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Hahm, So, Chung, and Lee: Evaluation of the AdvanSure TB/NTM Plus Real-time PCR Assay for the Simultaneous Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria from Clinical Specimens
Original Article

Ann Clin Microbiol 2020;23(2):105-116.

Published online: 30 June 2020

DOI: https://doi.org/10.5145/ACM.2020.23.2.7

Evaluation of the AdvanSure TB/NTM Plus Real-time PCR Assay for the Simultaneous Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria from Clinical Specimens

Chorong Hahm1, 2, Min-Kyung So1, Hae-Sun Chung1, Miae Lee1

1Department of Laboratory Medicine, College of Medicine, Ewha Womans University, Seoul,Korea

2Eone Laboratories, Seoul,Korea

Corresponding author: Miae Lee E-mail: miae@ewha.ac.kr

Received 5 March 2020       Revised 28 April 2020       Accepted 14 May 2020

© Korean Society of Clinical Microbiology. All rights reserved.

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/.

Abstract

Background: The AdvanSure TB/NTM plus real-time PCR (AdvanSure plus PCR; LG Chem., Seoul, Korea) assay has been developed to increase the diagnostic sensitivity of nontuberculous mycobacteria (NTM) compared with the currently used The AdvanSure TB/NTM real-time PCR (AdvanSure PCR; LG Chem., Seoul, Korea) assay. In this study, we aimed to evaluate the performance of the AdvanSure plus PCR comparing the results with mycobacterial culture and the AdvanSure. Methods: Patients (n=199) with suspected NTM or Mycobacterium tuberculosis complex (MTC) were tested using AdvanSure plus PCR, AdvanSure, acid-fast bacilli staining, and mycobacteria culture. Additionally, 200 DNA samples (n=100, MTC and n=100, NTM) were obtained from positive MTC or NTM cultures for evaluation using the AdvanSure plus PCR assay. Results: The two real-time PCR systems showed a 94.0% (n=187/199) concordance rate (Kappa=0.94). Based on culture results, the sensitivity and specificity for the detection of v were 100% (45/45) and 83.8% (129/154) using AdvanSure plus PCR, and 100.0% (45/45) and 99.3% (153/154) using AdvanSure, respectively. The sensitivity and specificity for NTM detection were 68.0% (n=17/25) and 90.2% (n=157/174), respectively, with AdvanSure plus PCR, and 64.0% (n=16/25) and 95.4% (n=166/174), respectively, using AdvanSure. With culture-positive samples, AdvanSure plus PCR tested positive for 100% of both the MTC and NTM specimens. Seven (out of 200) culture-positive samples tested positive for both MTC and NTM using the AdvanSure plus PCR. Conclusion: AdvanSure plus PCR had increased sensitivity but decreased specificity compared with AdvanSure for the detection of NTM. The AdvanSure plus PCR assay can be used for the simultaneous detection of MTC and NTM in direct specimen and culture.

Keywords: Mycobacterium, Polymerase chain reaction, Tuberculosis

CONFLICTS OF INTEREST

No potential conflicts of interest relevant to this article were reported.

FUNDING

본 연구는 LG Chem.의 연구비 지원을 받아 수행되었으나 본 연구의 방법 및 결과 등에는 관여하지 않았습니다.

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Table 1
Frequency of Mycobacterium species in this study
acm_2302_006-t1.png

Abbreviation: MTC, M. tuberculosis complex; NTM, nontuberculous mycobacteria.

Table 2
Performance of AdvanSure plus PCR in 200 mycobacterial strains.
acm_2302_006-t2.png

Abbreviations: MTC, M. tuberculosis complex; NTM, nontuberculous mycobacteria; AdvanSure plus PCR, AdvanSure TB/NTM plus real-time PCR

Table 3
Results of AdvanSure plus PCR and AdvanSure PCR in 7 cases of MTC and NTM mix
acm_2302_006-t3.png

Abbreviation: Ct, threshold cycle; IC, internal control; MTC, M. tuberculosis complex; NTM, nontuberculous mycobacteria; AdvanSure PCR, AdvanSure TB/NTM real-time PCR; AdvanSure plus PCR, AdvanSure TB/NTM plus real-time PCR.

Table 4
Results of AdvanSure plus PCR, AdvanSure PCR assay and AFB staining according to the results of culture
acm_2302_006-t4.png

Abbreviation: MTC, M. tuberculosis complex; NTM, nontuberculous mycobacteria; AdvanSure PCR, AdvanSure TB/NTM real-time PCR; AdvanSure plus PCR, AdvanSure TB/NTM plus real-time PCR.

Table 5
The performance of AdvanSure plus PCR, AdvanSure PCR assay, and AFB stain for detection of MTC/NTM compared with mycobacterial culture
acm_2302_006-t5.png

*Included negative culture samples (n=129) and NTM positive culture samples (n=25).†Included MTC & NTM co-positive samples (n=3) by AdvanSure plus PCR.‡Included negative culture samples (n=129) and MTC positive culture samples (n=45).§Calculated using Korean tuberculosis prevalence of 0.0768%.Abbreviation: MTC, M. tuberculosis complex; NTM, nontuberculous mycobacteria; AdvanSure PCR, AdvanSure TB/NTM real-time PCR; AdvanSure plus PCR, AdvanSure plus TB/NTM real-time PCR.

Table 6
Concordance of AdvanSure plus PCR, AdvanSure PCR assay for detection of MTC and NTM
acm_2302_006-t6.png

Abbreviation: MTC, M. tuberculosis complex; NTM, nontuberculous mycobacteria; AdvanSure PCR, AdvanSure TB/NTM real-time PCR; AdvanSure plus PCR, AdvanSure TB/NTM plus real-time PCR.

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