Efficiency of Real-Time PCR in the Diagnosis of Community-Acquired Bacterial Meningitis in Children

Yıl 2022, Cilt: 12 Sayı: 02, 47 - 53, 15.06.2022

Shahid Raza Bimal Das Rama Chaudhry Vinay Goyal Rakesh Lodha Seema Sood Hitender Gautam Arti Kapil

https://doi.org/10.5799/jmid.1130082

Öz

Objectives: Community-acquired bacterial meningitis (CABM) is a life-threatening condition and remains a public health concern despite various efforts to prevent it. This study aimed to detect the bacteria causing CABM in children by Real-Time PCR.
Methods: In total, 178 Cerebrospinal fluid (CSF) samples from suspected meningitis cases were collected and subjected to cell count, biochemical, microbiological, and molecular analysis. Bacteria grown on blood and chocolate agar were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). DNA from CSF was extracted and used to detect bacteria by Real-Time PCR using TaqMan Probe.
Results: Fifty (28.09%) patients were diagnosed with confirmed meningitis. Of them, 46 (25.84%) were Real-Time PCR, and four (2.25%) were culture and Real-Time PCR positive. Out of 50 bacteria detected, S. pneumoniae (n=35, 19.7%) was the leading causative bacteria and was followed by H. influenzae (seven, 3.93%), E. coli (five, 2.80%), S. agalactiae (two, 1.12%), and N. meningitidis (one, 0.56%). Most of the S. pneumoniae (18 isolates, 51.4%) were isolated from 3-24 months of children, and in neonates, E. coli was the predominant bacteria. When CSF culture was the gold standard for diagnosis, the sensitivity and specificity of Real-Time PCR for S. pneumoniae were 100% (95%CI: 15.81-100%) and 81.25% (95%CI: 74.69-86.73%), respectively.
Conclusion: Streptococcus pneumoniae remains the leading organism of CABM in children despite vaccination and advancement in diagnosis. Real-time PCR has emerged as a vibrant diagnostic molecular appliance. Hence, Regular surveillance is crucial to curb the burdens and trends of CABM in children.

Anahtar Kelimeler

Streptococcus pneumoniae, Community-acquired bacterial meningitis, CSF, Real-Time PCR

