Journal List > Korean J Clin Microbiol > v.14(3) > 1038223

Wang, Jin, Bang, Choi, Park, Koh, and Lee: Evaluation of MolecuTech Real MTB-ID for MTB/NTM Detection Using Direct Specimens

Abstract

Background

The differentiation of Mycobacterium tuberculosis (MTB) from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. The diagnosis of diseases caused by NTM is difficult because NTM are prevalent in the environment and have fastidious properties. In this study, we evaluated the realtime PCR-based MTB/NTM detection kit for its usefulness in discrimination of MTB and NTM species.

Methods

A total of 155 sputum specimens whose AFB staining smear and culture were positive were used for this study. Among them, 59 and 96 samples had been identified as MTB and NTM, respectively. DNA obtained from sputum specimens was subjected to analysis with MolecuTech Real MTB-ID (M&D, Korea) realtime PCR-based MTB/NTM detection kit. Subsequently, the results of MolecuTech Real MTB-ID were compared with AFB staining smear and culture results.

Results

The positive rate of MolecuTech Real MTB-ID to detect MTB and NTM was 98.3% (58/59) and 97.9 (94/96), respectively, using sputum specimens.

Conclusion

For detection of MTB/NTM, the sensitivity and specificity of MolecuTech Real MTB-ID were comparable to those of conventional methods. Therefore, this study suggests the usefulness of realtime PCR-based MolecuTech Real MTB-ID for rapid detection of MTB/NTM from direct specimens.

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Fig. 1.
The sensitivity of the realtime PCR for detecting M. tuberculosis H37Rv (A) and M. intracellulare (B) genomic DNA (A) using FAM dye channel for detecting MTB complex (B) and using ROX dye channel for detecting NTM. Real-time PCR with 1 ng (D3), 100 pg (D4), 10 pg (D5), 1 pg (D6), 100 fg (D7), 10 fg (D8), 1 fg (D9).
kjcm-14-103f1.tif
Table 1.
Smear and MTB/NTM differentiation results of sputum specimens used in this study
AFB smear Culture confirmation results Total number of sputum specimens
MTB NTM
Trace 3 5 8
+1 22 24 46
+2 12 32 44
+3 6 15 21
+4 13 20 36
Total 59 96 155

Abbreviations: AFB, acid-fast bacilli; MTB, M. tuberculosis; NTM, nontuberculous mycobacteria.

Table 2.
Comparison of MolecuTech Real-MTB ID using sputum specimens
AFB smear MolecuTech Real-MTB ID Total
MTB (∗) Ct NTM Ct ND
Trace 2 31.8-35.06 5 28.1-35.2 1 8
1+ 22 (1) 28.8-35.3 23 26.6-35.7 1 46
2+ 12 (1) 25.8-35.5 31 21.7-35.6 1 44
3+ 6 (1) 23.8-30.4 15 27.6-35.02   21
4+ 16 20.4-28.8 20 23.5-34.9   36
Total 58 (98.3%)   94 (97.9%)   3 (1.9%) 155

(∗): number of MTB/NTM mixed case. Abbreviations: Ct, threshold cycle; ND, not determined.

Table 3.
Confirmation of mixed presence of MTB/NTM in sputum by using REBA Myco-ID samples tested in this experiment were the ones determined to contain MTB and NTM by MolecuTech Real-MTB ID
kjcm-14-103t3.tif
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