A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients

Autores
Fellner, María Dolores; Durand, Karina; Correa, Mariel; Bes, David; Alonio, Lidia V.; Teyssie, Angelica R.; Picconi, María Alejandra
Año de publicación
2003
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Fil: Fellner, María Dolores. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Durand, Karina. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Correa, Mariel. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Bes, David. Hospital Nacional de Pediatría "Prof. Juan P. Garrahan"; Buenos Aires, Argentina.
Fil: Alonio, Lidia V. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Teyssié, Angélica R. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Picconi, María Alejandra. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Background: High Epstein-Barr virus load has been related to an increased risk of Posttransplant Lymphoproliferative Disorders (PTLD) in transplant recipients. Objectives: Development of a method to quantitate EBV DNA levels in peripheral blood mononuclear cells (PBMC) and evaluate its usefulness in transplant patients. Study design: We designed a semiquantitative nested PCR based on a limiting dilution analysis to detect high viral loads in PBMC. This method was applied to 25 healthy carriers, and 85 solid organ transplant recipients as follows: (A) 53 asymptomatic patients; (B) 24 symptomatic patients; (C) eight patients with PTLD. Results: In healthy carriers the reciprocal of the limiting dilution (RLD) ranged between non-detected (ND) and 1, the median RLD was ND, which is equivalent to a viral load of <1 copy per 10(5) PBMC. In the transplant population the medians RLD (range) were: (A) asymptomatic group: ND (ND-64), median equivalent to a viral load of <1 copy per 10(5) PBMC; (B) symptomatic group: 4 (ND-256), median equivalent to a range of viral load of 4-64 copies per 10(5) PBMC. (C) PTLD group: 256 (16-16384), median equivalent to a range of viral load of 256-4096 copies per 10(5) PBMC. Statistically significant differences were found between all groups: A+B vs. C (P<0.0001); A vs. B (P<0.0001); A vs. C (P<0.0001), B vs. C (P<0.0001). We also observed a good correlation between viral loads and clinical findings in four follow-up patients. Considering the RLD=256 as a cutoff point to detect transplant patients with PTLD, resulted in sensitivity 75%, specificity 96.7%, positive predictive value 60%, negative predictive value 98.3%. Conclusion: This SQ-PCR method enables us to differentiate between transplant patients with and without PTLD; therefore, it could be applied as a marker for early detection of this pathology.
Materia
Trastornos Linfoproliferativos
Valor Predictivo de las Pruebas
Trasplante de Hígado
Carga Viral
Nivel de accesibilidad
acceso abierto
Condiciones de uso
Repositorio
Sistema de Gestión del Conocimiento ANLIS MALBRÁN
Institución
Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"
OAI Identificador
oai:sgc.anlis.gob.ar:123456789/1734

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oai_identifier_str oai:sgc.anlis.gob.ar:123456789/1734
network_acronym_str SGCANLIS
repository_id_str a
network_name_str Sistema de Gestión del Conocimiento ANLIS MALBRÁN
spelling A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patientsFellner, María DoloresDurand, KarinaCorrea, MarielBes, DavidAlonio, Lidia V.Teyssie, Angelica R.Picconi, María AlejandraTrastornos LinfoproliferativosValor Predictivo de las PruebasTrasplante de HígadoCarga ViralFil: Fellner, María Dolores. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.Fil: Durand, Karina. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.Fil: Correa, Mariel. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.Fil: Bes, David. Hospital Nacional de Pediatría "Prof. Juan P. Garrahan"; Buenos Aires, Argentina.Fil: Alonio, Lidia V. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.Fil: Teyssié, Angélica R. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.Fil: Picconi, María Alejandra. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.Background: High Epstein-Barr virus load has been related to an increased risk of Posttransplant Lymphoproliferative Disorders (PTLD) in transplant recipients. Objectives: Development of a method to quantitate EBV DNA levels in peripheral blood mononuclear cells (PBMC) and evaluate its usefulness in transplant patients. Study design: We designed a semiquantitative nested PCR based on a limiting dilution analysis to detect high viral loads in PBMC. This method was applied to 25 healthy carriers, and 85 solid organ transplant recipients as follows: (A) 53 asymptomatic patients; (B) 24 symptomatic patients; (C) eight patients with PTLD. Results: In healthy carriers the reciprocal of the limiting dilution (RLD) ranged between non-detected (ND) and 1, the median RLD was ND, which is equivalent to a viral load of <1 copy per 10(5) PBMC. In the transplant population the medians RLD (range) were: (A) asymptomatic group: ND (ND-64), median equivalent to a viral load of <1 copy per 10(5) PBMC; (B) symptomatic group: 4 (ND-256), median equivalent to a range of viral load of 4-64 copies per 10(5) PBMC. (C) PTLD group: 256 (16-16384), median equivalent to a range of viral load of 256-4096 copies per 10(5) PBMC. Statistically significant differences were found between all groups: A+B vs. C (P<0.0001); A vs. B (P<0.0001); A vs. C (P<0.0001), B vs. C (P<0.0001). We also observed a good correlation between viral loads and clinical findings in four follow-up patients. Considering the RLD=256 as a cutoff point to detect transplant patients with PTLD, resulted in sensitivity 75%, specificity 96.7%, positive predictive value 60%, negative predictive value 98.3%. Conclusion: This SQ-PCR method enables us to differentiate between transplant patients with and without PTLD; therefore, it could be applied as a marker for early detection of this pathology.2003-12info:ar-repo/semantics/articuloinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdf1386-6532http://sgc.anlis.gob.ar/handle/123456789/173410.1016/s1386-6532(03)00077-5Journal of clinical virology : the official publication of the Pan American Society for Clinical Virologyenginfo:eu-repo/semantics/openAccessreponame:Sistema de Gestión del Conocimiento ANLIS MALBRÁNinstname:Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"instacron:ANLIS2025-09-29T14:30:25Zoai:sgc.anlis.gob.ar:123456789/1734Institucionalhttp://sgc.anlis.gob.ar/Organismo científico-tecnológicoNo correspondehttp://sgc.anlis.gob.ar/oai/biblioteca@anlis.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:a2025-09-29 14:30:25.913Sistema de Gestión del Conocimiento ANLIS MALBRÁN - Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"false
dc.title.none.fl_str_mv A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
title A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
spellingShingle A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
Fellner, María Dolores
Trastornos Linfoproliferativos
Valor Predictivo de las Pruebas
Trasplante de Hígado
Carga Viral
title_short A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
title_full A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
title_fullStr A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
title_full_unstemmed A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
title_sort A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients
dc.creator.none.fl_str_mv Fellner, María Dolores
Durand, Karina
Correa, Mariel
Bes, David
Alonio, Lidia V.
