Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries

Autores
Ducasa, Nicolás; Domínguez, Diego; Benencio, Paula; Alfie, Laura; Etcheves, Patricia; Scarton, Giampaolo; Biglione, Mirna Marcela; Caputo, Mariela
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: HTLV-1/2 exhibit a widespread distribution globally and are associated with severe clinical manifestations, necessitating precise viral identification for diagnosis. Currently, there are no official diagnostic guidelines, and a variety of published protocols exists. We introduce an enhanced nested real-time PCR technique followed by high-resolution melting (rtPCR-HRM), designed to offer a cost-effective and straightforward tool for the simultaneous identification of both viruses.Methods: The technique was tested in a retrospective, blinded study, involving a total panel of 110 samples, of which 47 were positive for HTLV-1, 12 for HTLV-2, and 51 tested negatives. Additionally, we compared the performance of this technique with a line immunoassay (LIA).Results: The results demonstrate 100 % sensitivity, specificity, and diagnostic accuracy for both viruses. Sensitivity analysis indicated that at least 1 viral copy of HTLV-1 and 14.4 viral copies of HTLV-2 could be reliably detected.Conclusions: Our results indicate that rtPCR-HRM is effective in confirming HTLV-1 and HTLV-2 infection, important in Latin American countries where both viruses circulate. Furthermore, the proposed strategy provides a new tool that can be used to resolve indeterminate cases identified by Western blot, with the added advantage of being faster and simpler than n-PCR and more cost-effective than other probe-based RT-PCRs.
Fil: Ducasa, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina
Fil: Domínguez, Diego. Hospital Zonal Caleta Olivia ; Gobierno de la Provincia de Santa Cruz;
Fil: Benencio, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina
Fil: Alfie, Laura. Universidad de Buenos Aires. Facultad de Medicina; Argentina
Fil: Etcheves, Patricia. No especifíca;
Fil: Scarton, Giampaolo. No especifíca;
Fil: Biglione, Mirna Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina
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; Argentina
Materia
HTLV-1/2
MOLECULAR DIAGNOSIS
HEALTH CARE
REAL-TIME PCR
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:11336/263326
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/263326
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countriesDucasa, NicolásDomínguez, DiegoBenencio, PaulaAlfie, LauraEtcheves, PatriciaScarton, GiampaoloBiglione, Mirna MarcelaCaputo, MarielaHTLV-1/2MOLECULAR DIAGNOSISHEALTH CAREREAL-TIME PCRhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: HTLV-1/2 exhibit a widespread distribution globally and are associated with severe clinical manifestations, necessitating precise viral identification for diagnosis. Currently, there are no official diagnostic guidelines, and a variety of published protocols exists. We introduce an enhanced nested real-time PCR technique followed by high-resolution melting (rtPCR-HRM), designed to offer a cost-effective and straightforward tool for the simultaneous identification of both viruses.Methods: The technique was tested in a retrospective, blinded study, involving a total panel of 110 samples, of which 47 were positive for HTLV-1, 12 for HTLV-2, and 51 tested negatives. Additionally, we compared the performance of this technique with a line immunoassay (LIA).Results: The results demonstrate 100 % sensitivity, specificity, and diagnostic accuracy for both viruses. Sensitivity analysis indicated that at least 1 viral copy of HTLV-1 and 14.4 viral copies of HTLV-2 could be reliably detected.Conclusions: Our results indicate that rtPCR-HRM is effective in confirming HTLV-1 and HTLV-2 infection, important in Latin American countries where both viruses circulate. Furthermore, the proposed strategy provides a new tool that can be used to resolve indeterminate cases identified by Western blot, with the added advantage of being faster and simpler than n-PCR and more cost-effective than other probe-based RT-PCRs.Fil: Ducasa, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Domínguez, Diego. Hospital Zonal Caleta Olivia ; Gobierno de la Provincia de Santa Cruz;Fil: Benencio, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Alfie, Laura. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Etcheves, Patricia. No especifíca;Fil: Scarton, Giampaolo. No especifíca;Fil: Biglione, Mirna Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier Science2024-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/263326Ducasa, Nicolás; Domínguez, Diego; Benencio, Paula; Alfie, Laura; Etcheves, Patricia; et al.; Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries; Elsevier Science; Acta Tropica; 260; 12-2024; 1-60001-706XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0001706X24002778info:eu-repo/semantics/altIdentifier/doi/10.1016/j.actatropica.2024.107395info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:01:36Zoai:ri.conicet.gov.ar:11336/263326instacron: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 10:01:37.142CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
title Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
spellingShingle Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
Ducasa, Nicolás
HTLV-1/2
MOLECULAR DIAGNOSIS
HEALTH CARE
REAL-TIME PCR
