title: Frequency of Pathogenic Paediatric Bacterial Meningitis in Mozambique: The Critical Role of Multiplex Real-Time Polymerase Chain Reaction to Estimate the Burden of Disease.
abstract: BACKGROUND
  In Sub-Saharan Africa, including Mozambique, acute bacterial meningitis (ABM) represents a main cause of childhood mortality. The burden of ABM is seriously underestimated because of the poor performance of culture sampling, the primary method of ABM surveillance in the region. Low quality cerebrospinal fluid (CSF) samples and frequent consumption of antibiotics prior to sample collection lead to a high rate of false-negative results. To our knowledge, this study is the first to determine the frequency of ABM in Mozambique using real-time polymerase chain reaction (qPCR) and to compare results to those of culture sampling.
  
  
  METHODS
  Between March 2013 and March 2014, CSF samples were collected at 3 regional hospitals from patients under 5 years of age, who met World Health Organization case definition criteria for ABM. Macroscopic examination, cytochemical study, culture, and qPCR were performed on all samples.
  
  
  RESULTS
  A total of 369 CSF samples were collected from children clinically suspected of ABM. qPCR showed a significantly higher detection rate of ABM-causing pathogens when compared to culture (52.3% [193/369] versus 7.3% [27/369], p = 0.000). The frequency of Streptococcus pneumoniae, Haemophilus influenzae, group B Streptococci, and Neisseria meningitidis were 32.8% (121⁄369), 12.2%, (45⁄369), 3.0% (16⁄369) and 4.3% (11⁄369), respectively, significantly higher compared to that obtained on culture (p < 0.001 for each).
  
  
  CONCLUSIONS
  Our findings demonstrate that culture is less effective for the diagnosis of ABM than qPCR. The common use of culture rather than qPCR to identify ABM results in serious underestimation of the burden of the disease, and our findings strongly suggest that qPCR should be incorporated into surveillance activities for ABM. In addition, our data showed that S. pneumoniae represents the most common cause of ABM in children under 5 years of age.
authors.affiliation: Laboratório Nacional de Referência de Microbiologia, Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Aquino Albino
authors.initials: AA
authors.lastname: Nhantumbo
authors.affiliation: Universidade Feevale, Rio Sul, Brazil; Universidade Federal de Ciências de Saúde de Porto Algre (UFCSPA), Porto Alegre, Brazil.
authors.forename: Vlademir Vicente
authors.initials: VV
authors.lastname: Cantarelli
authors.affiliation: Universidade Federal de Ciências de Saúde de Porto Algre (UFCSPA), Porto Alegre, Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: Caireão
authors.affiliation: Laboratório Nacional de Referência de Microbiologia, Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Alcides Moniz
authors.initials: AM
authors.lastname: Munguambe
authors.affiliation: Laboratório Nacional de Referência de Microbiologia, Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Charlotte Elizabeth
authors.initials: CE
authors.lastname: Comé
authors.affiliation: Laboratório de Isolamento Viral, Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Gabriela do Carmo
authors.initials: Gdo C
authors.lastname: Pinto
authors.affiliation: Departamento de Medicina at the Hospital Central de Maputo, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Tomás Francisco
authors.initials: TF
authors.lastname: Zimba
authors.affiliation: Centro de Investigação em Saúde da Manhiça, Ministério de Saúde, Maputo, Mozambique.
authors.forename: Inácio
authors.initials: I
authors.lastname: Mandomando
authors.affiliation: Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Cynthia Baltazar
authors.initials: CB
authors.lastname: Semá
authors.affiliation: Universidade Federal de Ciências de Saúde de Porto Algre (UFCSPA), Porto Alegre, Brazil.
authors.forename: Cícero
authors.initials: C
authors.lastname: Dias
authors.affiliation: Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Milton Ozório
authors.initials: MO
authors.lastname: Moraes
authors.affiliation: Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique.
authors.forename: Eduardo Samo
authors.initials: ES
authors.lastname: Gudo
mesh_terms: D002675:Child, Preschool; D004269:DNA, Bacterial / Q000134:cerebrospinal fluid* / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005260:Female; D006193:Haemophilus influenzae / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D016015:Logistic Models; D008297:Male; D016920:Meningitis, Bacterial / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology*; D008826:Microbial Sensitivity Tests; D009073:Mozambique / Q000453:epidemiology; D015999:Multivariate Analysis; D009345:Neisseria meningitidis / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012621:Seasons; D012680:Sensitivity and Specificity; D012720:Severity of Illness Index; D013292:Streptococcus agalactiae / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial