Does multiplex Polymerase Chain Reaction increase the diagnostic percentage in osteoarticular tuberculosis? A prospective evaluation of 80 cases

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.

Multiplex polymerase chain reaction using insertion sequence 6110 (IS6110) and mycobacterial protein fraction from BCG of Rm 0.64 in electrophoresis target genes for diagnosis of tuberculous lymphadenitis

“…We had also reported previously that multiplex targets increase the sensitivity of the tests especially in smear-negative extrapulmonary conditions such as tubercular meningitis and osteoarticular tuberculosis. [7,8] MPCR has many advantages such as less chances of contamination as all reaction takes place in same tube and cost-effective, as stated earlier. [7,8] Therefore, in the present study we evaluated MPCR targeting IS6110 and MPB64 together for diagnosis of the M. tuberculosis complex in clinically suspected TBLA cases.…”

“…[7,8,12] Multiplex polymerase chain reaction MPCR was performed using primers (Sigma, Bangalore, India) for the IS6110 (IS1: 5'-CCTGCGAGCGTAGGCGT-3' and IS2: 5'-CTCGTCCA GCGCCGCTTCGG-3') and MPB64 (MPB1: 5'-ACC AGGGAGCGGTTCGCCTGG-3' and MPB2: 5'-GATCT GGGGGTCGTCGGAGCT-3') region of the M. tuberculosis complex; amplifi cation conditions have been described earlier. [8] The PCR product was run on 1.5% agarose gel stained with ethidium bromide. Specifi c bands of 123 bp for IS6110 and 240 bp for MPB64 were visualized using a digital gel documentation system (Alpha Innotech, San Leandro, CA, USA) as shown in Figure 1.…”

“…This method is cost-effective and chances of contamination are also less as shown in our previous experience with osteoarticular and meningeal tuberculosis. [7,8] We have not been able to ascertain if others have used MPCR for TBLA. Therefore, this study was carried out to evaluate MPCR using MPB64 and IS6110, and to compare it with conventional methods such as microscopy, culture, and cytological examination for rapid diagnosis of TBLA.…”

Evaluation of Spondylodiscitis Cases Caused By Tuberculosis

“…Bu yazıda kliniğimize başvuran tüberküloza bağlı spondilodiskit olguları değerlendirilmesi sunulmuştur. (7)(8). Konvensiyonel radyoloji, kas iskelet tüberkülozu olgularının tanısında anahtar role sahiptir.…”

Evaluation of Microscopy, Culture and PCR Methods in the Laboratory Diagnosis of Genito-urinary Tuberculosis

“…However, few studies from different geographical regions of the world have reported that some clinical isolates have either a single copy or no copy of IS6110 which leads to false negative results [24,25]. For avoiding the contamination during amplification and increasing the sensitivity, new approaches such as nested PCR (two step amplification) [26,27] and multiplex PCR (amplification of two or more gene targets simultaneously) [28,29,30,31] have been exploited for EPTB diagnosis.…”