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Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children

New diagnostic tests for the etiology of childhood pneumonia are needed. We evaluated the antibody-in-lymphocyte supernatant (ALS) assay to detect immunoglobulin (Ig) G secretion from ex vivo peripheral blood mononuclear cell (PBMC) culture, as a potential diagnostic test for pneumococcal pneumonia....

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Autores principales: Carter, Michael J., Gurung, Pallavi, Jones, Claire, Rajkarnikar, Shristy, Kandasamy, Rama, Gurung, Meeru, Thorson, Stephen, Gautam, Madhav C., Prajapati, Krishna G., Khadka, Bibek, Maharjan, Anju, Knight, Julian C., Murdoch, David R., Darton, Thomas C., Voysey, Merryn, Wahl, Brian, O'Brien, Katherine L., Kelly, Sarah, Ansari, Imran, Shah, Ganesh, Ekström, Nina, Melin, Merit, Pollard, Andrew J., Kelly, Dominic F., Shrestha, Shrijana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988833/
https://www.ncbi.nlm.nih.gov/pubmed/32039044
http://dx.doi.org/10.3389/fcimb.2019.00459
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author Carter, Michael J.
Gurung, Pallavi
Jones, Claire
Rajkarnikar, Shristy
Kandasamy, Rama
Gurung, Meeru
Thorson, Stephen
Gautam, Madhav C.
Prajapati, Krishna G.
Khadka, Bibek
Maharjan, Anju
Knight, Julian C.
Murdoch, David R.
Darton, Thomas C.
Voysey, Merryn
Wahl, Brian
O'Brien, Katherine L.
Kelly, Sarah
Ansari, Imran
Shah, Ganesh
Ekström, Nina
Melin, Merit
Pollard, Andrew J.
Kelly, Dominic F.
Shrestha, Shrijana
author_facet Carter, Michael J.
Gurung, Pallavi
Jones, Claire
Rajkarnikar, Shristy
Kandasamy, Rama
Gurung, Meeru
Thorson, Stephen
Gautam, Madhav C.
Prajapati, Krishna G.
Khadka, Bibek
Maharjan, Anju
Knight, Julian C.
Murdoch, David R.
Darton, Thomas C.
Voysey, Merryn
Wahl, Brian
O'Brien, Katherine L.
Kelly, Sarah
Ansari, Imran
Shah, Ganesh
Ekström, Nina
Melin, Merit
Pollard, Andrew J.
Kelly, Dominic F.
Shrestha, Shrijana
author_sort Carter, Michael J.
collection PubMed
description New diagnostic tests for the etiology of childhood pneumonia are needed. We evaluated the antibody-in-lymphocyte supernatant (ALS) assay to detect immunoglobulin (Ig) G secretion from ex vivo peripheral blood mononuclear cell (PBMC) culture, as a potential diagnostic test for pneumococcal pneumonia. We enrolled 348 children with pneumonia admitted to Patan Hospital, Kathmandu, Nepal between December 2015 and September 2016. PBMCs sampled from participants were incubated for 48 h before harvesting of cell culture supernatant (ALS). We used a fluorescence-based multiplexed immunoassay to measure the concentration of IgG in ALS against five conserved pneumococcal protein antigens. Of children with pneumonia, 68 had a confirmed etiological diagnosis: 12 children had pneumococcal pneumonia (defined as blood or pleural fluid culture-confirmed; or plasma CRP concentration ≥60 mg/l and nasopharyngeal carriage of serotype 1 pneumococci), and 56 children had non-pneumococcal pneumonia. Children with non-pneumococcal pneumonia had either a bacterial pathogen isolated from blood (six children); or C-reactive protein <60 mg/l, absence of radiographic consolidation and detection of a pathogenic virus by multiplex PCR (respiratory syncytial virus, influenza viruses, or parainfluenza viruses; 23 children). Concentrations of ALS IgG to all five pneumococcal proteins were significantly higher in children with pneumococcal pneumonia than in children with non-pneumococcal pneumonia. The concentration of IgG in ALS to the best-performing antigen discriminated between children with pneumococcal and non-pneumococcal pneumonia with a sensitivity of 1.0 (95% CI 0.73–1.0), specificity of 0.66 (95% CI 0.52–0.78) and area under the receiver-operating characteristic curve (AUROCC) 0.85 (95% CI 0.75–0.94). Children with pneumococcal pneumonia were older than children with non-pneumococcal pneumonia (median 5.6 and 2.0 years, respectively, p < 0.001). When the analysis was limited to children ≥2 years of age, assay of IgG ALS to pneumococcal proteins was unable to discriminate between children with pneumococcal pneumonia and non-pneumococcal pneumonia (AUROCC 0.67, 95% CI 0.47–0.88). This method detected spontaneous secretion of IgG to pneumococcal protein antigens from cultured PBMCs. However, when stratified by age group, assay of IgG in ALS to pneumococcal proteins showed limited utility as a test to discriminate between pneumococcal and non-pneumococcal pneumonia in children.
