Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification

Autores
Caputo, Mariela; Trinks, Julieta; Azcurra, Marcela; Corach, Daniel
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
HTLV-1 and HTLV-2 are present in different high-risk populations, such as sexual workers and injecting drug users (IDUs). HTLV-1 is endemic in areas of Middle East, Southern Japan and Latin America, whereas HTLV-2 infection is endemic among some Native Americans and some Central African tribes. The pathogenic consequences and clinical manifestations of these two viruses differ significantly, demanding an adequate identification; therefore, proper diagnosis of HTLV-1 and 2 infection is crucial. To get a final diagnosis of HTLV-1 or 2 infection, it is recommended that positive serologic samples should be confirmed by PCR assays or western blot (WB) analysis. Thus, the aim of this study was to develop and implement a simple reaction for the rapid identification of HTLV-1 and 2. Nested real-time PCR technique followed by high resolution melting was performed based on the tax/rex sequences of HTLV-1 (M2) and HTLV-2 (MoT) cell lines perfectly discriminating between HTLV-1 from HTLV-2, by distinct melting curve profiles. The sensitivity assay of this method revealed that at least 1 viral copy of HTLV-1 or 1·5 viral copy of HTLV-2 could be amplified. Later, this method was validated using 200 blood samples from corpses. In agreement with previous epidemiological, the HTLV-1 and 2 prevalence was 1·5% (CI 95%: 0·31–4·3) and 0·5% (CI 95%: 0·013–2·75), respectively. The strategy proposed herein has some advantages over other PCR-based tests because it not only reduces considerably time and the costs of the total diagnosis but also allows detection and discrimination of HTLV-1 and 2 in the same reaction.
Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Trinks, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Medicina Traslacional e Ingeniería Biomédica - Hospital Italiano. Instituto de Medicina Traslacional e Ingeniería Biomédica.- Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional e Ingeniería Biomédica; Argentina
Fil: Azcurra, Marcela. No especifíca;
Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Materia
HIGH RESOLUTION MELTING
HTLV-1
HTLV-2
NESTED PCR
REAL-TIME PCR
RETROVIRUS
SCREENING
VIRAL DIAGNOSIS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/188064
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/188064
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identificationCaputo, MarielaTrinks, JulietaAzcurra, MarcelaCorach, DanielHIGH RESOLUTION MELTINGHTLV-1HTLV-2NESTED PCRREAL-TIME PCRRETROVIRUSSCREENINGVIRAL DIAGNOSIShttps://purl.org/becyt/ford/3.5https://purl.org/becyt/ford/3HTLV-1 and HTLV-2 are present in different high-risk populations, such as sexual workers and injecting drug users (IDUs). HTLV-1 is endemic in areas of Middle East, Southern Japan and Latin America, whereas HTLV-2 infection is endemic among some Native Americans and some Central African tribes. The pathogenic consequences and clinical manifestations of these two viruses differ significantly, demanding an adequate identification; therefore, proper diagnosis of HTLV-1 and 2 infection is crucial. To get a final diagnosis of HTLV-1 or 2 infection, it is recommended that positive serologic samples should be confirmed by PCR assays or western blot (WB) analysis. Thus, the aim of this study was to develop and implement a simple reaction for the rapid identification of HTLV-1 and 2. Nested real-time PCR technique followed by high resolution melting was performed based on the tax/rex sequences of HTLV-1 (M2) and HTLV-2 (MoT) cell lines perfectly discriminating between HTLV-1 from HTLV-2, by distinct melting curve profiles. The sensitivity assay of this method revealed that at least 1 viral copy of HTLV-1 or 1·5 viral copy of HTLV-2 could be amplified. Later, this method was validated using 200 blood samples from corpses. In agreement with previous epidemiological, the HTLV-1 and 2 prevalence was 1·5% (CI 95%: 0·31–4·3) and 0·5% (CI 95%: 0·013–2·75), respectively. The strategy proposed herein has some advantages over other PCR-based tests because it not only reduces considerably time and the costs of the total diagnosis but also allows detection and discrimination of HTLV-1 and 2 in the same reaction.Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Trinks, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Medicina Traslacional e Ingeniería Biomédica - Hospital Italiano. Instituto de Medicina Traslacional e Ingeniería Biomédica.- Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional e Ingeniería Biomédica; ArgentinaFil: Azcurra, Marcela. No especifíca;Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaWiley Blackwell Publishing, Inc2022-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/188064Caputo, Mariela; Trinks, Julieta; Azcurra, Marcela; Corach, Daniel; Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification; Wiley Blackwell Publishing, Inc; Letters in Applied Microbiology; 75; 4; 10-2022; 804-8120266-8254CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1111/lam.13752info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:00:37Zoai:ri.conicet.gov.ar:11336/188064instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:00:38.3CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
title Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
spellingShingle Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
Caputo, Mariela
HIGH RESOLUTION MELTING
HTLV-1
HTLV-2
NESTED PCR
REAL-TIME PCR
