Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR
- Autores
- Heckel, Sofía Belén; Pacini, Antonella; Paredes, Franco; Petreli, Maria Victoria; Perez, Marilina; Adriani, Natalia; Ibarra, Guadalupe; Menzella, Hugo Gabriel; Colaneri, Alejandro Cesar; Sesma, Juliana
- Año de publicación
- 2022
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Detection of SARS-CoV-2 has created an enormous workload for laboratories worldwide resulting in a restriction at the time of massive testing. Pool testing is a strategy that reduces time and costs. However, beyond the detection of infectious diseases in blood banks, this approach is rarely implemented in routine laboratories. Therefore, what was learned from the SARS-CoV-2 pool testing should represent an opportunity to increase diagnostic capabilities. The present work, carried out in the context of a diagnostic laboratory of a public hospital during the COVID-19 pandemic, represents a contribution to this end. The main limitation of pool testing is the risk of false negatives that could have been identified by individual tests. These limitations are the dilution of samples with a low virus load during pooling and that the integrity of the sample may be affected by the quality of the sample collection. Fortunately, both limitations coincide with the main strengths of droplet digital PCR (ddPCR). ddPCR is a third-generation PCR that splits the amplification into thousands of droplets that work in parallel, increasing sensitivity and resistance to inhibitors. Therefore, ddPCR is particularly useful for pool testing. Here we show how to factor between test sensitivity and savings in test time and resources. We have identified and optimized critical parameters for pool testing. The present study, which analyzed 1000 nasopharyngeal samples, showed that the pool testing could detect even a single positive sample with a CT value of up to 30 in pools of 34 samples. This test was performed using three different standard extraction methods, the simplest being heating only, which resulted in substantial savings of extraction reagents in addition to PCR reagents. Moreover, we show that pooling can be extended to use saliva, which is less invasive and allows self-collection, reducing the risk for health personnel.
Fil: Heckel, Sofía Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; Argentina
Fil: Pacini, Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; Argentina
Fil: Paredes, Franco. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Petreli, Maria Victoria. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Perez, Marilina. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina
Fil: Adriani, Natalia. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina
Fil: Ibarra, Guadalupe. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Menzella, Hugo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Colaneri, Alejandro Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sesma, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; Argentina - Materia
-
droplet digital PCR
SARS-CoV-2
pooled samples
ddPCR
COVID-19 - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/211592
Ver los metadatos del registro completo
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Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCRHeckel, Sofía BelénPacini, AntonellaParedes, FrancoPetreli, Maria VictoriaPerez, MarilinaAdriani, NataliaIbarra, GuadalupeMenzella, Hugo GabrielColaneri, Alejandro CesarSesma, Julianadroplet digital PCRSARS-CoV-2pooled samplesddPCRCOVID-19https://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3Detection of SARS-CoV-2 has created an enormous workload for laboratories worldwide resulting in a restriction at the time of massive testing. Pool testing is a strategy that reduces time and costs. However, beyond the detection of infectious diseases in blood banks, this approach is rarely implemented in routine laboratories. Therefore, what was learned from the SARS-CoV-2 pool testing should represent an opportunity to increase diagnostic capabilities. The present work, carried out in the context of a diagnostic laboratory of a public hospital during the COVID-19 pandemic, represents a contribution to this end. The main limitation of pool testing is the risk of false negatives that could have been identified by individual tests. These limitations are the dilution of samples with a low virus load during pooling and that the integrity of the sample may be affected by the quality of the sample collection. Fortunately, both limitations coincide with the main strengths of droplet digital PCR (ddPCR). ddPCR is a third-generation PCR that splits the amplification into thousands of droplets that work in parallel, increasing sensitivity and resistance to inhibitors. Therefore, ddPCR is particularly useful for pool testing. Here we show how to factor between test sensitivity and savings in test time and resources. We have identified and optimized critical parameters for pool testing. The present study, which analyzed 1000 nasopharyngeal samples, showed that the pool testing could detect even a single positive sample with a CT value of up to 30 in pools of 34 samples. This test was performed using three different standard extraction methods, the simplest being heating only, which resulted in substantial savings of extraction reagents in addition to PCR reagents. Moreover, we show that pooling can be extended to use saliva, which is less invasive and allows self-collection, reducing the risk for health personnel.Fil: Heckel, Sofía Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; ArgentinaFil: Pacini, Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; ArgentinaFil: Paredes, Franco. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Petreli, Maria Victoria. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Perez, Marilina. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; ArgentinaFil: Adriani, Natalia. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; ArgentinaFil: Ibarra, Guadalupe. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Menzella, Hugo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Colaneri, Alejandro Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sesma, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; ArgentinaPublic Library of Science2022-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/211592Heckel, Sofía Belén; Pacini, Antonella; Paredes, Franco; Petreli, Maria Victoria; Perez, Marilina; et al.; Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR; Public Library of Science; Plos One; 17; 11-2022; 1-141932-6203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0271860info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0271860info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:44:24Zoai:ri.conicet.gov.ar:11336/211592instacron: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:34982025-09-29 10:44:24.688CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
title |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
spellingShingle |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR Heckel, Sofía Belén droplet digital PCR SARS-CoV-2 pooled samples ddPCR COVID-19 |
title_short |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
title_full |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
