Genotypic and phenotypic characterization of Mycobacterium tuberculosis resistance against fluoroquinolones in the northeast of Iran

BMC Infectious Diseases - Tập 20 Số 1 - 2020
Mahdieh Sayadi1, Hosna Zare1, Saeid Amel Jamehdar1, Seyed Isaac Hashemy2, Zahra Meshkat1, Saman Soleimanpour1, Sven Hoffner3, Kiarash Ghazvini4
1Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
2Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
3Department of Global Public Health, Karolinska Institute, Stockholm, Sweden
4Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Tóm tắt

Abstract Background Fluoroquinolones are broad-spectrum antibiotics that are recommended, and increasingly important, for the treatment of multidrug-resistant tuberculosis (MDR-TB). Resistance to fluoroquinolones is caused by mutations in the Quinolone Resistance Determining Region (QRDR) of gyrA and gyrB genes of Mycobacterium tuberculosis. In this study, we characterized the phenotypic and genotypic resistance to fluoroquinolones for the first time in northeast Iran. Methods A total of 123 Mycobacterium tuberculosis isolates, including 111 clinical and 12 collected multidrug-resistant isolates were studied. Also, 19 WHO quality control strains were included in the study. The phenotypic susceptibility was determined by the proportion method on Löwenstein-Jensen medium. The molecular cause of resistance to the fluoroquinolone drugs ofloxacin and levofloxacin was investigated by sequencing of the QRDR region of the gyrA and gyrB genes. Results Among 123 isolates, six (4.8%) were fluoroquinolone-resistant according to phenotypic methods, and genotypically three of them had a mutation at codon 94 of the gyrA gene (Asp→ Gly) which was earlier reported to cause resistance. All three remaining phenotypically resistant isolates had a nucleotide change in codon 95. No mutations were found in the gyrB gene. Five of the 19 WHO quality control strains, were phenotypically fluoroquinolone-resistant, four of them were genotypically resistant with mutations at codon 90, 91 of the gyrA gene and one resistant strain had no detected mutation. Conclusions Mutation at codon 94 of the gyrA gene, was the main cause of fluoroquinolone resistance among M. tuberculosis isolates in our region. In 3/6 fluoroquinolone-resistant isolates, no mutations were found in either gyrA or gyrB. Therefore, it can be concluded that various other factors may lead to fluoroquinolone resistance, such as active efflux pumps, decreased cell wall permeability, and drug inactivation.

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Tài liệu tham khảo

WHO. Global tuberculosis report. Geneva: World Health Organization; 2016. Report No.: 924156539X.

Neshani A, Kakhki RK, Sankian M, Zare H, Chichaklu AH, Sayyadi M, et al. Modified genome comparison method: a new approach for identification of specific targets in molecular diagnostic tests using Mycobacterium tuberculosis complex as an example. BMC Infect Dis. 2018;18(1):517.

Kakhki RK, Neshani A, Sankian M, Ghazvini K, Hooshyar A, Sayadi M. The short-chain dehydrogenases/reductases (SDR) gene: a new specific target for rapid detection of Mycobacterium tuberculosis complex by modified comparative genomic analysis. Infect Genet Evol. 2019;70:158–64.

Kaniga K, Cirillo DM, Hoffner S, Ismail NA, Kaur D, Lounis N, et al. A multi-laboratory, multi-country study to determine bedaquiline minimal inhibitory concentration quality control ranges for phenotypic drug-susceptibility testing. J Clin Microbiol. 2016;JCM:01123–16.

Mehrabadi SM, Taraghian M, Pirouzi A, Khaledi A, Neshani A, Rashki S. Pulmonary Nocardiosis in suspected tuberculosis patients: a systematic review and meta-analysis of cross-sectional studies. Ethiop J Health Sci. 2020;30(2):293–300.

Shi W, Forsman LD, Hu Y, Zheng X, Gao Y, Li X, et al. Improved treatment outcome of multidrug-resistant tuberculosis with the use of a rapid molecular test to detect drug resistance in China. Int J Infect Dis. 2020;S1201-9712(20):30265–4.

Dalton T, Cegielski P, Akksilp S, Asencios L, Caoili JC, Cho S-N, et al. Prevalence of and risk factors for resistance to second-line drugs in people with multidrug-resistant tuberculosis in eight countries: a prospective cohort study. Lancet. 2012;380(9851):1406–17.

