Volume 10,Issue 1
Fall 2025
Evaluation of the AdvanSure TB/NTM Plus Real-Time PCR Assay for the Simultaneous Detection of Mycobacterium tuberculosis and Nontuberculous Mycobacteria from Clinical Specimens
Background: The AdvanSure TB/NTM plus real-time PCR (AdvanSure plus PCR; LG Chem., Korea) assay has been developed to increase the diagnostic sensitivity of nontuberculous mycobacteria (NTM) compared with the existing AdvanSure TB/NTM real-time PCR (AdvanSure PCR; LG Chem., Korea) assay. This study aimed to evaluate the performance of the AdvanSure plus PCR comparing the results with mycobacterial culture and the AdvanSure PCR. Methods: Patients (n = 199) with suspected NTM or Mycobacterium tuberculosis complex (MTC) were tested using AdvanSure plus PCR assay,AdvanSure PCR assay, acid-fast bacilli staining, and mycobacteria culture. Additionally,200 DNA samples (MTC, n = 100, and NTM, n = 100) 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 MTC were 100% (n = 45/45) and 83.8% (n = 129/154) using AdvanSure plus PCR, and 100.0% (n = 45/45), and 99.3% (n = 153/154) using AdvanSure PCR, respectively. The sensitivity and specificity for NTM detection were 68.0% (n = 17/25) and 90.2% (n = 157/174) using AdvanSure plus PCR, and 64.0% (n = 16/25) and 95.4% (n = 166/174) using AdvanSure PCR, respectively. 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 PCR for the detection of NTM. The AdvanSure plus PCR assay can be used for the simultaneous detection of MTC and NTM in direct specimens and cultures.
1. 2000, Diagnostic Standards and Classification of Tuberculosis in Adults and Children. Am J Respir Crit Care Med, 161: 1376–1395.
2. Lee H, Myung W, Koh WJ, et al., 2019, Epidemiology of Nontuberculous Mycobacterial Infection, South Korea, 2007-2016. Emerg Infect Dis, 25: 569–572.
3. Dye C, Watt CJ, Bleed DM, et al., 2005, Evolution of Tuberculosis Control and Prospects for Reducing Tuberculosis Incidence, Prevalence, and Deaths Globally. JAMA, 293: 2767–2775.
4. World Health Organization. Global tuberculosis report 2019. World Health Organization, 2019. Viewed 17 Oct 2019, http://www.who.int/tb/publications/global_report/en/
5. Levy H, Feldman C, Sacho H, et al., 1989, A Reevaluation of Sputum Microscopy and Culture in the Diagnosis of Pulmonary Tuberculosis. Chest, 95: 1193–1197.
6. Ritchie SR, Harrison AC, Vaughan RH, et al., 2007, New Recommendations for Duration of Respiratory Isolation Based on Time to Detect Mycobacterium tuberculosis in Liquid Culture. Eur Respir J, 30: 501–507.
7. Greco S, Rulli M, Girardi E, et al., 2009, Diagnostic Accuracy of In-House PCR for Pulmonary Tuberculosis in Smear-Positive Patients: Meta-Analysis and Metaregression. J Clin Microbiol, 47: 569–576.
8. Bartlett JM, Stirling D, 2003, A Short History of the Polymerase Chain Reaction. Methods Mol Biol, 226: 3–6.
9. Kim SW, Kim SI, Lee SJ, et al., 2015, The Effectiveness of Real-Time PCR Assay, Compared with Microbiologic Results for the Diagnosis of Pulmonary Tuberculosis. Tuberc Respir Dis, 78: 1–7.
10. Drosten C, Panning M, Kramme S, 2003, Detection of Mycobacterium tuberculosis by Real-Time PCR Using Pan-Mycobacterial Primers and a Pair of Fluorescence Resonance Energy Transfer Probes Specific for the M. tuberculosis Complex. Clin Chem, 49: 1659–1661.
11. Jung CL, Kim MK, Seo DC, et al., 2008, Clinical Usefulness of Real-Time PCR and Amplicor MTC PCR Assays for Diagnosis of Tuberculosis. Korean J Clin Microbiol, 11: 29–33.
12. Ho TBL, Robertson BD, Taylor GM, et al., 2000, Comparison of Mycobacterium tuberculosis Genomes Reveals Frequent Deletions in a 20 kb Variable Region in Clinical Isolates. Int J Genomics, 1: 272–282.
13. Talarico S, Durmaz R, Yang Z. 2005, Insertion- and Deletion-Associated Genetic Diversity of Mycobacterium tuberculosis Phospholipase C-Encoding Genes Among 106 Clinical Isolates from Turkey. J Clin Microbiol, 43: 533–538.
14. Gordon SV, Brosch R, Billault A, et al., 1999, Identification of Variable Regions in the Genomes of Tubercle Bacilli using Bacterial Artificial Chromosome Arrays. Mol Microbiol, 32: 643–655.
15. Wang HY, Jin HW, Bang HE, et al., 2011, Evaluation of MolecuTechreal MTC-ID for MTC/NTM Detection using Direct Specimens. Korean J Clin Microbiol, 14: 103–109.
16. Mäkinen J, Marjamäki M, Marttila H, et al., 2006, Evaluation of a Novel Strip Test, GenoType Mycobacterium CM/AS, for Species Identification of Mycobacterial Cultures. Clin Microbiol Infect, 12: 481–483.
17. Hwang S, Oh KJ, Jang IH, et al., 2011, Evaluation of the Diagnostic Performance of the AdvanSure TB/NTM Real-Time PCR Kit for Detection of Mycobacteria. Korean J Clin Microbiol, 14: 55–59.
18. Mdivani N, Li H, Akhalaia M, et al., 2009, Monitoring Therapeutic Efficacy by Real-Time Detection of Mycobacterium tuberculosis mRNA in Sputum. Clin Chem, 55: 1694–1700.
19. Rosso F, Michelon CT, Sperhacke RD, et al., 2011, Evaluation of Real-Time PCR of Patient Pleural Effusion for Diagnosis of Tuberculosis. BMC Res Notes, 4: 279.
20. Thomsen VO, Kok-Jensen A, Buser M, et al., 1999, Monitoring Treatment of Patients with Pulmonary Tuberculosis: Can PCR be Applied? J Clin Microbiol, 37: 3601–3607.
21. Forbes BA, Hall GS, Miller MB, et al., 2018, Practice Guidelines for Clinical Microbiology Laboratories: Mycobacteria. Clin Microbiol Rev, 31: e00038-17.
22. Min JW, Yoon HI, Park KU, et al., 2010, Real-Time Polymerase Chain Reaction in Bronchial Aspirate for Rapid Detection of Sputum Smear-Negative Tuberculosis. Int J Tuberc Lung Dis, 14: 852–858.