title_short Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
title_full Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
title_fullStr Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
title_full_unstemmed Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
title_sort Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries
dc.creator.none.fl_str_mv Ducasa, Nicolás
Domínguez, Diego
Benencio, Paula
Alfie, Laura
Etcheves, Patricia
Scarton, Giampaolo
Biglione, Mirna Marcela
Caputo, Mariela
author Ducasa, Nicolás
author_facet Ducasa, Nicolás
Domínguez, Diego
Benencio, Paula
Alfie, Laura
Etcheves, Patricia
Scarton, Giampaolo
Biglione, Mirna Marcela
Caputo, Mariela
author_role author
author2 Domínguez, Diego
Benencio, Paula
Alfie, Laura
Etcheves, Patricia
Scarton, Giampaolo
Biglione, Mirna Marcela
Caputo, Mariela
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv HTLV-1/2
MOLECULAR DIAGNOSIS
HEALTH CARE
REAL-TIME PCR
topic HTLV-1/2
MOLECULAR DIAGNOSIS
HEALTH CARE
REAL-TIME PCR
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Background: HTLV-1/2 exhibit a widespread distribution globally and are associated with severe clinical manifestations, necessitating precise viral identification for diagnosis. Currently, there are no official diagnostic guidelines, and a variety of published protocols exists. We introduce an enhanced nested real-time PCR technique followed by high-resolution melting (rtPCR-HRM), designed to offer a cost-effective and straightforward tool for the simultaneous identification of both viruses.Methods: The technique was tested in a retrospective, blinded study, involving a total panel of 110 samples, of which 47 were positive for HTLV-1, 12 for HTLV-2, and 51 tested negatives. Additionally, we compared the performance of this technique with a line immunoassay (LIA).Results: The results demonstrate 100 % sensitivity, specificity, and diagnostic accuracy for both viruses. Sensitivity analysis indicated that at least 1 viral copy of HTLV-1 and 14.4 viral copies of HTLV-2 could be reliably detected.Conclusions: Our results indicate that rtPCR-HRM is effective in confirming HTLV-1 and HTLV-2 infection, important in Latin American countries where both viruses circulate. Furthermore, the proposed strategy provides a new tool that can be used to resolve indeterminate cases identified by Western blot, with the added advantage of being faster and simpler than n-PCR and more cost-effective than other probe-based RT-PCRs.
Fil: Ducasa, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina
Fil: Domínguez, Diego. Hospital Zonal Caleta Olivia ; Gobierno de la Provincia de Santa Cruz;
Fil: Benencio, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina
Fil: Alfie, Laura. Universidad de Buenos Aires. Facultad de Medicina; Argentina
Fil: Etcheves, Patricia. No especifíca;
Fil: Scarton, Giampaolo. No especifíca;
Fil: Biglione, Mirna Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina
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; Argentina
description Background: HTLV-1/2 exhibit a widespread distribution globally and are associated with severe clinical manifestations, necessitating precise viral identification for diagnosis. Currently, there are no official diagnostic guidelines, and a variety of published protocols exists. We introduce an enhanced nested real-time PCR technique followed by high-resolution melting (rtPCR-HRM), designed to offer a cost-effective and straightforward tool for the simultaneous identification of both viruses.Methods: The technique was tested in a retrospective, blinded study, involving a total panel of 110 samples, of which 47 were positive for HTLV-1, 12 for HTLV-2, and 51 tested negatives. Additionally, we compared the performance of this technique with a line immunoassay (LIA).Results: The results demonstrate 100 % sensitivity, specificity, and diagnostic accuracy for both viruses. Sensitivity analysis indicated that at least 1 viral copy of HTLV-1 and 14.4 viral copies of HTLV-2 could be reliably detected.Conclusions: Our results indicate that rtPCR-HRM is effective in confirming HTLV-1 and HTLV-2 infection, important in Latin American countries where both viruses circulate. Furthermore, the proposed strategy provides a new tool that can be used to resolve indeterminate cases identified by Western blot, with the added advantage of being faster and simpler than n-PCR and more cost-effective than other probe-based RT-PCRs.
publishDate 2024
dc.date.none.fl_str_mv 2024-12
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/11336/263326
Ducasa, Nicolás; Domínguez, Diego; Benencio, Paula; Alfie, Laura; Etcheves, Patricia; et al.; Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries; Elsevier Science; Acta Tropica; 260; 12-2024; 1-6
0001-706X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/263326
identifier_str_mv Ducasa, Nicolás; Domínguez, Diego; Benencio, Paula; Alfie, Laura; Etcheves, Patricia; et al.; Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries; Elsevier Science; Acta Tropica; 260; 12-2024; 1-6
0001-706X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0001706X24002778
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.actatropica.2024.107395
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 Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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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