Evaluation of MolecuTech Real MTB-ID for MTB/NTM Detection Using Direct Specimens

Hyeyoung Wang; Hyunwoo Jin; Hyeeun Bang; Yeon-Im Choi; Eun-mi Park; Won-jung Koh; Hyeyoung Lee

doi:10.5145/KJCM.2011.14.3.103

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

Original Article

Korean J Clin Microbiol 2011;14(3):103-109.

Published online: 21 September 2011

DOI: https://doi.org/10.5145/KJCM.2011.14.3.103

Evaluation of MolecuTech Real MTB-ID for MTB/NTM Detection Using Direct Specimens

Hyeyoung Wang^1,^∗, Hyunwoo Jin^2,^∗, Hyeeun Bang², Yeon-Im Choi², Eun-mi Park³, Won-jung Koh⁴, Hyeyoung Lee²

¹M&D, Inc., Wonju, Korea

²Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea

³YD Diagnostics, Yongin, Korea

⁴Division of Pulmonary and Critical Care Medicine, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea

Correspondence: Hyeyoung Lee, Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, 234 Maeji-ri, Heungup-myeon, Wonju 220-710, Korea. (Tel) 82-33-760-2740, (Fax) 82-33-760-2561, (E-mail) hyelee@yonsei.ac.kr

^∗ These authors are equally contributed.

Received 3 March 2011 Revised 23 June 2011 Accepted 22 July 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Mycobacterium tuberculosis, Non-tuberculous mycobacteria, Real-time PCR

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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).

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
AFB smear	MTB	NTM	Total number of sputum specimens
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
AFB smear	MTB (∗)	Ct	NTM	Ct	ND	Total
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

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