Evaluation Of Multiplex Polymerase Chain Reaction Using Two Primers And Its Comparison With Conventional Isolation Techniques For The Diagnosis Of...

1036 words - 5 pages

Tuberculous meningitis is a serious hazard to public health especially in developing countries because of the devastating neurological sequelae and associated high mortality. It is very important to diagnose TBM in early stage to prevent such consequences. But the diagnosis still remains difficult because of the absence of a reliable and rapid technique. Sharma et al.20 reported sensitivity of ZN smear and LJ culture to be 1.4% and 14.3% respectively. The lower sensitivity is due to the paucibacillary nature of the disease as the number of organisms required for smear to be positive is 104 organisms per ml. Therefore, a sufficient amount of CSF must be processed. Previous studies have shown that sensitivity of AFB smear for diagnosis of TBM varies between 5-30%.1, 21, 22 The sensitivity of ZN smear in the present study was also low i.e 3.6% which is similar to the earlier reports. Centrifugation of large volumes (10–20 ml) of CSF for 30 min in a case of TBM and smear examination from the deposit improves the detection rate of AFB.23, 24
We observed higher sensitivity of BACTEC MGIT 960 (16.4%) over conventional LJ culture (5.5%). The 2 cases which were smear positive were negative on LJ culture but positive in BACTEC. The conventional LJ culture of M. tuberculosis from CSF is insensitive and slow (4-6 weeks). The sensitivity of LJ culture reported in earlier studies ranged from 10-83%.21, 25 The liquid BACTEC culture which is an automated system for diagnosis of TBM has a sensitivity ranging from 4.3 to 48.9%.21 Our findings are consistent with these studies. Mean duration of positivity of LJ culture was 28 days and by BACTEC culture, it was 13 days which is consistent with earlier studies.21, 26 Our results emphasized the fact that conventional modalities including BACTEC have not only poor sensitivity but they also take longer time to give positive results making these tests less useful to the clinician in the diagnosis of TBM.
Nucleic acid amplification tests (NAATs) have been found to be most sensitive and specific method among the available diagnostic techniques. In the literature, wide variability is seen with regard to the sensitivity of PCR. The reported sensitivity of the PCR technique in CSF varies from 33% to 90% and the specificity from 88% to 100%27 (Table 7).
Table I: Sensitivity and specificity of PCR in CSF of TBM cases in previous studies
Study, year Primer Sensitivity Specificity
Shankar et al. 199128
MPB64 65% 88%
Lee et al. 199429
IS6110 100% 38%

MPB64 100% 90%

65kDa 100% 62%

Seth et al. 199630
MPB64 88% 94%

Smith et al. 199631
IS6110 25% 100%

Narayanan et al. 200132
IS6110 95% 75%

TRC4 100% 75%

Sumi et al. 200233
IS6110 47% 100%
Kulkarani et al. 20054
Pab 90% 100%
Takahashi et al. 200534
Nested MPB64 100% 100%
Deshpande et al. 200735
IS6110 82.4% 75.9%
Rafi et al. 200736
IS6110 76.3% 89.18%
Afroze et al. 200837
MPB64 77.7% 100%
Sharma et al. 201020
Pab 82.8% 100%
Sharma et al. 201238
MPB64 82.22% 100%


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