Cargando…
Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease
To improve the storage and transport of clinical specimens for the diagnosis of Neisseria meningitidis (Nm) infections in resource-limited settings, we have evaluated the performance of dried blood spot (DBS) and dried cerebrospinal fluid spot (DCS) assays. DBS and DCS were prepared on filter paper...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9569512/ https://www.ncbi.nlm.nih.gov/pubmed/36233182 http://dx.doi.org/10.3390/ijms231911879 |
_version_ | 1784809872358899712 |
---|---|
author | Kwambana-Adams, Brenda A. Clark, Stephen A. Tay, Nicole Agbla, Schadrac Chaguza, Chrispin Kagucia, Eunice W. Borrow, Ray Heyderman, Robert S. |
author_facet | Kwambana-Adams, Brenda A. Clark, Stephen A. Tay, Nicole Agbla, Schadrac Chaguza, Chrispin Kagucia, Eunice W. Borrow, Ray Heyderman, Robert S. |
author_sort | Kwambana-Adams, Brenda A. |
collection | PubMed |
description | To improve the storage and transport of clinical specimens for the diagnosis of Neisseria meningitidis (Nm) infections in resource-limited settings, we have evaluated the performance of dried blood spot (DBS) and dried cerebrospinal fluid spot (DCS) assays. DBS and DCS were prepared on filter paper from liquid specimens previously tested for Nm in the United Kingdom. Nm was detected and genogrouped by real-time PCR performed on crude genomic DNA extracted from the DBS (n = 226) and DCS (n = 226) specimens. Targeted whole-genome sequencing was performed on a subset of specimens, DBS (n = 4) and DCS (n = 6). The overall agreement between the analysis of liquid and dried specimens was (94.2%; 95% CI 90.8–96.7) for blood and (96.4%; 95% CI 93.5–98.0) for cerebrospinal fluid. Relative to liquid specimens as the reference, the DBS and DCS assays had sensitivities of (89.1%; 95% CI 82.7–93.8) and (94.2%; 95% CI 88.9–97.5), respectively, and both assays had specificities above 98%. A genogroup was identified by dried specimen analysis for 81.9% of the confirmed meningococcal infections. Near full-length Nm genome sequences (>86%) were obtained for all ten specimens tested which allowed determination of the sequence type, clonal complex, presence of antimicrobial resistance and other meningococcal genotyping. Dried blood and CSF filter spot assays offer a practical alternative to liquid specimens for the molecular and genomic characterisation of invasive meningococcal diseases in low-resource settings. |
format | Online Article Text |
id | pubmed-9569512 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-95695122022-10-17 Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease Kwambana-Adams, Brenda A. Clark, Stephen A. Tay, Nicole Agbla, Schadrac Chaguza, Chrispin Kagucia, Eunice W. Borrow, Ray Heyderman, Robert S. Int J Mol Sci Article To improve the storage and transport of clinical specimens for the diagnosis of Neisseria meningitidis (Nm) infections in resource-limited settings, we have evaluated the performance of dried blood spot (DBS) and dried cerebrospinal fluid spot (DCS) assays. DBS and DCS were prepared on filter paper from liquid specimens previously tested for Nm in the United Kingdom. Nm was detected and genogrouped by real-time PCR performed on crude genomic DNA extracted from the DBS (n = 226) and DCS (n = 226) specimens. Targeted whole-genome sequencing was performed on a subset of specimens, DBS (n = 4) and DCS (n = 6). The overall agreement between the analysis of liquid and dried specimens was (94.2%; 95% CI 90.8–96.7) for blood and (96.4%; 95% CI 93.5–98.0) for cerebrospinal fluid. Relative to liquid specimens as the reference, the DBS and DCS assays had sensitivities of (89.1%; 95% CI 82.7–93.8) and (94.2%; 95% CI 88.9–97.5), respectively, and both assays had specificities above 98%. A genogroup was identified by dried specimen analysis for 81.9% of the confirmed meningococcal infections. Near full-length Nm genome sequences (>86%) were obtained for all ten specimens tested which allowed determination of the sequence type, clonal complex, presence of antimicrobial resistance and other meningococcal genotyping. Dried blood and CSF filter spot assays offer a practical alternative to liquid specimens for the molecular and genomic characterisation of invasive meningococcal diseases in low-resource settings. MDPI 2022-10-06 /pmc/articles/PMC9569512/ /pubmed/36233182 http://dx.doi.org/10.3390/ijms231911879 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Kwambana-Adams, Brenda A. Clark, Stephen A. Tay, Nicole Agbla, Schadrac Chaguza, Chrispin Kagucia, Eunice W. Borrow, Ray Heyderman, Robert S. Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease |
title | Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease |
title_full | Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease |
title_fullStr | Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease |
title_full_unstemmed | Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease |
title_short | Evaluation of Dried Blood and Cerebrospinal Fluid Filter Paper Spots for Storing and Transporting Clinical Material for the Molecular Diagnosis of Invasive Meningococcal Disease |
title_sort | evaluation of dried blood and cerebrospinal fluid filter paper spots for storing and transporting clinical material for the molecular diagnosis of invasive meningococcal disease |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9569512/ https://www.ncbi.nlm.nih.gov/pubmed/36233182 http://dx.doi.org/10.3390/ijms231911879 |
work_keys_str_mv | AT kwambanaadamsbrendaa evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT clarkstephena evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT taynicole evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT agblaschadrac evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT chaguzachrispin evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT kaguciaeunicew evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT borrowray evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease AT heydermanroberts evaluationofdriedbloodandcerebrospinalfluidfilterpaperspotsforstoringandtransportingclinicalmaterialforthemoleculardiagnosisofinvasivemeningococcaldisease |