WHO. WHO Global Task Force outlines measures to combat XDR-TB worldwide. Geneva: WHO; 2006.

Ruan Q, Liu Q, Sun F, Shao L, Jin J, Yu S, et al. Moxifloxacin and gatifloxacin for initial therapy of tuberculosis: a meta-analysis of randomized clinical trials. Emerg Microbes Infect. 2016;5(2):e12.

Izadi N, Samiei A, Ghazvini K. Co-infection of long-standing extensively drug-resistant Mycobacterium tuberculosis (XDR-TB) and non-tuberculosis mycobacteria: a case report. Respir Med Case Rep. 2015;15:12–3.

Zignol M, Dean AS, Alikhanova N, Andres S, Cabibbe AM, Cirillo DM, et al. Population-based resistance of Mycobacterium tuberculosis isolates to pyrazinamide and fluoroquinolones: results from a multicountry surveillance project. Lancet Infect Dis. 2016;16(10):1185–92.

Tagliani E, Cabibbe AM, Miotto P, Borroni E, Toro JC, Mansjö M, et al. Diagnostic performance of the new version (v2. 0) of GenoType MTBDRsl assay for detection of resistance to fluoroquinolones and second-line injectable drugs: a multicenter study. J Clin Microbiol. 2015;53(9):2961–9.

Principi N, Esposito S. Appropriate use of fluoroquinolones in children. Int J Antimicrob Agents. 2015;45(4):341–6.

Bacci C, Galli L, de Martino M, Chiappini E. Fluoroquinolones in children: update of the literature. J Chemother. 2015;27(5):257–65.

Kamal S, Hossain A, Sultana S, Begum V, Haque N, Ahmed J, et al. Anti-tuberculosis drug resistance in Bangladesh: reflections from the first nationwide survey. Int J Tuberc Lung Dis. 2015;19(2):151–6.

Aldred KJ, Blower TR, Kerns RJ, Berger JM, Osheroff N. Fluoroquinolone interactions with Mycobacterium tuberculosis gyrase: enhancing drug activity against wild-type and resistant gyrase. Proc Natl Acad Sci. 2016;113(7):E839–E46.

Fox GJ, Oxlade O, Menzies D. Fluoroquinolone therapy for the prevention of multidrug-resistant tuberculosis in contacts. A cost-effectiveness analysis. Am J Respir Crit Care Med. 2015;192(2):229–37.

Chang KC, Yew WW, Chan RCY. Rapid assays for fluoroquinolone resistance in Mycobacterium tuberculosis: a systematic review and meta-analysis. J Antimicrob Chemother. 2010;65(8):1551–61.

Sotgiu G, Migliori GB. Facing multi-drug resistant tuberculosis. Pulm Pharmacol Ther. 2015;32:144–8.

Jabeen K, Shakoor S, Chishti S, Ayaz A, Hasan R. Fluoroquinolone-resistant Mycobacterium tuberculosis, Pakistan, 2005–2009. Emerg Infect Dis. 2011;17(3):566.

Van Der Heijden Y, Maruri F, Holt E, Mitchel E, Warkentin J, Sterling T. A comparison of interview methods to ascertain fluoroquinolone exposure before tuberculosis diagnosis. Epidemiol Infect. 2015;143(5):960–5.

Ignatyeva O, Balabanova Y, Nikolayevskyy V, Koshkarova E, Radiulyte B, Davidaviciene E, et al. Resistance profile and risk factors of drug resistant tuberculosis in the Baltic countries. Tuberculosis. 2015;95(5):581–8.

Mayer C, Takiff H. The molecular genetics of fluoroquinolone resistance in Mycobacterium tuberculosis.  Molecular Genetics of Mycobacteria, Second Edition: American Society of Microbiology; 2014. p. 455–78.

Aubry A, Veziris N, Cambau E, Truffot-Pernot C, Jarlier V, Fisher LM. Novel gyrase mutations in quinolone-resistant and-hypersusceptible clinical isolates of Mycobacterium tuberculosis: functional analysis of mutant enzymes. Antimicrob Agents Chemother. 2006;50(1):104–12.

Bernard C, Veziris N, Brossier F, Sougakoff W, Jarlier V, Robert J, et al. Molecular diagnosis of fluoroquinolone resistance in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2015;59(3):1519–24.

Bernard C, Aubry A, Chauffour A, Brossier F, Robert J, Veziris N. In vivo Mycobacterium tuberculosis fluoroquinolone resistance emergence: a complex phenomenon poorly detected by current diagnostic tests. J Antimicrob Chemother. 2016;71(12):3465–72.