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spelling pubmed-69888332020-02-07 Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children Carter, Michael J. Gurung, Pallavi Jones, Claire Rajkarnikar, Shristy Kandasamy, Rama Gurung, Meeru Thorson, Stephen Gautam, Madhav C. Prajapati, Krishna G. Khadka, Bibek Maharjan, Anju Knight, Julian C. Murdoch, David R. Darton, Thomas C. Voysey, Merryn Wahl, Brian O'Brien, Katherine L. Kelly, Sarah Ansari, Imran Shah, Ganesh Ekström, Nina Melin, Merit Pollard, Andrew J. Kelly, Dominic F. Shrestha, Shrijana Front Cell Infect Microbiol Cellular and Infection Microbiology New diagnostic tests for the etiology of childhood pneumonia are needed. We evaluated the antibody-in-lymphocyte supernatant (ALS) assay to detect immunoglobulin (Ig) G secretion from ex vivo peripheral blood mononuclear cell (PBMC) culture, as a potential diagnostic test for pneumococcal pneumonia. We enrolled 348 children with pneumonia admitted to Patan Hospital, Kathmandu, Nepal between December 2015 and September 2016. PBMCs sampled from participants were incubated for 48 h before harvesting of cell culture supernatant (ALS). We used a fluorescence-based multiplexed immunoassay to measure the concentration of IgG in ALS against five conserved pneumococcal protein antigens. Of children with pneumonia, 68 had a confirmed etiological diagnosis: 12 children had pneumococcal pneumonia (defined as blood or pleural fluid culture-confirmed; or plasma CRP concentration ≥60 mg/l and nasopharyngeal carriage of serotype 1 pneumococci), and 56 children had non-pneumococcal pneumonia. Children with non-pneumococcal pneumonia had either a bacterial pathogen isolated from blood (six children); or C-reactive protein <60 mg/l, absence of radiographic consolidation and detection of a pathogenic virus by multiplex PCR (respiratory syncytial virus, influenza viruses, or parainfluenza viruses; 23 children). Concentrations of ALS IgG to all five pneumococcal proteins were significantly higher in children with pneumococcal pneumonia than in children with non-pneumococcal pneumonia. The concentration of IgG in ALS to the best-performing antigen discriminated between children with pneumococcal and non-pneumococcal pneumonia with a sensitivity of 1.0 (95% CI 0.73–1.0), specificity of 0.66 (95% CI 0.52–0.78) and area under the receiver-operating characteristic curve (AUROCC) 0.85 (95% CI 0.75–0.94). Children with pneumococcal pneumonia were older than children with non-pneumococcal pneumonia (median 5.6 and 2.0 years, respectively, p < 0.001). When the analysis was limited to children ≥2 years of age, assay of IgG ALS to pneumococcal proteins was unable to discriminate between children with pneumococcal pneumonia and non-pneumococcal pneumonia (AUROCC 0.67, 95% CI 0.47–0.88). This method detected spontaneous secretion of IgG to pneumococcal protein antigens from cultured PBMCs. However, when stratified by age group, assay of IgG in ALS to pneumococcal proteins showed limited utility as a test to discriminate between pneumococcal and non-pneumococcal pneumonia in children. Frontiers Media S.A. 2020-01-17 /pmc/articles/PMC6988833/ /pubmed/32039044 http://dx.doi.org/10.3389/fcimb.2019.00459 Text en Copyright © 2020 Carter, Gurung, Jones, Rajkarnikar, Kandasamy, Gurung, Thorson, Gautam, Prajapati, Khadka, Maharjan, Knight, Murdoch, Darton, Voysey, Wahl, O'Brien, Kelly, Ansari, Shah, Ekström, Melin, Pollard, Kelly and Shrestha. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Carter, Michael J.
Gurung, Pallavi
Jones, Claire
Rajkarnikar, Shristy
Kandasamy, Rama
Gurung, Meeru
Thorson, Stephen
Gautam, Madhav C.
Prajapati, Krishna G.
Khadka, Bibek
Maharjan, Anju
Knight, Julian C.
Murdoch, David R.
Darton, Thomas C.
Voysey, Merryn
Wahl, Brian
O'Brien, Katherine L.
Kelly, Sarah
Ansari, Imran
Shah, Ganesh
Ekström, Nina
Melin, Merit
Pollard, Andrew J.
Kelly, Dominic F.
Shrestha, Shrijana
Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children
title Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children
title_full Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children
title_fullStr Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children
title_full_unstemmed Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children
title_short Assessment of an Antibody-in-Lymphocyte Supernatant Assay for the Etiological Diagnosis of Pneumococcal Pneumonia in Children
title_sort assessment of an antibody-in-lymphocyte supernatant assay for the etiological diagnosis of pneumococcal pneumonia in children
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988833/
https://www.ncbi.nlm.nih.gov/pubmed/32039044
http://dx.doi.org/10.3389/fcimb.2019.00459
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