Kaynakça

• 1. Van de Beek D, de Gans J, Spanjaard L, Weisfelt M, Reitsma JB, Vermeulen M. Clinical features and prognostic factors in adults with bacterial meningitis. N Engl J Med 2004; 351(28):1849-59.
• 2. Bijlsma MW, Brouwer MC, Kasanmoentalib ES. Community-acquired bacterial meningitis in adults in the Netherlands, 2006–14: a prospective cohort study. Lancet Infect Dis 2016; 16 (3): 339–347.
• 3. McIntyre PB, O’Brien KL, Greenwood B, Van de Beek D: Effect of vaccines on bacterial meningitis worldwide. Lancet 2012; 380(9854):1703-1711.
• 4. Klinger G, Chin CN, Beyene J, Perlman M: Predicting the outcome of neonatal bacterial meningitis. Pediatrics 2000;106(3): 477–482.
• 5. Stevens JP, Eames M, Kent A, Halkets S, Holt D, Harvey D. Long-term outcome of neonatal meningitis. Arch Dis Child Fetal Neonatal Ed 2003; 88(3): F179-184.
• 6. Chang CJ, Chang WN, Huang LT. Bacterial meningitis in infants: the epidemiology, clinical features, and prognostic factors. Brain Dev 2004; 26(3): 168-175.
• 7. De Louvois J, Halket S, Harvey D. Neonatal meningitis in England and Wales: sequelae at 5 years of age. Eur J Pediatr 2005;164(12): 730-734.
• 8. Brouwer MC, Tunkel AR, Van de Beek D. Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis. Clinical microbiology reviews 2010; 23 (3):467- 92.
• 9. Centres for Disease Control and Prevention (CDC) (2017) Bacterial Meningitis [updated January 25, 2017].
• 10. World Health Organization (WHO). Haemophilus influenzae type b (Hib) Vaccination Position Paper-July 2013. Releve epidemiologi quehebdomadaire.2013; 88(39): 413-426.
• 11. Hamborsky J, Kroger A, Wolfe C, editors. Centers for Disease Control and Prevention: Epidemiology and Prevention of Vaccine-Preventable Diseases. 13th ed Washington D.C.: Public Health Foundation; 2015.
• 12. Diallo AO, Soeters HM, Yameogo I, Sawadogo G, Ake F, Lingani C. Bacterial Meningitis epidemiology and return of Neisseria meningitidis serogroup A cases in Burkina Faso in the five years following MenAfriVac mass vaccination campaign. PLoS One 2017; 12(11): e0187466.
• 13. Yang S, Rothman RE. PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings. Lancet Infect Dis 2004; 4(6):337-348.
• 14. Wang Y, Guo G, Wang H, et al. Comparative study of bacteriological culture and real-time fluorescence quantitative PCR (RT-PCR) and multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay in the diagnosis of bacterial neonatal meningitis. BMC Pediatrics 2014;14: 224.
• 15. Corless CE, Guiver M, Borrow R, Edwards-Jones V, Fox AJ, Kaczmarski EB. Simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in suspected cases of meningitis and septicemia using real-time PCR. J Clin Microbiol 2001; 39(4): 1553-1558.
• 16. Bergh K, Stoelhaug A, Loeseth K, Bevanger L. Detection of group B streptococci (GBS) in vaginal swabs using real-time PCR with TaqMan probe hybridization. Indian J Med Res 2004; 119 (4): 221-223. 17. Bonab ZH, Farajania S, Ghostasloy R, Nikkah E. Evaluation of nested PCR method for diagnosis of meningitis due to Neisseria meningitidis and Haemophilus influenzae. Turk J Biol 2012; 36:727-31.
• 18. Nigrovic LE, Malley R, Macias CG, et al. Effect of antibiotic pretreatment on cerebrospinal fluid profiles of children with bacterial meningitis. Pediatrics 2008; 122 (4):726–30.
• 19. Jayaraman Y, Veeraraghavan B, Chethrapilly Purushothaman GK, Sukumar B, Kangusamy B, Nair KA. Burden of bacterial meningitis in India: Preliminary data from a hospital based sentinel surveillance network. PLoS One 2018; 13(5): e0197198.
• 20. Wu HM, Cordeiro SM, Harcourt BM, Carvalho M, Azevedo J, Oliveira TQ. Accuracy of real-time PCR, Gram stain and culture for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae meningitis diagnosis. BMC Infect Dis 2013; 13:26.
• 21. Khumalo J, Nicol M, Hardie D, Muloiwa R, Mteshana P, Bamford C. Diagnostic Accuracy of two multiplex real-time polymerase chain reaction assays for the diagnosis of meningitis in children in a resource-limited setting. PLoS One 2017;12(3): e0173948.
• 22. Oordt-Speets AM, Bolijn R, Van H RC, Bhavsar A, Kyaw MH. Global etiology of bacterial meningitis: A systematic review and meta-analysis. PLoS ONE 2018; 13(6): e0198772.
• 23. Debnath DJ, Wanjpe A, Kakrani V, Singru S. Epidemiological study of acute bacterial meningitis in admitted children below twelve years of age in a tertiary care teaching hospital in Pune, India. Med J DY Patil Univ 2012; 5: 28-30.
• 24. Manoharan A, Manchanda V, Balasubramanian S, Lalwani S, Modak M, Bai S: Invasive pneumococcal disease in children aged younger than 5 years in India: surveillance study. Lancet Infect Dis 2017; 17(3): 305-312.
• 25. Kabra SK, Praveen K, Verma IC, Mukherjee D, Chowdhary BH, Sengupta S. Bacterial meningitis in India: An IJP survey. Indian J Pediatr 1991; 58(4):505-11.
• 26. Sinclair D, Marie-Pierre P, Jacob JT, Brian G: The Epidemiology of meningococcal Disease in India. Trop Med Int Health 2010; 15(12):1421-35.
• 27. Lynda O, Christian R, Sarah K. The Epidemiology, Management, and Outcomes of Bacterial Meningitis in Infants. Pediatrics 2017; 140(1): e20170476.