Teyssie, Angelica R.
Picconi, María Alejandra
author Fellner, María Dolores
author_facet Fellner, María Dolores
Durand, Karina
Correa, Mariel
Bes, David
Alonio, Lidia V.
Teyssie, Angelica R.
Picconi, María Alejandra
author_role author
author2 Durand, Karina
Correa, Mariel
Bes, David
Alonio, Lidia V.
Teyssie, Angelica R.
Picconi, María Alejandra
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Trastornos Linfoproliferativos
Valor Predictivo de las Pruebas
Trasplante de Hígado
Carga Viral
topic Trastornos Linfoproliferativos
Valor Predictivo de las Pruebas
Trasplante de Hígado
Carga Viral
dc.description.none.fl_txt_mv Fil: Fellner, María Dolores. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Durand, Karina. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Correa, Mariel. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Bes, David. Hospital Nacional de Pediatría "Prof. Juan P. Garrahan"; Buenos Aires, Argentina.
Fil: Alonio, Lidia V. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Teyssié, Angélica R. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Fil: Picconi, María Alejandra. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
Background: High Epstein-Barr virus load has been related to an increased risk of Posttransplant Lymphoproliferative Disorders (PTLD) in transplant recipients. Objectives: Development of a method to quantitate EBV DNA levels in peripheral blood mononuclear cells (PBMC) and evaluate its usefulness in transplant patients. Study design: We designed a semiquantitative nested PCR based on a limiting dilution analysis to detect high viral loads in PBMC. This method was applied to 25 healthy carriers, and 85 solid organ transplant recipients as follows: (A) 53 asymptomatic patients; (B) 24 symptomatic patients; (C) eight patients with PTLD. Results: In healthy carriers the reciprocal of the limiting dilution (RLD) ranged between non-detected (ND) and 1, the median RLD was ND, which is equivalent to a viral load of <1 copy per 10(5) PBMC. In the transplant population the medians RLD (range) were: (A) asymptomatic group: ND (ND-64), median equivalent to a viral load of <1 copy per 10(5) PBMC; (B) symptomatic group: 4 (ND-256), median equivalent to a range of viral load of 4-64 copies per 10(5) PBMC. (C) PTLD group: 256 (16-16384), median equivalent to a range of viral load of 256-4096 copies per 10(5) PBMC. Statistically significant differences were found between all groups: A+B vs. C (P<0.0001); A vs. B (P<0.0001); A vs. C (P<0.0001), B vs. C (P<0.0001). We also observed a good correlation between viral loads and clinical findings in four follow-up patients. Considering the RLD=256 as a cutoff point to detect transplant patients with PTLD, resulted in sensitivity 75%, specificity 96.7%, positive predictive value 60%, negative predictive value 98.3%. Conclusion: This SQ-PCR method enables us to differentiate between transplant patients with and without PTLD; therefore, it could be applied as a marker for early detection of this pathology.
description Fil: Fellner, María Dolores. ANLIS Dr. C. G. Malbrán. Instituto Nacional de Enfermedades Infecciosas (INEI). Servicio Virus Oncogénicos; Argentina.
publishDate 2003
dc.date.none.fl_str_mv 2003-12
dc.type.none.fl_str_mv info:ar-repo/semantics/articulo
info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv 1386-6532
http://sgc.anlis.gob.ar/handle/123456789/1734
10.1016/s1386-6532(03)00077-5
identifier_str_mv 1386-6532
10.1016/s1386-6532(03)00077-5
url http://sgc.anlis.gob.ar/handle/123456789/1734
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Sistema de Gestión del Conocimiento ANLIS MALBRÁN
instname:Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"
instacron:ANLIS
reponame_str Sistema de Gestión del Conocimiento ANLIS MALBRÁN
collection Sistema de Gestión del Conocimiento ANLIS MALBRÁN
instname_str Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"
instacron_str ANLIS
institution ANLIS
repository.name.fl_str_mv Sistema de Gestión del Conocimiento ANLIS MALBRÁN - Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"
repository.mail.fl_str_mv biblioteca@anlis.gov.ar
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