemics","volume":"77","author":"Salemi","year":"1996","journal-title":"J Gen Virol"},{"key":"B27","doi-asserted-by":"publisher","first-page":"3682","DOI":"10.1128\/JCM.00781-09","article-title":"Development and validation of a multiplex real-time PCR assay for simultaneous genotyping and human T-lymphotropic virus type 1, 2, and 3 proviral load determination","volume":"47","author":"Moens","year":"2009","journal-title":"J Clin Microbiol"},{"key":"B28","doi-asserted-by":"publisher","first-page":"1129","DOI":"10.1128\/JCM.02006-08","article-title":"One-step, multiplex, real-time PCR assay with molecular beacon probes for simultaneous detection, differentiation, and quantification of human T-cell leukemia virus types 1, 2, and 3","volume":"47","author":"Besson","year":"2009","journal-title":"J Clin Microbiol"},{"key":"B29","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1590\/S0037-86822010000200001","article-title":"Evaluation of the use of real-time PCR for human T cell lymphotropic virus 1 and 2 as a confirmatory test in screening for blood donors","volume":"43","author":"Andrade","year":"2010","journal-title":"Rev Soc Bras Med Trop"},{"key":"B30","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1016\/j.virusres.2008.01.015","article-title":"Low DNA HTLV-2 proviral load among women in S\u00e3o Paulo City","volume":"135","author":"Montanheiro","year":"2008","journal-title":"Virus Res"}],"container-title":["Frontiers in Medicine"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmed.2022.881630\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,6,9]],"date-time":"2022-06-09T04:57:08Z","timestamp":1654750628000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmed.2022.881630\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,9]]},"references-count":30,"alternative-id":["10.3389\/fmed.2022.881630"],"URL":"https:\/\/doi.org\/10.3389\/fmed.2022.881630","relation":{},"ISSN":["2296-858X"],"issn-type":[{"value":"2296-858X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,9]]},"article-number":"881630"}}