Detection of Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae in Culture Negative Cerebrospinal Fluid Samples from Meningitis Patients Using a Multiplex Polymerase Chain Reaction in Nepal.

Supriya Sharma,Jyoti Acharya,Dominique A Caugant,Megha Raj Banjara,Prakash Ghimire,Anjana Singh

doi:10.3390/idr13010019

Abstract

The rapid identification of bacteria causing meningitis is crucial as delays in the treatment increase mortality rate. Though considered as the gold standard for the laboratory diagnosis of bacterial meningitis, culture might give false negative results in a case of patients under antibiotics prior to lumbar puncture. This study aimed to detect Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae by a multiplex polymerase chain reaction (PCR) in culture-negative cerebrospinal fluid samples collected from clinically suspected meningitis cases attending different hospitals in Kathmandu, Nepal from January 2017 to December 2019. S. pneumoniae, N. meningitidis and H. influenzae were detected in 8.59% (33/384) of the specimens by PCR and 7.55% (29/384) of the specimens by culture. Correlation between culture and PCR of the same sample was good (Spearman’s rho correlation coefficient = 0.932). However, the difference in positivity between culture and PCR was statistically not significant (p value > 0.05). In four specimens, culture could not detect any of the targeted bacteria whereas PCR could detect presence of H. influenzae. PCR increases the diagnostic yield for bacterial meningitis. PCR may be considered as an adjunctive test for establishing the cause of infection in culture negative clinically suspected meningitis cases.

Highlights

Bacterial meningitis is a very serious condition with death potentially occurring within few hours of the onset of symptoms, if untreated [1].The laboratory diagnosis of bacterial meningitis in low-income countries such as Nepal is based on Gram’s staining, culture and microscopic and biochemical analysis of cerebrospinal fluid (CSF) samples.Though considered to be the gold standard, CSF culture takes time and might delay diagnosis
As we aimed to compare the detection of N. meningitidis, S. pneumoniae and H. influenzae in CSF by two methods, three samples positive for E. coli were excluded in this study
To improve the diagnosis of bacterial meningitis in culture negative CSF samples, we employed multiplex polymerase chain reaction (PCR) for the detection of S. pneumoniae, N. meningitidis and H. influenzae in the CSF sample

Summary

Introduction

Bacterial meningitis is a very serious condition with death potentially occurring within few hours of the onset of symptoms, if untreated [1].The laboratory diagnosis of bacterial meningitis in low-income countries such as Nepal is based on Gram’s staining, culture and microscopic and biochemical analysis of cerebrospinal fluid (CSF) samples. Though considered to be the gold standard, CSF culture takes time and might delay diagnosis. Bacterial growth rate is low in the CSF of patients who have already received antibiotics before lumbar puncture [2]. Microscopic and biochemical analysis of CSF gives only indicative but not definitive identification of bacterial pathogens. Sensitivity of Gram’s staining of CSF varies considerably for different types of bacteria [3,4]. For effective treatment, the bacterial pathogens causing meningitis should be rapidly identified

Objectives

Methods

Discussion

Conclusion