RETROVIRUS
SCREENING
VIRAL DIAGNOSIS
title_short Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
title_full Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
title_fullStr Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
title_full_unstemmed Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
title_sort Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification
dc.creator.none.fl_str_mv Caputo, Mariela
Trinks, Julieta
Azcurra, Marcela
Corach, Daniel
author Caputo, Mariela
author_facet Caputo, Mariela
Trinks, Julieta
Azcurra, Marcela
Corach, Daniel
author_role author
author2 Trinks, Julieta
Azcurra, Marcela
Corach, Daniel
author2_role author
author
author
dc.subject.none.fl_str_mv HIGH RESOLUTION MELTING
HTLV-1
HTLV-2
NESTED PCR
REAL-TIME PCR
RETROVIRUS
SCREENING
VIRAL DIAGNOSIS
topic HIGH RESOLUTION MELTING
HTLV-1
HTLV-2
NESTED PCR
REAL-TIME PCR
RETROVIRUS
SCREENING
VIRAL DIAGNOSIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.5
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv HTLV-1 and HTLV-2 are present in different high-risk populations, such as sexual workers and injecting drug users (IDUs). HTLV-1 is endemic in areas of Middle East, Southern Japan and Latin America, whereas HTLV-2 infection is endemic among some Native Americans and some Central African tribes. The pathogenic consequences and clinical manifestations of these two viruses differ significantly, demanding an adequate identification; therefore, proper diagnosis of HTLV-1 and 2 infection is crucial. To get a final diagnosis of HTLV-1 or 2 infection, it is recommended that positive serologic samples should be confirmed by PCR assays or western blot (WB) analysis. Thus, the aim of this study was to develop and implement a simple reaction for the rapid identification of HTLV-1 and 2. Nested real-time PCR technique followed by high resolution melting was performed based on the tax/rex sequences of HTLV-1 (M2) and HTLV-2 (MoT) cell lines perfectly discriminating between HTLV-1 from HTLV-2, by distinct melting curve profiles. The sensitivity assay of this method revealed that at least 1 viral copy of HTLV-1 or 1·5 viral copy of HTLV-2 could be amplified. Later, this method was validated using 200 blood samples from corpses. In agreement with previous epidemiological, the HTLV-1 and 2 prevalence was 1·5% (CI 95%: 0·31–4·3) and 0·5% (CI 95%: 0·013–2·75), respectively. The strategy proposed herein has some advantages over other PCR-based tests because it not only reduces considerably time and the costs of the total diagnosis but also allows detection and discrimination of HTLV-1 and 2 in the same reaction.
Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Trinks, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Medicina Traslacional e Ingeniería Biomédica - Hospital Italiano. Instituto de Medicina Traslacional e Ingeniería Biomédica.- Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional e Ingeniería Biomédica; Argentina
Fil: Azcurra, Marcela. No especifíca;
Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
description HTLV-1 and HTLV-2 are present in different high-risk populations, such as sexual workers and injecting drug users (IDUs). HTLV-1 is endemic in areas of Middle East, Southern Japan and Latin America, whereas HTLV-2 infection is endemic among some Native Americans and some Central African tribes. The pathogenic consequences and clinical manifestations of these two viruses differ significantly, demanding an adequate identification; therefore, proper diagnosis of HTLV-1 and 2 infection is crucial. To get a final diagnosis of HTLV-1 or 2 infection, it is recommended that positive serologic samples should be confirmed by PCR assays or western blot (WB) analysis. Thus, the aim of this study was to develop and implement a simple reaction for the rapid identification of HTLV-1 and 2. Nested real-time PCR technique followed by high resolution melting was performed based on the tax/rex sequences of HTLV-1 (M2) and HTLV-2 (MoT) cell lines perfectly discriminating between HTLV-1 from HTLV-2, by distinct melting curve profiles. The sensitivity assay of this method revealed that at least 1 viral copy of HTLV-1 or 1·5 viral copy of HTLV-2 could be amplified. Later, this method was validated using 200 blood samples from corpses. In agreement with previous epidemiological, the HTLV-1 and 2 prevalence was 1·5% (CI 95%: 0·31–4·3) and 0·5% (CI 95%: 0·013–2·75), respectively. The strategy proposed herein has some advantages over other PCR-based tests because it not only reduces considerably time and the costs of the total diagnosis but also allows detection and discrimination of HTLV-1 and 2 in the same reaction.
publishDate 2022
dc.date.none.fl_str_mv 2022-10
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/188064
Caputo, Mariela; Trinks, Julieta; Azcurra, Marcela; Corach, Daniel; Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification; Wiley Blackwell Publishing, Inc; Letters in Applied Microbiology; 75; 4; 10-2022; 804-812
0266-8254
CONICET Digital
CONICET
url http://hdl.handle.net/11336/188064
identifier_str_mv Caputo, Mariela; Trinks, Julieta; Azcurra, Marcela; Corach, Daniel; Development of a nested real time PCR/high resolution melting assay for human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and 2) identification; Wiley Blackwell Publishing, Inc; Letters in Applied Microbiology; 75; 4; 10-2022; 804-812
0266-8254
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1111/lam.13752
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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