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

BackgroundIn 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.MethodBetween 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.ResultsA 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).ConclusionOur 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.

Real-time qPCR improves meningitis pathogen detection in invasive bacterial-vaccine preventable disease surveillance in Fiji

“…These results support the use of qPCR for IB-VPD surveillance, and are consistent with other published studies showing enhanced detection of these pathogens using qPCR69. The current study utilised the DNA extraction and qPCR protocols recommended by the WHO.…”

“…However, sensitivity of these methods is limited, particularly when patients receive antibiotic treatment prior to sample collection5. The use of molecular methods such as real-time quantitative PCR (qPCR) can improve sensitivity for detection of meningitis pathogens6789. The World Health Organization (WHO) recommends qPCR testing of S. pneumoniae, N. meningitidis , and H. influenzae from CSF of suspected meningitis cases as part of Invasive Bacterial Vaccine Preventable Diseases (IB-VPD) surveillance10.…”

Investigation of Bacterial Meningitis Agents from Patients with Suspicion of Non-Neonatal Meningitis by Conventional and Molecular Methods

“…pneumoniae'nın beş yaş altı çocuklarda akut bakteriyel menenjite neden olan en yaygın etken olduğu bildirilmektedir (25) . Yedi valanlı konjuge pnömokok aşısının kullanımından önce beş yaş altı çocuklarda serotip 6A, 6B, 19F ve 23F, eriş-kinlerde serotip 3 ve 23F en sık invaziv hastalık nedeni olarak bildirilirken, aşılama sonrası ise bunların yerini serotip 19A, 6C, 11A, 15A ve 15B/C almış ve menenjitli hastalarda özellikler 19A ve 22F'de artış saptanmıştır (26) .…”

“…Moleküler testler tedavi alan hastalarda dâhil olmak üzere akut bakteriyel menenjit etkeni bakterilerin tanımlamasını anlamlı bir şekilde artırmaktadır (25) . Wu ve ark.…”

“…Nhantumbo ve ark. (25) 'nın beş yaş altı 369 çocuğun BOS örneği-ni dâhil ettikleri çalışmada, PZR ile 193 (%52.3) hastada akut bakteriyel menenjit etkenlerinin belirlendiği, bu hastaların yalnızca 27 (%7.3)'sinin kültüründe üreme olduğu bildirilmiştir. Ceyhan ve ark.…”