Detection of Bacteria in Cerebrospinal Fluid Using The Universal Method Based on 16S rDNA
ديما كرم عبدالمنعم ازغير, سمير برغوثي
كلية الصحة العامة-جامعة القدس · فلسطين
Meningitis is an inflammatory disease of the tissue surrounding the brain and spinal cord;the le pto-meninges. Although meningitis may be caused by viral or bacterial infection, bacterial meningitis is the most serious and notable infection of the central nervous system.It can progress rapidly with high mortality. The detection and identification of a bacterial pathogen is often essential to the physician in choosing appropriate antimicrobial the rapyand managing thein fection. The current standard for diagnosis is a microscopic examination and culturing of CSF. The definitive identification of a bacterium responsible for meningitisde pending on CSF culture should be obtained from patients presenting with clinicalsym ptoms of mening itis isessential fora successful management of mening itis .Neisseria meningitidis, Streptococcuspneumoniae, and Haemophilusinfluenzaeare fastidious organisms,andare the main causativeagents of bacterial meningitis, they may not survive long transit times or variations intemperature. As laboratories are the front line for detecting bacterial meningitis, molecular based diagnosis which is rapid and accurate, should be adapted for the detection of infectious diseases.This study represents a two-stage molecular approach for detection and identification of bacterial meningitis. The first stage depends on the application of the Universal Method for bacterial detection,the amplification of rDNA from any bacterium leading to accuratedetermination of bacterial meningitis within 3 hours of sampling.In case of negative resu lts bacterial meningitis can be ruled out. Thesam plesare simultaneously tested with multi plexcontaining specific forward primers together with ananchored general reverse primer. The multiplex contained four species-specific forward primers; its hould detect Haemo philus in fluenzae, Neisseria meningitidis ,Strep to coccus pneumoniae, and Listeria monocytogenes.Thesecan beidentified based on PCR product size. This anchoredmultiplex wasformulatedto coverthe most common bacterial agents of meningitis, in case if the multiplex failed to detect the positive cases, the Universal Method will,allowing possible identification of the pathogen as the amplified poly merase chain reaction (PCR) product is sequenced and analyzed using BLAST alignment analysis.