IMPACT OF MOLECULAR METHOD FOR THE DIAGNOSIS OF ACUTE BACTERIAL MENINGITIS IN A TERTIARY HEALTH CARE CENTRE IN NORTH INDIA
Keywords:cerebrospinal fluid (CSF), acute bacterial meningitis, PCR, MALDI-TOF
Background. Acute bacterial meningitis is one of the significant infectious diseases that add an immense burden to the health system. Proper management of meningitis is an invincible need to overcome the severe consequences.
Objectives. The aim of the study is detection of the etiological agents of acute bacterial meningitis by PCR.
Methods. Total 267 CSF samples collected from suspected bacterial meningitis cases were processed for the detection of S. pneumoniae, H. influenzae, N. meningitidis, E. coli and Group B streptococci by conventional and molecular diagnosis method. CSF was inoculated on Blood, chocolate and MacConkey agar plates and incubated at 37 °C for 24-48 hrs. Bacteria grown were identified by Matrix assisted laser desorption/ionization time of flight (MALDI-TOF). Multiplex PCR of the enlisted bacteria was performed using DNA extracted from CSF by DNA extraction kit (Qiagen, USA).
Results. 5 (1.87%) out of the total 267 CSF samples were culture positive (3 S. pneumoniae and 2 E. coli) and 28 (10.49%) had detectable DNA by conventional PCR. Out of these 28 samples, 20 (71.43%) were S. pneumoniae and 8 (28.57%) were E. coli. 15 (53.57%) out of total S. pneumoniae were present in children below 5 years of age. Similarly, E. coli was predominant in neonates. Sensitivity and specificity of the PCR was 100% and 95-98% respectively.
Conclusions. Streptococcus pneumoniae is the commonest cause of community acquired bacterial meningitis in children below five years of age. Hence, for the promising outcome, PCR should be implemented for the diagnosis of acute bacterial meningitis.
McIntyre PB, O’Brien KL, Greenwood B, Van de Beek D. Effect of vaccines on bacterial meningitis worldwide. Lancet. 2012; 380:1703–11.
Kabra SK, Praveen Kumar, Verma IC, Mukherjee D, Chowdhary BH, Sengupta S, et al. Bacterial meningitis in India: An IJP survey. Indian J Pediatr. 1991;58:505-11.
Tang LM, Chen ST, Hsu WC, Lyu RK. Acute bacterial meningitis in adults: A hospital-based epidemiological study. QJM. 1999;92:719-25.
Ayaz C, Mehmet FG, Hosoglu S, Mustafa KC, Akalin S, Omer FK. Characteristics of acute bacterial meningitis in Southeast Turkey. Indian J Med Sci. 2004;58:327-33.
Chinchankar N, Mane M, Bhave S, Bapat S, Bavdekar A, Pandit A, Niphadkar KB, Dutta A, Leboulleux D. Diagnosis and outcome of acute bacterial meningitis in early childhood. Indian pediatrics. 2002 Oct 13;39(10):914-21.
Van de Beek D, de Gans J, Tunkel AR, Wijdicks EF. Community-acquired bacterial meningitis in adults. New England Journal of Medicine. 2006 Jan 5;354(1):44-53.
Schuchat A, Robinson K, Wenger JD, Harrison LH, Farley M, Reingold AL, Lefkowitz L, Perkins BA. Bacterial meningitis in the United States in 1995. New England journal of medicine. 1997 Oct 2;337(14): 970-6.
Kasanmoentalib ES, Brouwer MC, van de Beek D. Update on bacterial meningitis: epidemiology, trials and genetic association studies. Current opinion in neurology. 2013 Jun 1;26(3):282-8.
Liu L, Oza S, Hogan D, Perin J, Rudan I, Lawn JE, Cousens S, Mathers C, Black RE. Global, regional, and national causes of child mortality in 2000–13, with projections to inform post-2015 priorities: an updated systematic analysis. The Lancet. 2015 Jan 31;385(9966):430-40.
Brouwer MC, Tunkel AR, van de Beek D. Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis. Clinical microbiology reviews. 2010 Jul 1;23(3):467-92.
Greisen K, Loeffelholz M, Purohit A, Leong D. PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid. Journal of clinical microbiology. 1994 Feb 1;32(2):335-51.
Lu JJ, Perng CL, Lee SY, Wan CC. Use of PCR with universal primers and restriction endonuclease digestions for detection and identification of common bacterial pathogens in cerebrospinal fluid. Journal of clinical microbiology. 2000 Jun 1;38(6):2076-80.
