Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children

Detalhes bibliográficos
Autor(a) principal: Alenquer, M
Data de Publicação: 2022
Outros Autores: Milheiro Silva, T, Akpogheneta, O, Ferreira, F, Vale-Costa, S, Medina-Lopes, M, Batista, F, Garcia, AM, Barreto, VM, Paulino, C, Costa, J, Sobral, J, Diniz-da-Costa, M, Ladeiro, S, Corte-Real, R, Delgado Alves, J, Leite, RB, Demengeot, J, Brito, MJ, Amorim, MJ
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.17/4175
Resumo: Background: Adults are being vaccinated against SARS-CoV-2 worldwide, but the longitudinal protection of these vaccines is uncertain, given the ongoing appearance of SARS-CoV-2 variants. Children remain largely unvaccinated and are susceptible to infection, with studies reporting that they actively transmit the virus even when asymptomatic, thus affecting the community. Methods: We investigated if saliva is an effective sample for detecting SARS-CoV-2 RNA and antibodies in children, and associated viral RNA levels to infectivity. For that, we used a saliva-based SARS-CoV-2 RT-qPCR test, preceded or not by RNA extraction, in 85 children aged 10 years and under, admitted to the hospital regardless of COVID-19 symptomatology. Amongst these, 29 (63.0%) presented at least one COVID-19 symptom, 46 (54.1%) were positive for SARS-CoV-2 infection, 28 (32.9%) were under the age of 1, and the mean (SD) age was 3.8 (3.4) years. Saliva samples were collected up to 48 h after a nasopharyngeal swab-RT-qPCR test. Results: In children aged 10 years and under, the sensitivity, specificity, and accuracy of saliva-RT-qPCR tests compared to NP swab-RT-qPCR were, respectively, 84.8% (71.8%-92.4%), 100% (91.0%-100%), and 91.8% (84.0%-96.6%) with RNA extraction, and 81.8% (68.0%-90.5%), 100% (91.0%-100%), and 90.4% (82.1%-95.0%) without RNA extraction. Rescue of infectious particles from saliva was limited to CT values below 26. In addition, we found significant IgM positive responses to SARS-CoV-2 in children positive for SARS-CoV-2 by NP swab and negative by saliva compared to other groups, indicating late infection onset (>7-10 days). Conclusions: Saliva is a suitable sample type for diagnosing children aged 10 years and under, including infants aged <1 year, even bypassing RNA extraction methods. Importantly, the detected viral RNA levels were significantly above the infectivity threshold in several samples. Further investigation is required to correlate SARS-CoV-2 RNA levels to viral transmission.
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spelling Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in ChildrenSaliva molecular testingSARS-CoV-2/diagnosisChildHDE INF PEDBackground: Adults are being vaccinated against SARS-CoV-2 worldwide, but the longitudinal protection of these vaccines is uncertain, given the ongoing appearance of SARS-CoV-2 variants. Children remain largely unvaccinated and are susceptible to infection, with studies reporting that they actively transmit the virus even when asymptomatic, thus affecting the community. Methods: We investigated if saliva is an effective sample for detecting SARS-CoV-2 RNA and antibodies in children, and associated viral RNA levels to infectivity. For that, we used a saliva-based SARS-CoV-2 RT-qPCR test, preceded or not by RNA extraction, in 85 children aged 10 years and under, admitted to the hospital regardless of COVID-19 symptomatology. Amongst these, 29 (63.0%) presented at least one COVID-19 symptom, 46 (54.1%) were positive for SARS-CoV-2 infection, 28 (32.9%) were under the age of 1, and the mean (SD) age was 3.8 (3.4) years. Saliva samples were collected up to 48 h after a nasopharyngeal swab-RT-qPCR test. Results: In children aged 10 years and under, the sensitivity, specificity, and accuracy of saliva-RT-qPCR tests compared to NP swab-RT-qPCR were, respectively, 84.8% (71.8%-92.4%), 100% (91.0%-100%), and 91.8% (84.0%-96.6%) with RNA extraction, and 81.8% (68.0%-90.5%), 100% (91.0%-100%), and 90.4% (82.1%-95.0%) without RNA extraction. Rescue of infectious particles from saliva was limited to CT values below 26. In addition, we found significant IgM positive responses to SARS-CoV-2 in children positive for SARS-CoV-2 by NP swab and negative by saliva compared to other groups, indicating late infection onset (>7-10 days). Conclusions: Saliva is a suitable sample type for diagnosing children aged 10 years and under, including infants aged <1 year, even bypassing RNA extraction methods. Importantly, the detected viral RNA levels were significantly above the infectivity threshold in several samples. Further investigation is required to correlate SARS-CoV-2 RNA levels to viral transmission.Public Library of ScienceRepositório do Centro Hospitalar Universitário de Lisboa Central, EPEAlenquer, MMilheiro Silva, TAkpogheneta, OFerreira, FVale-Costa, SMedina-Lopes, MBatista, FGarcia, AMBarreto, VMPaulino, CCosta, JSobral, JDiniz-da-Costa, MLadeiro, SCorte-Real, RDelgado Alves, JLeite, RBDemengeot, JBrito, MJAmorim, MJ2022-07-29T15:03:17Z20222022-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.17/4175engPLoS One . 2022 Jun 15;17(6):e026838810.1371/journal.pone.0268388info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-03-10T09:45:39Zoai:repositorio.chlc.min-saude.pt:10400.17/4175Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T17:21:30.598379Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
title Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
spellingShingle Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
Alenquer, M
Saliva molecular testing
SARS-CoV-2/diagnosis
Child
HDE INF PED
title_short Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
title_full Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
title_fullStr Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
title_full_unstemmed Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
title_sort Saliva Molecular Testing Bypassing RNA Extraction is Suitable for Monitoring and Diagnosing SARS-CoV-2 Infection in Children
author Alenquer, M
author_facet Alenquer, M
Milheiro Silva, T
Akpogheneta, O
Ferreira, F
Vale-Costa, S
Medina-Lopes, M
Batista, F
Garcia, AM
Barreto, VM
Paulino, C
Costa, J
Sobral, J
Diniz-da-Costa, M
Ladeiro, S
Corte-Real, R
Delgado Alves, J
Leite, RB
Demengeot, J
Brito, MJ
Amorim, MJ
author_role author
author2 Milheiro Silva, T
Akpogheneta, O
Ferreira, F
Vale-Costa, S
Medina-Lopes, M
Batista, F
Garcia, AM
Barreto, VM
Paulino, C
Costa, J
Sobral, J
Diniz-da-Costa, M
Ladeiro, S
Corte-Real, R
Delgado Alves, J
Leite, RB
Demengeot, J
Brito, MJ
Amorim, MJ
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Repositório do Centro Hospitalar Universitário de Lisboa Central, EPE
dc.contributor.author.fl_str_mv Alenquer, M
Milheiro Silva, T
Akpogheneta, O
Ferreira, F
Vale-Costa, S
Medina-Lopes, M
Batista, F
Garcia, AM
Barreto, VM
Paulino, C
Costa, J
Sobral, J
Diniz-da-Costa, M
Ladeiro, S
Corte-Real, R
Delgado Alves, J
Leite, RB
Demengeot, J
Brito, MJ
Amorim, MJ
dc.subject.por.fl_str_mv Saliva molecular testing
SARS-CoV-2/diagnosis
Child
HDE INF PED
topic Saliva molecular testing
SARS-CoV-2/diagnosis
Child
HDE INF PED
description Background: Adults are being vaccinated against SARS-CoV-2 worldwide, but the longitudinal protection of these vaccines is uncertain, given the ongoing appearance of SARS-CoV-2 variants. Children remain largely unvaccinated and are susceptible to infection, with studies reporting that they actively transmit the virus even when asymptomatic, thus affecting the community. Methods: We investigated if saliva is an effective sample for detecting SARS-CoV-2 RNA and antibodies in children, and associated viral RNA levels to infectivity. For that, we used a saliva-based SARS-CoV-2 RT-qPCR test, preceded or not by RNA extraction, in 85 children aged 10 years and under, admitted to the hospital regardless of COVID-19 symptomatology. Amongst these, 29 (63.0%) presented at least one COVID-19 symptom, 46 (54.1%) were positive for SARS-CoV-2 infection, 28 (32.9%) were under the age of 1, and the mean (SD) age was 3.8 (3.4) years. Saliva samples were collected up to 48 h after a nasopharyngeal swab-RT-qPCR test. Results: In children aged 10 years and under, the sensitivity, specificity, and accuracy of saliva-RT-qPCR tests compared to NP swab-RT-qPCR were, respectively, 84.8% (71.8%-92.4%), 100% (91.0%-100%), and 91.8% (84.0%-96.6%) with RNA extraction, and 81.8% (68.0%-90.5%), 100% (91.0%-100%), and 90.4% (82.1%-95.0%) without RNA extraction. Rescue of infectious particles from saliva was limited to CT values below 26. In addition, we found significant IgM positive responses to SARS-CoV-2 in children positive for SARS-CoV-2 by NP swab and negative by saliva compared to other groups, indicating late infection onset (>7-10 days). Conclusions: Saliva is a suitable sample type for diagnosing children aged 10 years and under, including infants aged <1 year, even bypassing RNA extraction methods. Importantly, the detected viral RNA levels were significantly above the infectivity threshold in several samples. Further investigation is required to correlate SARS-CoV-2 RNA levels to viral transmission.
publishDate 2022
dc.date.none.fl_str_mv 2022-07-29T15:03:17Z
2022
2022-01-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.17/4175
url http://hdl.handle.net/10400.17/4175
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv PLoS One . 2022 Jun 15;17(6):e0268388
10.1371/journal.pone.0268388
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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