title: Evaluation of multiplex PCR using MPB64 and IS6110 primers for rapid diagnosis of tuberculous meningitis.
abstract: Tuberculous meningitis (TBM) is one of those most serious manifestations of extra-pulmonary tuberculosis and prompt diagnosis and treatment is required for better clinical outcome. It is difficult to diagnose due to lack of rapid, sensitive, and specific tests. Newer methods, which are easy and reliable, are required to diagnose TBM at an early stage. Thus our aim was to evaluate the Multiplex polymerase chain reaction (PCR) technique, using primers directed against the insertion sequence IS6110 and MPB64 gene for the detection of Mycobacterium tuberculosis in Cerebrospinal fluid (CSF), for rapid diagnosis of TBM patients. 102 CSF samples were analyzed from patients suspected with TBM along with a control group of 10 patients having other neurological disorders. CSF sediments were analyzed individually for M. tuberculosis DNA by Multiplex PCR using two set of primers targeting insertion sequence IS6110 and gene MBp64, which is very specific for MTBC. Out of 37 patients diagnosed with TBM clinically, MPB64 PCR was positive in 22, IS6110 PCR was positive in 28, both PCR using Multiplex were positive in 34 and Microscopy was positive in one. Thus Sensitivity of MPB64 PCR, IS6110 PCR, Multiplex PCR and Microscopy were found to be 62.3%, 75.4%, 91.8% and 2.7% respectively. In non TBM group PCR was negative in all cases hence, the specificity was 100%. Multiplex PCR system using primers targeting IS6110 and MPB64, for the detection of M. tuberculosis DNA in CSF samples, has high sensitivity than any one of them alone, and could be used for the early detection of TBM in CSF samples.
authors.affiliation: Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Nepal; Department of Microbiology, Kathmandu Medical College and Teaching Hospital, Sinamangal, Kathmandu, Nepal. Electronic address: Lekhak.s@hotmail.com.
authors.forename: Sunil Prasad
authors.initials: SP
authors.lastname: Lekhak
authors.affiliation: Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Nepal.
authors.forename: Laxmi
authors.initials: L
authors.lastname: Sharma
authors.affiliation: Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Nepal.
authors.forename: Reema
authors.initials: R
authors.lastname: Rajbhandari
authors.affiliation: Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Nepal.
authors.forename: Pravesh
authors.initials: P
authors.lastname: Rajbhandari
authors.affiliation: Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Nepal.
authors.forename: Resha
authors.initials: R
authors.lastname: Shrestha
authors.affiliation: Annapurna Neurological Institute and Allied Sciences, Maitighar, Kathmandu, Nepal.
authors.forename: Basant
authors.initials: B
authors.lastname: Pant
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D002648:Child; D017931:DNA Primers / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000134:cerebrospinal fluid / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D014390:Tuberculosis, Meningeal / Q000175:diagnosis* / Q000235:genetics; D055815:Young Adult
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; C080865:MPB64 protein, Mycobacterium
keywords: IS6110; MPB64; Multiplex polymerase chain reaction; Mycobacterium tuberculosis; PCR; Primers; TBM; Tuberculous meningitis