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Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae

BACKGROUND: The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of exis...

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Autores principales: Fukasawa, Lucila Okuyama, Gonçalves, Maria Gisele, Higa, Fábio Takenori, Castilho, Euclides Ayres, Ibarz-Pavón, Ana Belén, Sacchi, Claudio Tavares
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5325563/
https://www.ncbi.nlm.nih.gov/pubmed/28235065
http://dx.doi.org/10.1371/journal.pone.0172794
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author Fukasawa, Lucila Okuyama
Gonçalves, Maria Gisele
Higa, Fábio Takenori
Castilho, Euclides Ayres
Ibarz-Pavón, Ana Belén
Sacchi, Claudio Tavares
author_facet Fukasawa, Lucila Okuyama
Gonçalves, Maria Gisele
Higa, Fábio Takenori
Castilho, Euclides Ayres
Ibarz-Pavón, Ana Belén
Sacchi, Claudio Tavares
author_sort Fukasawa, Lucila Okuyama
collection PubMed
description BACKGROUND: The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTA(TM) Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo’s national surveillance for routine diagnosis were selected for this study. METHODS: The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory’s standard DNA extraction, and processing the same samples after being dried and stored on FTA(TM) card, and DNA extracted following the manufacturer’s instructions. RESULTS: The sensitivities for detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from CSF dried and stored on FTA(TM) cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory’s standard methodology, results showed high concordance, with Kappa index ranges of 0.9877–1.00 for CSF, and 0.8004–1.00 for serum samples. CONCLUSION: The use of FTA(TM) cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost.
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spelling pubmed-53255632017-03-09 Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae Fukasawa, Lucila Okuyama Gonçalves, Maria Gisele Higa, Fábio Takenori Castilho, Euclides Ayres Ibarz-Pavón, Ana Belén Sacchi, Claudio Tavares PLoS One Research Article BACKGROUND: The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTA(TM) Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo’s national surveillance for routine diagnosis were selected for this study. METHODS: The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory’s standard DNA extraction, and processing the same samples after being dried and stored on FTA(TM) card, and DNA extracted following the manufacturer’s instructions. RESULTS: The sensitivities for detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from CSF dried and stored on FTA(TM) cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory’s standard methodology, results showed high concordance, with Kappa index ranges of 0.9877–1.00 for CSF, and 0.8004–1.00 for serum samples. CONCLUSION: The use of FTA(TM) cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost. Public Library of Science 2017-02-24 /pmc/articles/PMC5325563/ /pubmed/28235065 http://dx.doi.org/10.1371/journal.pone.0172794 Text en © 2017 Fukasawa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Fukasawa, Lucila Okuyama
Gonçalves, Maria Gisele
Higa, Fábio Takenori
Castilho, Euclides Ayres
Ibarz-Pavón, Ana Belén
Sacchi, Claudio Tavares
Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae
title Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae
title_full Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae
title_fullStr Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae
title_full_unstemmed Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae
title_short Use of cerebrospinal fluid and serum samples impregnated on FTA(TM) Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae
title_sort use of cerebrospinal fluid and serum samples impregnated on fta(tm) elute filter paper for the diagnosis of infections caused by neisseria meningitidis, streptococcus pneumoniae and haemophilus influenzae
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5325563/
https://www.ncbi.nlm.nih.gov/pubmed/28235065
http://dx.doi.org/10.1371/journal.pone.0172794
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