Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs

Autores
Gainotti, Rosana; Ricarte, Joaquina Carmen; Ebekian, Beatriz Virginia; Videla, Cristina Mónica; Carballal, Guadalupe; Damonte, Elsa Beatriz; Echavarría, Marcela Silvia
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Human adenoviruses (HAdV) are associated with respiratory, ocular and gastrointestinal infections as well as potentially fatal disseminated disease in highly immunocompromised patients. Although there is no specific FDA approved treatment for HAdV infections, some antivirals are used in certain patients. The in vitro antiviral assays for HAdV are not standardized and are usually time consuming. The objective of this study was to evaluate a real time PCR assay for rapid determination of susceptibility of HAdV to antiviral drugs. The nucleoside analogue stavudine (d4T) was used as test drug in A549 cells infected with HAdV5. The antiviral assay measured the reduction of the HAdV DNA levels in culture supernatants by real time PCR using specific primers that amplify a conserved region of the hexon gene. This real time PCR assay demonstrated that stavudine was a selective inhibitor for HAdV5, since the effective concentration 50% (EC50) ranged from 0.08 to 0.12 mM at multiplicity of infection between 0.001 and 1. Furthermore, EC50 showed a high correlation with plaque reduction and virus yield inhibition assays (r2 = 0.9938 and r2 = 0.9468, respectively). In conclusion, the real time PCR-based antiviral assay is rapid, reproducible and could replace classical and more labor-intensive infectivity assays.
Fil: Gainotti, Rosana. Centro de Educación Médica e Investigaciones Clínicas "Norberto Quirno"; Argentina
Fil: Ricarte, Joaquina Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Ebekian, Beatriz Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Videla, Cristina Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Carballal, Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Damonte, Elsa Beatriz. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Echavarría, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Materia
ADENOVIRUS
ANTIVIRAL DRUG
REAL TIME PCR
STAVUDINE
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:11336/188251
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/188251
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugsGainotti, RosanaRicarte, Joaquina CarmenEbekian, Beatriz VirginiaVidela, Cristina MónicaCarballal, GuadalupeDamonte, Elsa BeatrizEchavarría, Marcela SilviaADENOVIRUSANTIVIRAL DRUGREAL TIME PCRSTAVUDINEhttps://purl.org/becyt/ford/3.2https://purl.org/becyt/ford/3Human adenoviruses (HAdV) are associated with respiratory, ocular and gastrointestinal infections as well as potentially fatal disseminated disease in highly immunocompromised patients. Although there is no specific FDA approved treatment for HAdV infections, some antivirals are used in certain patients. The in vitro antiviral assays for HAdV are not standardized and are usually time consuming. The objective of this study was to evaluate a real time PCR assay for rapid determination of susceptibility of HAdV to antiviral drugs. The nucleoside analogue stavudine (d4T) was used as test drug in A549 cells infected with HAdV5. The antiviral assay measured the reduction of the HAdV DNA levels in culture supernatants by real time PCR using specific primers that amplify a conserved region of the hexon gene. This real time PCR assay demonstrated that stavudine was a selective inhibitor for HAdV5, since the effective concentration 50% (EC50) ranged from 0.08 to 0.12 mM at multiplicity of infection between 0.001 and 1. Furthermore, EC50 showed a high correlation with plaque reduction and virus yield inhibition assays (r2 = 0.9938 and r2 = 0.9468, respectively). In conclusion, the real time PCR-based antiviral assay is rapid, reproducible and could replace classical and more labor-intensive infectivity assays.Fil: Gainotti, Rosana. Centro de Educación Médica e Investigaciones Clínicas "Norberto Quirno"; ArgentinaFil: Ricarte, Joaquina Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Ebekian, Beatriz Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Videla, Cristina Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Carballal, Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Damonte, Elsa Beatriz. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Echavarría, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaElsevier Science2010-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/188251Gainotti, Rosana; Ricarte, Joaquina Carmen; Ebekian, Beatriz Virginia; Videla, Cristina Mónica; Carballal, Guadalupe; et al.; Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs; Elsevier Science; Journal of Virological Methods; 164; 1-2; 3-2010; 30-340166-0934CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0166093409005047info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jviromet.2009.11.020info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:09:09Zoai:ri.conicet.gov.ar:11336/188251instacron: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:34982025-09-10 13:09:10.001CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
title Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
spellingShingle Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
Gainotti, Rosana
ADENOVIRUS
ANTIVIRAL DRUG
REAL TIME PCR
STAVUDINE
title_short Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
