Abstract
Rapid and specific diagnosis of tubercular meningitis is of paramount importance to decrease morbidity and mortality. The aim of the study was to evaluate multiplex PCR using protein b, MPB 64, and IS6110 primers directed against M. tuberculosis complex for the diagnosis of tuberculous meningitis (TBM). Multiplex PCR was performed on 18 TBM confirmed cases (culture was positive), 92 clinically suspected TBM cases and 100 non-TBM (control group) patients. Multiplex PCR had a sensitivity of 94.4% for confirmed cases and specificity of 100% for confirmed TBM cases. In 92 clinically diagnosed but unconfirmed TBM cases, multiplex PCR was positive in 84.78% cases. The overall sensitivity of microscopy, culture and multiplex cases were 1.81, 16.73, and 86.63% and specificity was 100, 100, and 100% respectively. Multiplex PCR using protein b, MPB 64, and IS6110 primers has a high sensitivity and specificity in diagnosis of tubercular meningitis.
Similar content being viewed by others
References
Kulkarni SP, Jaleel MA, Kadival GV (2005) Evaluation of an in-house PCR for diagnosis of tuberculous meningitis in Indian children. J Med Microbiol 34:369–373
Bonington A, Strang JI, Klapper PE, Hood SV, Parish A, Swift PJ, Damba J, Stevens H et al (2000) TB PCR in early diagnosis of tuberculous meningitis: evaluation of Roche semi-automated COBAS Amplicor MTB test with reference to manual Amplicor MTB PCR test. Tuber Lung Dis 80:191–196
Hosoglu S, Ayaz C, Geylk MF, Kokoglu OF, Cerviz A (1998) Tuberculosis meningitis in adults: an 11 year review. Int J Tuberc Lung Dis 2:553–557
Tiwari V, Jain A, Verma RK (2003) Application of enzyme amplified mycobacterial DNA detection in the diagnosis of pulmonary & extra-pulmonary tuberculosis. Indian J Med Res 118:224–228
Clarridge JE, Shawar RM, Shinnick TM, Plikaytis BB (1993) Large scale use of polymerase chain reaction for detection of Mycobacterium tuberculosis in a routine Mycobacteriology laboratory. J Clin Microbiol 3:2049–2056
Kolk AH, Schuitema AR, Kuijper S, Van Leeuwen J, Harmans PW, Van Embden JD et al (1992) Detection of Mycobacterium tuberculosis in clinical samples by using PCR and a non-reactive detection system. J Clin Microbiol 30:2567–2575
Pao CC, Yen TS, You JB, Maa JS, Fiss EH, Chang CH (1990) Detection and identification of Mycobacterium tuberculosis by DNA amplification. J Clin Microbiol 28:1877–1890
Pai M, Flores LL, Pai N, Hubbard A, Riley LM, Colford JM (2003) Diagnostic accuracy of nucleic acid amplification tests for tuberculous meningitis: a systematic review and meta-analysis. Lancet infect Dis 3:633–643
Hooker JA, Muhindi DW, Amayo EO, Mcligeyo SO, Mand Bhatt K, Odhiambo JA (2003) A diagnostic utility of cerebrospinal fluid studies in patients with clinically suspected tuberculous meningitis. Int J Tuberc Lung Dis 7:787–796
Bhigjee AI, Padayachee R, Paruk H, Hallwith-Pillay KD, Maris S, Connoly C (2007) Diagnosis of tuberculous meningitis: clinical and laboratory parameters. Int J Infect Dis 11:348–354
Jonas V, Alden MJ, Curry JI, Kamisango K, Knott CA, Lankford R, Wolfe JM, Moore DF (1993) Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by amplification of rRNA. J Clin Microbio 31:2410–2416
Noel AB, Lecossier D, Nassif X, Gicquel Birgite, Frebault VL, Hance AJ (1989) Rapid diagnosis of tuberculosis by amplification of Mycobacterial DNA in clinical samples. Lancet 2(8671):1069–1071
Chauhan DS, Sharma VD, Prashar D, Chauhan A, Singh HB et al (2007) Molecular typing of Mycobacterial tuberculosis isolates from different parts of India based on IS6110 element polymorphism using RFLP analysis. Indian J Med Res 125:577–581
Das S, Paramasivan CN, Lowrie DB, Prabhakar R, Narayaan PR (1995) IS6110 restriction fragment length polymorphism typing of clinical isolates of Mycobacterium tuberculosis from patients with pulmonary TB in Madras, South India. Tuberc Lung Dis 76:550–554
Afrose D, Mir AW, Kirmani A, Rhehman S, Eachkoti R, Siddiqi MA (2008) Improved diagnosis of centeral nervous system tuberculosis by Mpb64-target PCR. Brazilian J Microbiol 39:209–213
Sharma K, Sharma A, Singh M, Ray P, Dandora R, Sharma SK, Modi M, Prabhakar S, Sharma M (2010) Evaluation of PCR using Protein b primers for rapid diagnosis of tuberculous meningitis. Neurol India 58:727–731
Rafi A, Naghily B (2003) Efficiency of polymerase chain reaction for the diagnosis of tuberculous meningitis. Southeast Asian J Trop Med Public Health 32:357–360
Nguyen LN, Kox LF, Pham LD, Kuijper S, Kolk AH (1996) The potential contribution of the polymerase chain reaction to the diagnosis of tuberculous meningitis. Arch Neural 53:771–776
Ahuja GK, Mohan KK, Prasad K, Behari M (1996) Diagnostic criteria for tuberculous meningitis and their validation. Tuber Lung Dis 75:149–152
Donald PR, Victor TC, Jordaan AM, Schoeman JF, Van Heldon PD (1993) Polymerase chain reaction in the diagnosis of tuberculous meningitis. Scand J Infect Dis 25:613–617
Rafi W, Venkataswamy MM, Ravi V, Chandramuki A (2007) Rapid diagnosis of tuberculous meningitis: a comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, MPB64 and 65 KDa antigen. J Neurol Sci 252:163–168
Halder S, Sharma N, Gupta VK, Tyagi JS (2009) Efficient diagnosis of tuberculous meningitis by detection of Mycobacterium tuberculosis DNA in cerebrospinal fluid filtrates using PCR. J Med Microbiol 58:616–624
Montoya JC, Recuisado G, Somberol T (1997) Detection of Mycobacterium tuberculosis cerebrospinal fluid of smear negative tuberculous meningitis using polymerase chain reaction with nested amplification. Phil J Microbiol Infect Dis 26:1–4
Seth P, Ahuja GK, Bhanu NV, Behari M, Bhowmik S, Broor S et al (1996) Evaluation of polymerase chain reaction for rapid diagnosis of clinically suspected tuberculous meningitis. Tuber Lung Dis 77:353–357
Caws M, Wison SM, Ciough C, Drodniewski F (2000) Role of IS6110 targated PCR, culture, biochemical, clinical and immunological criteria for diagnosis of tuberculous meningitis. J Clin Microbiol 38:3150–3155
Sumi MG, Mathai A, Reuben S, Sarda C, Radhakrishanan W (2002) A comparative evaluation of DOT immunobinding assay (Dot-lba) and polymerase chain reaction for the laboratory diagnosis of tuberculosis meningitis. Diagn Microbial infect Dis 35:35–38
Noordohek GT, Kolk AH, Bjune G, Catty D, Dale JW, Fine PE et al (1994) Senstivity and specificity of PCR for detection of Mycobacterium tuberculosis: a blind comparison study among laboratories. J Clin Microbiol 32:277–284
Michael JS, Lalitha MK, Cherian T, Thomas K, Mathai D, Abraham OC, Brahmadathan KN (2002) Evaluation of polymerase chain reaction for rapid diagnosis of tuberculous meningitis. Ind J Tub 49:133–137
Narayanan S, Parandaman V, Narayanan PR, Venkatesan P, Girish C, Mahadevan S, Rajajee S (2001) Evaluation of PCR using TRC(4) and IS6110 primers in detection of tuberculous meningitis. J Clin Microbiol 39:2006–2008
Conflict of interest
None.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kusum, S., Aman, S., Pallab, R. et al. Multiplex PCR for rapid diagnosis of tuberculous meningitis. J Neurol 258, 1781–1787 (2011). https://doi.org/10.1007/s00415-011-6010-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00415-011-6010-4