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. 2015 Sep 22;10(9):e0138249.
doi: 10.1371/journal.pone.0138249. eCollection 2015.

Frequency of Pathogenic Paediatric Bacterial Meningitis in Mozambique: The Critical Role of Multiplex Real-Time Polymerase Chain Reaction to Estimate the Burden of Disease

Aquino Albino Nhantumbo  1 , Vlademir Vicente Cantarelli  2 , Juliana Caireão  3 , Alcides Moniz Munguambe  1 , Charlotte Elizabeth Comé  1 , Gabriela do Carmo Pinto  4 , Tomás Francisco Zimba  5 , Inácio Mandomando  6 , Cynthia Baltazar Semá  7 , Cícero Dias  3 , Milton Ozório Moraes  8 , Eduardo Samo Gudo  7
Affiliations

Frequency of Pathogenic Paediatric Bacterial Meningitis in Mozambique: The Critical Role of Multiplex Real-Time Polymerase Chain Reaction to Estimate the Burden of Disease

Aquino Albino Nhantumbo et al. PLoS One. .

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.

Method: 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).

Conclusion: 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.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Numbers of patients enrolled and tested.
The flow chart show the number of children under 5 years old who were admitted at each hospital during the study period. ABM: acute bacterial meningitis; CSF: Cerebrospinal fluid; HCB: Hospital Central da Beira; HCM: Hospital Central de Maputo; HCN: Hospital Central de Nampula; M-qPCR: multiplex real-time polymerase chain reaction; NMRL: National Reference Microbiology Laboratory
Fig 2
Fig 2. Proportion of pathogens causing acute bacterial meningitis cases in Mozambique in the period between March 2013 and March 2014.
This figure show the proportion of pathogens causing ABM in Mozambique in the period between March 2013 and March 2014 using multiplex real-time polymerase chain reaction (M-qPCR). ABM: Acute Bacterial Meningitis.
Fig 3
Fig 3. Monthly variation of the relative frequency of pathogens causing ABM.
This figure show the monthly variation of relative frequency of pathogens causing ABM and also the variation in the number of CSF samples collected from children <5 years. Frequency of pathogens was determined using multiplex real-time polymerase chain reaction (M-qPCR). ABM: acute bacterial meningitis; CSF: cerebrospinal fluid
See this image and copyright information in PMC

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