Abstract
Purpose
Multiplex Polymerase Chain Reaction (MPCR) is a technique in which two or more gene targets are amplified in a single reaction. This has increased sensitivity of diagnosis as a single gene target may be absent in some Mycobacterium tuberculosis strains.
Methods
MPCR using two target genes specific for Mycobacterium tuberculosis, that is, IS6110 and MPB 64, ZN staining and Mycobacterial culture were performed on synovial fluid/pus samples of 80 (three confirmed, 77 suspected) patients of osteoarticular tuberculosis and 25 non tuberculosis patients.
Results
MPCR had a sensitivity of 100% in confirmed cases and 81.8% in clinically suspected cases. AFB was positive in one patient and Mycobacterial culture was positive in three patients. MPCR also had 100% specificity; MPB64 was positive in five patients in which IS6110 was negative whereas IS6110 was positive in two patients in which MPB64 was negative.
Conclusions
MPCR is a sensitive and specific method for diagnosis of paucibacilliary conditions such as osteoarticular tuberculosis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
WHO (2006) Global tuberculosis control. Surveillance, planning, financing. World Health Organization, Geneva, Switzerland, p 242
WHO (2009) The WHO's report on global TB. World Health Organization, Geneva, Switzerland
Titov AG, Vyshnevskaya EB, Mazurenko SI, Santavirta S, Konttinen YT (2004) Use of polymerase chain reaction to diagnose tuberculous arthritis from joint tissues and synovial fluid. Arch Pathol Lab Med 128:205–209
Sequeira W, Co H, Block JA (2000) Osteoarticular tuberculosis: current diagnosis and treatment. Am J Ther 7:393–398
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
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
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 Microbiol 31:2410–2416
Noel AB, Lecossier D, Nassif X, Birgite G, Frebault VL, Hance AJ (1989) Rapid diagnosis of tuberculosis by amplification of Mycobacterial DNA in clinical samples. Lancet 2:1069–1071
Narayanan S, Girish C, Mahadevan S, Rajajee S (2002) Evaluation of PCR using TRC4 and IS6110 primers in detection of tuberculous meningitis. J Clin Microbiol 39:2006–2008
Radhakrishnan I, Kumar RA, Mundayoor S (2001) Implication of low frequency of IS6110 in fingerprinting field isolates of Mycobacterium tuberculosis from Kerala. India J Clin Microbiol 39:1683
Vikas A, Shenai S, Mohrir G, Deshmukh M, Bhaduri A et al (2009) Osteoarticular tuberculosis—diagnostic solution in a disease endemic region. J Infect Dev Ctries 3:511–516
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
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
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
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
Titov AG, Vyshnevskaya EB, Mazurenko SI, Santavirta S, Konttinen YT (2004) Use of polymerase chain reaction to diagnose tuberculous arthritis from joint tissues and synovial fluid. Arch Pathol Lab Med 128:205–209
Tiwar V, Jain A, Verma RK (2003) Application of enzyme amplified mycobacterial DNA detection in the diagnosis of pulmonary and extrapulmonary tuberculosis. Indian J Med Res 118:224–228
Pandey V, Chawla K, Acharya K, Rao S, Rao S (2009) The role of polymerase chain reaction in the management of osteoarticular tuberculosis. Int Orthop 33:801–805
Noordohek GT, Kolk AH, Bjune G, Catty D, Dale JW, Fine PE et al (1994) Sensitivity and specificity of PCR for detection of Mycobacterium tuberculosis: a blind comparison study among laboratories. J Clin Microbiol 32:277–284
Bhardwaj V, Agrawal M, Suri T, Sural S, Kashyap R , Dhal A (2010) Evaluation of adequacy of short-course chemotherapy for extraspinal osteoarticular tuberculosis using 99mTc ciprofloxacin scan. Int Orthop [Epub ahead of print] Nov 30. doi:10.1007/s00264-010-1162)
Conflict of interest
None.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sharma, K., Sharma, A., Sharma, S.K. et al. Does multiplex Polymerase Chain Reaction increase the diagnostic percentage in osteoarticular tuberculosis? A prospective evaluation of 80 cases. International Orthopaedics (SICOT) 36, 255–259 (2012). https://doi.org/10.1007/s00264-011-1241-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00264-011-1241-7