title: Use of cerebrospinal fluid and serum samples impregnated on FTATM Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae.
abstract: 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 FTATM 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 FTATM 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 FTATM 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.
  
  
  CONCLUSIONS
  The use of FTATM 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.
authors.affiliation: Center of Immunology, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Lucila Okuyama
authors.identifier: http://orcid.org/0000-0002-6688-0528
authors.initials: LO
authors.lastname: Fukasawa
authors.affiliation: Center of Immunology, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Maria Gisele
authors.initials: MG
authors.lastname: Gonçalves
authors.affiliation: Center of Immunology, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Fábio Takenori
authors.initials: FT
authors.lastname: Higa
authors.affiliation: Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Euclides Ayres
authors.initials: EA
authors.lastname: Castilho
authors.affiliation: Pan-American Health Organization, Washington DC, United States of America.
authors.forename: Ana Belén
authors.initials: AB
authors.lastname: Ibarz-Pavón
authors.affiliation: Center of Immunology, Instituto Adolfo Lutz, São Paulo, São Paulo, Brazil.
authors.forename: Claudio Tavares
authors.initials: CT
authors.lastname: Sacchi
mesh_terms: D001938:Brazil / Q000453:epidemiology; D005260:Female; D006193:Haemophilus influenzae / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D008297:Male; D016920:Meningitis, Bacterial / Q000097:blood* / Q000134:cerebrospinal fluid* / Q000453:epidemiology / Q000382:microbiology; D009345:Neisseria meningitidis / Q000302:isolation & purification* / Q000472:pathogenicity; D013296:Streptococcus pneumoniae / Q000302:isolation & purification* / Q000472:pathogenicity