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Automated liquid culture system misses isoniazid heteroresistance in Mycobacterium tuberculosis isolates with mutations in the promoter region of the inhA gene

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Abstract

Heteroresistance in Mycobacterium tuberculosis isolates remains the major challenge for phenotypic drug susceptibility testing (DST) methods to detect drug resistance. The aim of this study was to investigate the abilities of phenotypic DST methods to identify the isoniazid (INH) heteroresistance in M. tuberculosis. We found that the broth dilution method was able to detect INH resistance if 0.5 % resistant bacteria with mutations in the katG and oxyR-ahpC regions were present, while the detection limit ranged from 1 to 10 % for the INH-resistant strains harboring inhA mutations, which was associated with the different mutant types. Additionally, MGIT DST was able to find the recommended 1 % INH resistance due to katG mutations. In contrast, MGIT DST detected resistance in suspensions with 20 % resistant bacteria with inhA mutations. Statistical analysis revealed that the ability of the broth dilution method to detect heteroresistance was better than that of the MGIT DST (p = 0.004). When we further pairwise compared the two methods for detecting heteroresistance according to different mutant loci, the broth dilution method found more heteroresistance due to inhA mutations than MGIT DST (p = 0.001), while the differences for katG and oxyR-ahpC mutations were both not statistically significant (p > 0.05). In conclusion, our findings demonstrate that MGIT DST fails to detect INH heteroresistance in M. tuberculosis isolates with mutations in the promoter region of inhA. In addition, the broth dilution method is more sensitive than MGIT DST in finding INH heteroresistance, indicating that this method may serve as an alternative method to detect the heteroresistance of M. tuberculosis.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (81301509) and National Key Research Program of China (2013ZX10003-003). We are grateful to the members of the National Tuberculosis Reference Laboratory at the Chinese Center for Disease Control and Prevention for their cooperation and technical help.

Conflict of interest

The authors declare that they have no competing interests.

Author information

Authors and Affiliations

  1. Respiratory Diseases Department of Nanlou, Chinese People’s Liberation Army General Hospital, Beijing, China

    Z. Zhang & Y. Pang

  2. National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Chang Bai Road, Changping District, Beijing, 102206, People’s Republic of China

    Z. Zhang, Y. Wang, Y. Pang & Y. Zhao

  3. Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, Beijing, China

    J. Lu

Authors
  1. Z. Zhang
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  2. J. Lu
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  3. Y. Wang
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  4. Y. Pang
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  5. Y. Zhao
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Corresponding authors

Correspondence to Y. Pang or Y. Zhao.

Additional information

Zhijian Zhang and Jie Lu contributed equally to this paper.

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Zhang, Z., Lu, J., Wang, Y. et al. Automated liquid culture system misses isoniazid heteroresistance in Mycobacterium tuberculosis isolates with mutations in the promoter region of the inhA gene. Eur J Clin Microbiol Infect Dis 34, 555–560 (2015). https://doi.org/10.1007/s10096-014-2262-0

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  • DOI: https://doi.org/10.1007/s10096-014-2262-0

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