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Rapid detection of fluoroquinolone resistance in Mycobacterium tuberculosis using a novel multienzyme isothermal rapid assay


Simple, rapid, and accurate detection of Fluoroquinolone (FQ) resistance is essential for early initiation of appropriate anti-tuberculosis treatment regimen among rifampicin-resistant tuberculosis (RR-TB). In this study, we developed a new assay, which combines multienzyme isothermal rapid amplification and a lateral flow strip (MIRA-LF), to identify the mutations on codons 90 and 94 of gyrA for detecting levofloxacin (LFX) resistance. Compared to conventional phenotypic drug susceptibility testing, the new assay detected fluoroquinolone resistance with a sensitivity, specificity, and accuracy of 92.4%, 98.5%, and 96.5%, respectively. Thus, these characteristics of the newly developed MIRA-LF assay make it particularly useful and accurate for detecting FQ resistance in Mycobacterium tuberculosis in resource-limited condition.

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Fig. 1


  1. Global tuberculosis report 2022. Geneva: World Health Organization; 2022. licence: cc bY-Nc-sa 3.0 iGo.

  2. WHO operational handbook on tuberculosis. Module 4: treatment - drug-resistant tuberculosis treatment. Geneva: World Health Organization; 2020. Licence: CC BY-NC-SA 3.0 IGO.

  3. Chien JY, Chiu WY, Chien ST, Chiang CJ, Yu CJ, Hsueh PR. Mutations in gyrA and gyrB among Fluoroquinolone- and Multidrug-Resistant Mycobacterium tuberculosis Isolates. Antimicrob Agents Chemother. 2016;60:2090–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Willby M, Sikes RD, Malik S, Metchock B, Posey JE. Correlation between GyrA substitutions and ofloxacin, levofloxacin, and moxifloxacin cross-resistance in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2015;59:5427–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Avalos E, Catanzaro D, Catanzaro A, Ganiats T, Brodine S, Alcaraz J, Rodwell T. Frequency and geographic distribution of gyrA and gyrB mutations associated with fluoroquinolone resistance in clinical Mycobacterium tuberculosis isolates: a systematic review. PLoS One. 2015;10:e0120470.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Heng P, Liu J, Song Z, Wu C, Yu X, He Y. Rapid detection of Staphylococcus aureus using a novel multienzyme isothermal rapid amplification technique. Front Microbiol. 2022;13:1027785.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Lu Y, Li MC, Liu HC, Lin SQ, Zhao XQ, Liu ZG, Zhao LL, Wan KL. Detecting Mycobacterium tuberculosis complex and rifampicin resistance via a new rapid multienzyme isothermal point mutation assay. Anal Biochem. 2021;630:114341.

    Article  CAS  PubMed  Google Scholar 

  8. Technical Report on critical concentrations for drug susceptibility testing of medicines used in the treatment of drug-resistant tuberculosis. Geneva: World Health Organization; 2018 (WHO/CDS/TB/2018.5). Licence: CC BY-NC-SA 3.0 IGO.

  9. Zhao LL, Chen Y, Liu HC, Xia Q, Wu XC, Sun Q, Zhao XQ, Li GL, Liu ZG, Wan KL. Molecular characterization of multidrug-resistant Mycobacterium tuberculosis isolates from China. Antimicrob Agents Chemother. 2014;58:1997–2005.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Rufai SB, Singh J, Kumar P, Mathur P, Singh S. Association of gyrA and rrs gene mutations detected by MTBDRsl V1 on Mycobacterium tuberculosis strains of diverse genetic background from India. Sci Rep. 2018;8:9295.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Javed H, Bakula Z, Plen M, Hashmi HJ, Tahir Z, Jamil N, Jagielski T. Evaluation of genotype MTBDRplus and MTBDRsl assays for rapid detection of drug resistance in extensively drug-resistant Mycobacterium tuberculosis isolates in Pakistan. Front Microbiol. 2018;9:2265.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Penn-Nicholson A, Georghiou SB, Ciobanu N, Kazi M, Bhalla M, David A, Conradie F, Ruhwald M, Crudu V, Rodrigues C, Myneedu VP, Scott L, Denkinger CM, Schumacher SG, Xpert XDRTC. Detection of isoniazid, fluoroquinolone, ethionamide, amikacin, kanamycin, and capreomycin resistance by the Xpert MTB/XDR assay: a cross-sectional multicentre diagnostic accuracy study. Lancet Infect Dis. 2022;22:242–9.

    Article  CAS  PubMed  Google Scholar 

  13. Cao Y, Parmar H, Gaur RL, Lieu D, Raghunath S, Via N, Battaglia S, Cirillo DM, Denkinger C, Georghiou S, Kwiatkowski R, Persing D, Alland D, Chakravorty S. Xpert MTB/XDR: a 10-color reflex assay suitable for point-of-care settings to detect isoniazid, fluoroquinolone, and second-line-injectable-drug resistance directly from Mycobacterium tuberculosis-positive sputum. J Clin Microbiol. 2021;59:e02314–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Bablishvili N, Tukvadze N, Shashkina E, Mathema B, Gandhi NR, Blumberg HM, Kempker RR. Impact of gyrB and eis mutations in improving detection of second-line-drug resistance among Mycobacterium tuberculosis isolates from Georgia. Antimicrob Agents Chemother. 2017;61:e01921–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Goossens SN, Sampson SL, Van Rie A. Mechanisms of drug-induced tolerance in Mycobacterium tuberculosis. Clin Microbiol Rev. 2020;34:e00141–20.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Huo F, Ma Y, Li S, Xue Y, Shang Y, Dong L, Li Y, Pang Y. Specific gyrA gene mutations correlate with high prevalence of discordant levofloxacin resistance in Mycobacterium tuberculosis isolates from Beijing, China. J Mol Diagn. 2020;22:1199–204.

    Article  CAS  PubMed  Google Scholar 

  17. Niward K, Angeby K, Chryssanthou E, Paues J, Bruchfeld J, Jureen P, Giske CG, Kahlmeter G, Schon T. Susceptibility testing breakpoints for Mycobacterium tuberculosis categorize isolates with resistance mutations in gyrA as susceptible to fluoroquinolones: implications for MDR-TB treatment and the definition of XDR-TB. J Antimicrob Chemother. 2016;71:333–8.

    Article  CAS  PubMed  Google Scholar 

  18. Zhang Z, Lu J, Wang Y, Pang Y, Zhao Y. Prevalence and molecular characterization of fluoroquinolone-resistant Mycobacterium tuberculosis isolates in China. Antimicrob Agents Chemother. 2014;58:364–9.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Rigouts L, Coeck N, Gumusboga M, de Rijk WB, Aung KJ, Hossain MA, Fissette K, Rieder HL, Meehan CJ, de Jong BC, Van Deun A. Specific gyrA gene mutations predict poor treatment outcome in MDR-TB. J Antimicrob Chemother. 2016;71:314–23.

    Article  CAS  PubMed  Google Scholar 

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This study was supported by the projects from National Key Program of Mega Infectious Diseases (Grant No. 2018ZX10302302-001). The funder had no role in the study design, data collection and analysis, decision to publish, or manuscript preparation.

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Correspondence to Li-li Zhao.

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The authors declare no competing interests.

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This study was approved by the Ethics Committee of the National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention. The patients with TB were included only after we received informed written consent from themselves.

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Li, Mc., Lu, Y., Liu, Hc. et al. Rapid detection of fluoroquinolone resistance in Mycobacterium tuberculosis using a novel multienzyme isothermal rapid assay. J Antibiot 76, 598–602 (2023).

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