title_fullStr |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
title_full_unstemmed |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
title_sort |
Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR |
dc.creator.none.fl_str_mv |
Heckel, Sofía Belén Pacini, Antonella Paredes, Franco Petreli, Maria Victoria Perez, Marilina Adriani, Natalia Ibarra, Guadalupe Menzella, Hugo Gabriel Colaneri, Alejandro Cesar Sesma, Juliana |
author |
Heckel, Sofía Belén |
author_facet |
Heckel, Sofía Belén Pacini, Antonella Paredes, Franco Petreli, Maria Victoria Perez, Marilina Adriani, Natalia Ibarra, Guadalupe Menzella, Hugo Gabriel Colaneri, Alejandro Cesar Sesma, Juliana |
author_role |
author |
author2 |
Pacini, Antonella Paredes, Franco Petreli, Maria Victoria Perez, Marilina Adriani, Natalia Ibarra, Guadalupe Menzella, Hugo Gabriel Colaneri, Alejandro Cesar Sesma, Juliana |
author2_role |
author author author author author author author author author |
dc.subject.none.fl_str_mv |
droplet digital PCR SARS-CoV-2 pooled samples ddPCR COVID-19 |
topic |
droplet digital PCR SARS-CoV-2 pooled samples ddPCR COVID-19 |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/3.4 https://purl.org/becyt/ford/3 |
dc.description.none.fl_txt_mv |
Detection of SARS-CoV-2 has created an enormous workload for laboratories worldwide resulting in a restriction at the time of massive testing. Pool testing is a strategy that reduces time and costs. However, beyond the detection of infectious diseases in blood banks, this approach is rarely implemented in routine laboratories. Therefore, what was learned from the SARS-CoV-2 pool testing should represent an opportunity to increase diagnostic capabilities. The present work, carried out in the context of a diagnostic laboratory of a public hospital during the COVID-19 pandemic, represents a contribution to this end. The main limitation of pool testing is the risk of false negatives that could have been identified by individual tests. These limitations are the dilution of samples with a low virus load during pooling and that the integrity of the sample may be affected by the quality of the sample collection. Fortunately, both limitations coincide with the main strengths of droplet digital PCR (ddPCR). ddPCR is a third-generation PCR that splits the amplification into thousands of droplets that work in parallel, increasing sensitivity and resistance to inhibitors. Therefore, ddPCR is particularly useful for pool testing. Here we show how to factor between test sensitivity and savings in test time and resources. We have identified and optimized critical parameters for pool testing. The present study, which analyzed 1000 nasopharyngeal samples, showed that the pool testing could detect even a single positive sample with a CT value of up to 30 in pools of 34 samples. This test was performed using three different standard extraction methods, the simplest being heating only, which resulted in substantial savings of extraction reagents in addition to PCR reagents. Moreover, we show that pooling can be extended to use saliva, which is less invasive and allows self-collection, reducing the risk for health personnel. Fil: Heckel, Sofía Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; Argentina Fil: Pacini, Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; Argentina Fil: Paredes, Franco. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; Argentina Fil: Petreli, Maria Victoria. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; Argentina Fil: Perez, Marilina. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina Fil: Adriani, Natalia. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina Fil: Ibarra, Guadalupe. Gobierno de la Provincia de Santa Fe. Hospital Provincial de Rosario.; Argentina. Universidad Nacional de Rosario; Argentina Fil: Menzella, Hugo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina Fil: Colaneri, Alejandro Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Sesma, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Inmunología Clinica y Experimental de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología Clinica y Experimental de Rosario; Argentina |
description |
Detection of SARS-CoV-2 has created an enormous workload for laboratories worldwide resulting in a restriction at the time of massive testing. Pool testing is a strategy that reduces time and costs. However, beyond the detection of infectious diseases in blood banks, this approach is rarely implemented in routine laboratories. Therefore, what was learned from the SARS-CoV-2 pool testing should represent an opportunity to increase diagnostic capabilities. The present work, carried out in the context of a diagnostic laboratory of a public hospital during the COVID-19 pandemic, represents a contribution to this end. The main limitation of pool testing is the risk of false negatives that could have been identified by individual tests. These limitations are the dilution of samples with a low virus load during pooling and that the integrity of the sample may be affected by the quality of the sample collection. Fortunately, both limitations coincide with the main strengths of droplet digital PCR (ddPCR). ddPCR is a third-generation PCR that splits the amplification into thousands of droplets that work in parallel, increasing sensitivity and resistance to inhibitors. Therefore, ddPCR is particularly useful for pool testing. Here we show how to factor between test sensitivity and savings in test time and resources. We have identified and optimized critical parameters for pool testing. The present study, which analyzed 1000 nasopharyngeal samples, showed that the pool testing could detect even a single positive sample with a CT value of up to 30 in pools of 34 samples. This test was performed using three different standard extraction methods, the simplest being heating only, which resulted in substantial savings of extraction reagents in addition to PCR reagents. Moreover, we show that pooling can be extended to use saliva, which is less invasive and allows self-collection, reducing the risk for health personnel. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-11 |
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/11336/211592 Heckel, Sofía Belén; Pacini, Antonella; Paredes, Franco; Petreli, Maria Victoria; Perez, Marilina; et al.; Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR; Public Library of Science; Plos One; 17; 11-2022; 1-14 1932-6203 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/211592 |
identifier_str_mv |
Heckel, Sofía Belén; Pacini, Antonella; Paredes, Franco; Petreli, Maria Victoria; Perez, Marilina; et al.; Practical considerations to establish a validated platform for pooled detection of SARS-CoV-2 by droplet digital PCR; Public Library of Science; Plos One; 17; 11-2022; 1-14 1932-6203 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://journals.plos.org/plosone/article?id=10.1371/journal.pone.0271860 info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0271860 |
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Public Library of Science |
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Public Library of Science |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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