Sang Bong Han; Yongjun Jo; Jin Kyung Yu; Yonggoo Kim; Yeon-Joon Park

doi:10.3343/lmo.2012.2.1.6

Journal List > Lab Med Online > v.2(1) > 1057180

Go to TopGo to Top Go to BottomGo to Bottom

TOOLS

Han, Jo, Yu, Kim, and Park: Performance Assessment of Advansure™ MDR-TB Genoblot Assay Kit for Anti-tuberculosis Drug Susceptibility Test

Original Article

Clinical Microbiology

Laboratory Medicine Online

Laboratory Medicine Online 2012; 2(1): 34-40.

Published online: 1 January 2012

DOI: https://doi.org/10.3343/lmo.2012.2.1.6

Performance Assessment of Advansure™ MDR-TB Genoblot Assay Kit for Anti-tuberculosis Drug Susceptibility Test

Sang Bong Han, M.D., Yongjun Jo, M.D., Jin Kyung Yu, M.S., Yonggoo Kim, M.D., Yeon-Joon Park, M.D.

Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea.

Corresponding author: Yeon-Joon Park, M.D. Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul 137-701, Korea. Tel: +82-2-2258-1640, Fax: +82-2-2258-1719, yjpk@catholic.ac.kr

Received 16 July 2011 Revised 9 August 2011 Accepted 12 August 2011

(open-access):

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

Because of the long time required for conventional drug susceptibility test (DST) for rifampin and isoniazid, development of rapid DSTs is necessary. Recently, the AdvanSure™ MDR-TB GenoBlot Assay kit (LG Life Science, Korea), using reverse hybridization line blot assay, was developed. We compared this kit with Genotype® MTBDRplus (HAIN Lifescience, Germany) and conventional DST.

Methods

Of the DNAs preserved after performing DST by using Genotype®, we selected 144 samples having conventional DST results. The experiments with both the kits were performed according to the manufacturers' instructions. For the samples for which discrepant results were obtained, sequencing was performed if the DNA was available. Conventional DST was performed at the Korean Institute of Tuberculosis by using the absolute concentration method.

Results

For rifampin, the findings obtained using both the kits were the same with concordance rates of 98.6% (142/144) compared to conventional DST. Of the 2 discrepant findings, one was very major error and the other was major error. For isoniazid, compared to conventional DST, concordance rates of AdvanSure™ and Genotype® were 95.8%(138/144) and 95.1%(137/144) respectively. Of the 6 discrepant findings between conventional method and Advansure™, 5 were very major error and one was major error. All the 7 discrepant findings between conventional method and Genotype® were very major error.

Conclusions

The findings obtained using AdvanSure™ showed high concordance with those obtained using Genotype® and conventional DST. This kit has a higher rate of detection of isoniazid resistance because it includes probes for an additional target (ahpC).

Keywords: Multidrug-resistant Mycobacterium tuberculosis, Rifampin, Isoniazid, Molecular method

Figures and Tables

Table 1

Mutations in the rpoB, katG, inhA, and ahpC genes and the corresponding wild-type and mutation probes in the AdvanSure™ MDR-TB GenoBlot Assay kit

^*to detect INH high level resistance; ^†to detect INH low level resistance.

Abbreviations: RW, rpoB wild type; RM, rpoB mutation type; KW, katG wild type; KM, katG mutation type; IW, inhA wild type; IM, inhA mutation type; AW, ahpC wild type; AM, ahpC mutation type; INH, isoniazid.

Table 2

Mutations in the rpoB, katG, and inhA genes and the corresponding wild-type and mutation probes in the Genotype® MTBDRplus kit

^*Thus far, this rare mutation has only been detected in silico. Therefore, its detection in vitro may not be possible.

Table 3

The results of drug susceptibility test by the 3 methods

Abbreviations: DST, drug susceptibility test; RIF, rifampin; INH, isoniazid; R, resistant; S, susceptible; AdvanSure™, AdvanSure™ MDR-TB GenoBlot (LG Life Science, Korea); Genotype® MTBDRplus (HAIN Lifescience, Germany).

Table 4

The results of 5 specimens that should be retested

^*Samples could not be retested because of the lack of preserved DNA.