Rådström P, Bäckman A, Qian NY, Kragsbjerg P, Påhlson C, Olcén P. Detection of bacterial DNA in cerebrospinal fluid by an assay for simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and streptococci using a seminested PCR strategy. Journal of Clinical Microbiology. 1994 Nov 1;32(11):2738-44.
Saravolatz LD, Manzor O, VanderVelde N, Pawlak J, Belian B. Broad-range bacterial polymerase chain reaction for early detection of bacterial meningitis. Clinical infectious diseases. 2003 Jan 1;36(1):40-5.
Xu J, Millar BC, Moore JE, Murphy K, Webb H, Fox AJ, Cafferkey M, Crowe MJ. Employment of broad-range 16S rRNA PCR to detect aetiological agents of infection from clinical specimens in patients with acute meningitis–rapid separation of 16S rRNA PCR amplicons without the need for cloning. Journal of applied microbiology. 2003 Feb;94(2):197-206.
Schuurman T, De Boer RF, Kooistra-Smid AM, Van Zwet AA. Prospective study of use of PCR amplification and sequencing of 16S ribosomal DNA from cerebrospinal fluid for diagnosis of bacterial meningitis in a clinical setting. Journal of clinical microbiology. 2004 Feb 1;42(2):734-40.
Deutch S, Pedersen LN, Pødenphant L, Olesen R, Schmidt MB, Møller JK, Ostergaard L. Broad-range real time PCR and DNA sequencing for the diagnosis of bacterial meningitis. Scandinavian journal of infectious diseases. 2006 Jan 1;38(1):27-35.
Chakrabarti P, Das BK, Kapil A. Application of 16S rDNA based seminested PCR for diagnosis of acute bacterial meningitis. Indian Journal of Medical Research. 2009 Feb 1;129(2):182-8.
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 Journal of Medical Research. 2004 May 1;119:221-3.
Bonab ZH, Farajnia S, Ghotaslou R, Nikkhah E. Evaluation of nested PCR method for diagnosis of meningitis due to Neisseria meningitidis and Haemophilus influenzae. Turkish Journal of Biology. 2012 Nov 21;36(6):727-31.
Nigrovic LE, Malley R, Macias CG, Kanegaye JT, Moro-Sutherland DM, Schremmer RD, Schwab SH, Agrawal D, Mansour KM, Bennett JE, Katsogridakis YL. Effect of antibiotic pretreatment on cerebrospinal fluid profiles of children with bacterial meningitis. Pediatrics. 2008 Oct 1;122(4):726-30.
Nour M, Alaidarous A. Simultaneous Detection of Bacterial Meningitis in Suspected Cases of Meningitis in Children Using PCR Assay. Arch Clin Microbiol. 2017; 8:1-6.
Henry M Wu, Soraia M Cordeiro, Brian H Harcourt. BMC Infectious Diseases. 2013, 13:26.
Camacho-Gonzalez A, Spearman PW, Stoll BJ. Neonatal infectious diseases: evaluation of neonatal sepsis. Pediatric Clinics of North America. 2013 Apr;60(2):367–89.
Chang CJ, Chang WN, Huang LT, Huang SC, Chang YC, Hung PL, Lu CH, Chang CS, Cheng BC, Lee PY, Wang KW. Bacterial meningitis in infants: the epidemiology, clinical features, and prognostic factors. Brain and development. 2004 Apr 1;26(3): 168-75.
Manoharan A, Manchanda V, Balasubramanian S, Lalwani S, Modak M, Bai S, Vijayan A, Shet A, Nagaraj S, Karande S, Nataraj G. Invasive pneumococcal disease in children aged younger than 5 years in India: a surveillance study. The Lancet Infectious Diseases. 2017 Mar 1;17(3):305-12.
Jayaraman Y, Veeraraghavan B, Chethrapilly PGK. Burden of bacterial meningitis in India: Preliminary data from a hospital-based sentinel surveillance network. PLoS ONE. 2018; 13(5): e0197198.
Invasive Bacterial Infection Surveillance (IBIS) group, International Clinical Epidemiology Network (INCLEN). Prospective multicentre hospital surveillance of Streptococcus pneumoniae disease in India. Lancet. 1999; 353: 1216-21.
Sachdeva A, Kukreja S, Jain V, Dutta AK. Meningococcal disease-outbreak in Delhi. Indian Pediatrics. 2005 Jun 17;42(6):547-56.
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