title_full Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
title_fullStr Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
title_full_unstemmed Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
title_sort Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs
dc.creator.none.fl_str_mv Gainotti, Rosana
Ricarte, Joaquina Carmen
Ebekian, Beatriz Virginia
Videla, Cristina Mónica
Carballal, Guadalupe
Damonte, Elsa Beatriz
Echavarría, Marcela Silvia
author Gainotti, Rosana
author_facet Gainotti, Rosana
Ricarte, Joaquina Carmen
Ebekian, Beatriz Virginia
Videla, Cristina Mónica
Carballal, Guadalupe
Damonte, Elsa Beatriz
Echavarría, Marcela Silvia
author_role author
author2 Ricarte, Joaquina Carmen
Ebekian, Beatriz Virginia
Videla, Cristina Mónica
Carballal, Guadalupe
Damonte, Elsa Beatriz
Echavarría, Marcela Silvia
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv ADENOVIRUS
ANTIVIRAL DRUG
REAL TIME PCR
STAVUDINE
topic ADENOVIRUS
ANTIVIRAL DRUG
REAL TIME PCR
STAVUDINE
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.2
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Human adenoviruses (HAdV) are associated with respiratory, ocular and gastrointestinal infections as well as potentially fatal disseminated disease in highly immunocompromised patients. Although there is no specific FDA approved treatment for HAdV infections, some antivirals are used in certain patients. The in vitro antiviral assays for HAdV are not standardized and are usually time consuming. The objective of this study was to evaluate a real time PCR assay for rapid determination of susceptibility of HAdV to antiviral drugs. The nucleoside analogue stavudine (d4T) was used as test drug in A549 cells infected with HAdV5. The antiviral assay measured the reduction of the HAdV DNA levels in culture supernatants by real time PCR using specific primers that amplify a conserved region of the hexon gene. This real time PCR assay demonstrated that stavudine was a selective inhibitor for HAdV5, since the effective concentration 50% (EC50) ranged from 0.08 to 0.12 mM at multiplicity of infection between 0.001 and 1. Furthermore, EC50 showed a high correlation with plaque reduction and virus yield inhibition assays (r2 = 0.9938 and r2 = 0.9468, respectively). In conclusion, the real time PCR-based antiviral assay is rapid, reproducible and could replace classical and more labor-intensive infectivity assays.
Fil: Gainotti, Rosana. Centro de Educación Médica e Investigaciones Clínicas "Norberto Quirno"; Argentina
Fil: Ricarte, Joaquina Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Ebekian, Beatriz Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Videla, Cristina Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Carballal, Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
Fil: Damonte, Elsa Beatriz. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Echavarría, Marcela Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; Argentina
description Human adenoviruses (HAdV) are associated with respiratory, ocular and gastrointestinal infections as well as potentially fatal disseminated disease in highly immunocompromised patients. Although there is no specific FDA approved treatment for HAdV infections, some antivirals are used in certain patients. The in vitro antiviral assays for HAdV are not standardized and are usually time consuming. The objective of this study was to evaluate a real time PCR assay for rapid determination of susceptibility of HAdV to antiviral drugs. The nucleoside analogue stavudine (d4T) was used as test drug in A549 cells infected with HAdV5. The antiviral assay measured the reduction of the HAdV DNA levels in culture supernatants by real time PCR using specific primers that amplify a conserved region of the hexon gene. This real time PCR assay demonstrated that stavudine was a selective inhibitor for HAdV5, since the effective concentration 50% (EC50) ranged from 0.08 to 0.12 mM at multiplicity of infection between 0.001 and 1. Furthermore, EC50 showed a high correlation with plaque reduction and virus yield inhibition assays (r2 = 0.9938 and r2 = 0.9468, respectively). In conclusion, the real time PCR-based antiviral assay is rapid, reproducible and could replace classical and more labor-intensive infectivity assays.
publishDate 2010
dc.date.none.fl_str_mv 2010-03
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/11336/188251
Gainotti, Rosana; Ricarte, Joaquina Carmen; Ebekian, Beatriz Virginia; Videla, Cristina Mónica; Carballal, Guadalupe; et al.; Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs; Elsevier Science; Journal of Virological Methods; 164; 1-2; 3-2010; 30-34
0166-0934
CONICET Digital
CONICET
url http://hdl.handle.net/11336/188251
identifier_str_mv Gainotti, Rosana; Ricarte, Joaquina Carmen; Ebekian, Beatriz Virginia; Videla, Cristina Mónica; Carballal, Guadalupe; et al.; Real time PCR for rapid determination of susceptibility of adenovirus to antiviral drugs; Elsevier Science; Journal of Virological Methods; 164; 1-2; 3-2010; 30-34
0166-0934
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://www.sciencedirect.com/science/article/pii/S0166093409005047
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jviromet.2009.11.020
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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score 12.993085

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