Chen J, Peng P, Du Y, Ren Y, Chen L, Rao Y, et al. Early detection of multidrug-and pre-extensively drug-resistant tuberculosis from smear-positive sputum by direct sequencing. BMC Infect Dis. 2017;17(1):300.

Tavakoli A. Incidence and prevalence of tuberculosis in Iran and neighboring countries. Zahedan J Res Med Sci. 2017;19(7):e9238.

Bakuła Z, Napiórkowska A, Kamiński M, Augustynowicz-Kopeć E, Zwolska Z, Bielecki J, et al. Second-line anti-tuberculosis drug resistance and its genetic determinants in multidrug-resistant Mycobacterium tuberculosis clinical isolates. J Microbiol Immunol Infect. 2016;49(3):439–44.

Askarizadeh F, Aryan E, Hashemy SI, Youssefi M, Farsiani H, Ghazvini K. Prevalence and molecular characterization of Mycobacterium tuberculosis resistance to aminoglycosides in the Northeast of Iran. Acta microbiologica Hellenica. 2019;64(3):59–66.

Organization WH. Guidelines for treatment of drug-susceptible tuberculosis and patient care. 2017.

Jain N, Agnihotri S. Extensively drug resistance (XDR) TB-is not always fatal; 2009.

Falzon D, Gandhi N, Migliori GB, Sotgiu G, Cox HS, Holtz TH, et al. Resistance to fluoroquinolones and second-line injectable drugs: impact on multidrug-resistant TB outcomes. Eur Respir J. 2013;42(1):156–68.

Arjomandzadegan M, Titov L, Farnia P, Owlia P, Ranjbar R, Sheikholeslami F, et al. Molecular detection of fluoroquinolone resistance-associated gyrA mutations in ofloxacin-resistant clinical isolates of Mycobacterium tuberculosis from Iran and Belarus. Int J Mycobacteriol. 2016;5(3):299–305.

Yin X, Yu Z. Mutation characterization of gyrA and gyrB genes in levofloxacin-resistant Mycobacterium tuberculosis clinical isolates from Guangdong Province in China. J Infect. 2010;61(2):150–4.

Moure R, Tudó G, Medina R, Vicente E, Caldito JM, Codina MG, et al. Detection of streptomycin and quinolone resistance in Mycobacterium tuberculosis by a low-density DNA array. Tuberculosis. 2013;93(5):508–14.

Singh P, Jain A, Dixit P, Prakash S, Jaiswal I, Venkatesh V, et al. Prevalence of gyrA and B gene mutations in fluoroquinolone-resistant and-sensitive clinical isolates of Mycobacterium tuberculosis and their relationship with MIC of ofloxacin. J Antibiotics. 2015;68(1):63.

Singhal R, Reynolds PR, Marola JL, Epperson LE, Arora J, Sarin R, et al. Sequence analysis of fluoroquinolone resistance-associated genes gyrA and gyrB in clinical Mycobacterium tuberculosis isolates from patients suspected of having multidrug-resistant tuberculosis in New Delhi, India. J Clin Microbiol. 2016;54(9):2298–305.

Sharma R, Singh BK, Kumar P, Ramachandran R, Jorwal P. Presence of Fluoroquinolone mono-resistance among drug-sensitive Mycobacterium tuberculosis isolates: an alarming trend and implications. Clin Epidemiol Global Health. 2019;7(3):363–6.

Kim H, Mok JH, Kang B, Lee T, Lee H-K, Jang HJ, et al. Trend of multidrug and fluoroquinolone resistance in Mycobacterium tuberculosis isolates from 2010 to 2014 in Korea: a multicenter study. Korean J Intern Med. 2019;34(2):344.

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(1):364–9.

Rigouts L, Miotto P, Schats M, Lempens P, Cabibbe A, Galbiati S, et al. Fluoroquinolone heteroresistance in Mycobacterium tuberculosis: detection by genotypic and phenotypic assays in experimentally mixed populations. Sci Rep. 2019;9(1):1–8.

Pasca MR, Guglierame P, Arcesi F, Bellinzoni M, De Rossi E, Riccardi G. Rv2686c-Rv2687c-Rv2688c, an ABC fluoroquinolone efflux pump in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2004;48(8):3175–8.

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BMC Infectious Diseases

Tập 20 Số 1

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