Table 5

The number of mutations associated with RIF and INH resistance detected by AdvanSure™ MDR-TB GenoBlot Assay

Abbreviations: DST, drug susceptibility test; RIF, rifampin; INH, isoniazid; R, resistant; S, susceptible; RW2, rpoB wild-type codon 514-520.

Table 6

Discrepant findings of the three methods

^*Samples could not be retested because of the lack of preserved DNA.

Abbreviations: DST, drug susceptibility test; RIF, rifampin; INH, isoniazid; R, resistant; S, susceptible; r, low resistance; WT, wild type; AM1, ahpC G-6A; AM2, ahpC C-10A; RW2, rpoB wild type.

Notes

This article is available from http://www.labmedonline.org

References

1. Centers for Disease Control and Prevention (CDC). Emergence of Mycobacterium tuberculosis with extensive resistance to second-line drugs -- worldwide, 2000-2004. MMWR Morb Mortal Wkly Rep. 2006. 55:301–305.

2. Bai GH, Park YK, Choi YW, Bai JI, Kim HJ, Chang CL, et al. Trend of anti-tuberculosis drug resistance in Korea, 1994-2004. Int J Tuberc Lung Dis. 2007. 11:571–576.

3. Kim BJ, Lee IH, Lee DH, Bai GH, Kong SJ, Lee SH, et al. The current status of multidrug-resistant tuberculosis in Korea. Tuberc Respir Dis. 2006. 60:404–411.

4. Rossau R, Traore H, De Beenhouwer H, Mijs W, Jannes G, De Rijk P, et al. Evaluation of the INNO-LiPA Rif. TB assay, a reverse hybridization assay for the simultaneous detection of Mycobacterium tuberculosis complex and its resistance to rifampin. Antimicrob Agents Chemother. 1997. 41:2093–2098.

5. Shin DH. New diagnostic methods for Mycobacterium tuberculosis infection. J Korean Med Assoc. 2006. 49:773–780.

6. Murray PR, Baron EJ, editors. Manual of clinical microbiology. 2007. 9th ed. Washington DC: American Society for Microbiology;1255.

7. Huyen MN, Tiemersma EW, Lan NT, Cobelens FG, Dung NH, Sy DN, et al. Validation of the GenoType MTBDRplus assay for diagnosis of multidrug resistant tuberculosis in South Vietnam. BMC Infect Dis. 2010. 10:149.

8. Huang WL, Chen HY, Kuo YM, Jou R. Performance assessment of the GenoType MTBDRplus test and DNA sequencing in detection of multidrug-resistant Mycobacterium tuberculosis. J Clin Microbiol. 2009. 47:2520–2524.

9. Guo Y, Zhou Y, Wang C, Zhu L, Wang S, Li Q, et al. Rapid, accurate determination of multidrug resistance in M. tuberculosis isolates and sputum using a biochip system. Int J Tuberc Lung Dis. 2009. 13:914–920.

10. Sandgren A, Strong M, Muthukrishnan P, Weiner BK, Church GM, Murray MB. Tuberculosis drug resistance mutation database. PloS Med. 2009. 6:e2.

11. Tuberculosis drug resistance mutation database. Updated on April 2010. http://www.tbdreamdb.com/INH.html.

12. Doustdar F, Khosravi AD, Farnia P, Masjedi MR, Velayati AA. Molecular analysis of isoniazid resistance in different genotypes of Mycobacterium tuberculosis isolates from Iran. Microb Drug Resist. 2008. 14:273–279.

13. Hofmann-Thiel S, van Ingen J, Feldmann K, Turaev L, Uzakova GT, Murmusaeva G, et al. Mechanisms of heteroresistance to isoniazid and rifampin of Mycobacterium tuberculosis in Tashkent, Uzbekistan. Eur Respir J. 2009. 33:368–374.

14. Rinder H, Mieskes KT, Löscher T. Heteroresistance in Mycobacterium tuberculosis. Int J Tuberc Lung Dis. 2001. 5:339–345.

15. Woodford N, Ellington MJ. The emergence of antibiotic resistance by mutation. Clin Microbiol Infect. 2007. 13:5–18.

16. Cho EH, Bae HK, Kang SK, Lee EH. Detection of isoniazid and rifampicin resistance by sequencing of katG, inhA, and rpoB genes in Korea. Korean J Lab Med. 2009. 29:455–460.

TOOLS

Similar articles