Ann Clin Microbiol.  2014 Dec;17(4):115-122. 10.5145/ACM.2014.17.4.115.

Performance Evaluation of Anyplex Plus MTB/NTM and MDR-TB Detection Kit for Detection of Mycobacteria and for Anti-Tuberculosis Drug Susceptibility Test

• Lee JH
• Kim BH
• Lee MK

• KMID: 1971106
• DOI: http://doi.org/10.5145/ACM.2014.17.4.115

Abstract

BACKGROUND
The Anyplex plus MTB/NTM and MDR-TB Detection kits (Seegene, Korea) a real-time PCR assays for direct detection of Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) and for identification of rifampin (RIF) and isoniazid (INH) resistance of MTB in various specimens. We evaluated the diagnostic performance of the Anyplex plus MTB/NTM and MDR-TB Detection kit.
METHODS
To determine the ability of the kit to detect MTB and NTM, 557 samples were tested. The diagnostic performance of the Anyplex plus MTB/NTM Detection kit was determined based on the results of culture, nucleic acid amplification tests (NAAT), radiologic analysis, and clinical features suggestive of mycobacterial infection. The performance of the kit was compared with those of other real-time PCR kits. For the drug susceptibility test (DST), 51 MTB isolates were tested. The diagnostic performance of the Anyplex plus MDR-TB Detection kit was determined based on the conventional DST and compared with other molecular DST kits.
RESULTS
Sensitivity and specificity for MTB detection of the Anyplex plus MTB/NTM Detection kit were 82.9% (63/76) and 99.4% (478/481), respectively, while those for NTM detection were 76.5% (13/17) and 89.6% (484/540). Sensitivity and specificity for RIF resistance detection of the Anyplex plus MDR-TB Detection kit were 100% (3/3) and 97.9% (47/48), respectively, while those for INH resistance detection were 83.3% (5/6) and 100% (45/45).
CONCLUSION
The Anyplex plus MTB/NTM Detection kit showed good diagnostic performance for detection of MTB and NTM. Especially in MTB-positive cases, the Anyplex plus MDR-TB Detection kit provided rapid and reliable results of drug resistance to RIF and INH.

Keyword

Anyplex plus; Molecular diagnostic techniques; Multidrug-resistant tuberculosis; Mycobacterium tuberculosis; Nontuberculous Mycobacteria

MeSH Terms

Drug Resistance
Isoniazid
Molecular Diagnostic Techniques
Mycobacterium tuberculosis
Nontuberculous Mycobacteria
Nucleic Acid Amplification Techniques
Real-Time Polymerase Chain Reaction
Rifampin
Tuberculosis, Multidrug-Resistant
Isoniazid
Rifampin

Reference

1.The Korean Society of Clinical Microbiology and Korea Centers for Disease Control and Prevention. Manual of Laboratory Tests for Tuberculosis (2013). http://www.cdc.go.kr/CDC/cms/cmsFile-Download.jsp?fid=51&cid=21373&fieldName=attach1&index=1. [Online] (last visited on 16 December 2014).

2.The Korean Society of Clinical Microbiology and Korea Centers for Disease Control and Prevention. Practical Guidelines for Laboratory Diagnosis of Tuberculosis (2011). https://kslm.org/board/lib/down.php?boardid=board_notice&no=722. [Online] (last visited on 16 December 2014).

3.Diagnostic Standards and Classification of Tuberculosis in Adults and Children. Am J Respir Crit Care Med. 2000. 161:1376–95.

4.Levidiotou S., Vrioni G., Galanakis E., Gesouli E., Pappa C., Stefanou D. Four-year experience of use of the Cobas Amplicor system for rapid detection of Mycobacterium tuberculosis complex in respiratory and nonrespiratory specimens in Greece. Eur J Clin Microbiol Infect Dis. 2003. 22:349–56.

5.Report of an expert consultation on the use of nucleic acid amplification tests for the diagnosis of tuberculosis: Centers for Disease Control and Prevention. http://www.cdc.gov/tb/publications/guidelines/am-plification_tests/default.htm. [Online] (last visited on 1 September 2012).

6.Lim J., Kim J., Kim JW., Ihm C., Sohn YH., Cho HJ, et al. Multicenter evaluation of Seegene Anyplex TB PCR for the detection of Mycobacterium tuberculosis in respiratory specimens. J Microbiol Biotechnol. 2014. 24:1004–7.

7.Steingart KR., Henry M., Ng V., Hopewell PC., Ramsay A., Cunningham J, et al. Fluorescence versus conventional sputum smear microscopy for tuberculosis: a systematic review. Lancet Infect Dis. 2006. 6:570–81.

8.Hwang S., Oh KJ., Jang IH., Uh Y., Yoon KJ., Kim HY, et al. Evaluation of the diagnostic performance of the advansure TB/NTM real-time PCR kit for detection of mycobacteria. Korean J Clin Microbiol. 2011. 14:55–9.

9.Telenti A., Imboden P., Marchesi F., Lowrie D., Cole S., Colston MJ, et al. Detection of rifampicin-resistance mutations in Mycobacterium tuberculosis. Lancet. 1993. 341:647–50.

10.Telenti A. Genetics and pulmonary medicine. 5. Genetics of drug resistant tuberculosis. Thorax. 1998. 53:793–7.

11.Heym B., Alzari PM., Honoré N., Cole ST. Missense mutations in the catalase-peroxidase gene, katG, are associated with isoniazid resistance in Mycobacterium tuberculosis. Mol Microbiol. 1995. 15:235–45.

12.Zhang Y., Heym B., Allen B., Young D., Cole S. The catalase-per-oxidase gene and isoniazid resistance of Mycobacterium tuberculosis Nature. 1992. 358:591–3.

13.Banerjee A., Dubnau E., Quemard A., Balasubramanian V., Um KS., Wilson T, et al. inhA, a gene encoding a target for isoniazid and ethionamide in Mycobacterium tuberculosis. Science. 1994. 263:227–30.

14.Rozwarski DA., Grant GA., Barton DH., Jacobs WR Jr., Sacchettini JC. Modification of the NADH of the isoniazid target (InhA) from Mycobacterium tuberculosis. Science. 1998. 279:98–102.

15.Ma X., Wang H., Deng Y., Liu Z., Xu Y., Pan X, et al. rpoB Gene mutations and molecular characterization of rifampin-resistant Mycobacterium tuberculosis isolates from Shandong Province, China. J Clin Microbiol. 2006. 44:3409–12.

16.Ohno H., Koga H., Kohno S., Tashiro T., Hara K. Relationship between rifampin MICs for and rpoB mutations of Mycobacterium tuberculosis strains isolated in Japan. Antimicrob Agents Chemother. 1996. 40:1053–6.

17.Taniguchi H., Aramaki H., Nikaido Y., Mizuguchi Y., Nakamura M., Koga T, et al. Rifampicin resistance and mutation of the rpoB gene in Mycobacterium tuberculosis. FEMS Microbiol Lett. 1996. 144:103–8.

18.Yue J., Shi W., Xie J., Li Y., Zeng E., Wang H. Mutations in the rpoB gene of multidrug-resistant Mycobacterium tuberculosis isolates from China. J Clin Microbiol. 2003. 41:2209–12.