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Vol. 16. Issue 2.
Pages 136-141 (March - April 2012)
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Vol. 16. Issue 2.
Pages 136-141 (March - April 2012)
Open Access
Characterization of gyrA and gyrB mutations and fluoroquinolone resistance in Mycobacterium tuberculosis clinical isolates from Hubei Province, China
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Jun Chena,b,¥, Zhifei Chenb,¥, Yuanyuan Lia, Wei Xiaa, Xi Chena, Tian Chena, Liping Zhouc, Bin Xua, Shunqing Xua,
Corresponding author
xuscience@hotmail.com

Corresponding author at: Hangkong Road 13, Wuhan City, China.
a Ministry of Education Key Laboratory of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
b Wuhan Tuberculosis Prevention and Treatment Center, Wuhan, China
c Provincial Tuberculosis Reference Laboratory, Hubei Provincial Center for Disease Prevention and Control, Wuhan, China
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Abstract
Objective

The study aimed to investigate gyrA and gyrB mutations in Mycobacterium tuberculosis (MTB) clinical strains from 93 patients with pulmonary tuberculosis in Hubei Province, China, and analyze the association between mutation patterns of the genes and ofloxacin resistance level.

Results

Among 93 MTB clinical isolates, 61 were ofloxacin-resistant by the proportion method, and 32 were ofloxacin-susceptible MDR-TB. No mutation in the gyrB gene was found in any MTB strains. In the 61 ofloxacin-resistant isolates, 54 mutations were observed in the gyrA gene. Only one mutation in the gyrA gene was found in ofloxacinsusceptible MDR-TB isolates. In this study, the mutation patterns of gyrA involved seven patterns of single codon mutation (A90V, S91P, S91T, D94N, D94Y, D94G or D94A) and two patterns of double codons mutation (S91P & D94H, S91P & D94A). The ofloxacin minimal inhibitory concentrations (MICs) of three patterns of single codon mutations in the gyrA gene (codons 94, 90 and 91) showed a statistically significant difference (p<0.0001).

Conclusions

The gyrA mutations at codons 90, 91 and 94 constitute the primary mechanism of fluoroquinolone resistance in MTB, and mutations at codon 91 in the gyrA gene may be associated with low-level resistance to ofloxacin.

Keywords:
Mycobacterium tuberculosis
Fluoroquinolones
DNA mutational analysis
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References
[1.]
World Health Organization.
Multidrug and extensively drugresistant TB (M/XDR-TB): 2010 global report on surveillance and response.
World Health Organization, (2010),
[2.]
World Health Organization.
Guidelines for the programmatic management of drug resistant tuberculosis.
World Health Organization, (2006),
[3.]
T.S. Huang, C.M. Kunin, Shin-Jung Lee, et al.
Trends in fluoroquinolone resistance of Mycobacterium tuberculosis complex in a Taiwanese medical centre: 1995–2003.
J Antimicrob Chemother, 56 (2005), pp. 1058-1062
[4.]
K. Jabeen, S. Shakoor, S. Chishti, et al.
Fluoroquinoloneresistant Mycobacterium tuberculosis, Pakistan, 2005–2009.
Emerg Infect Dis, 17 (2011), pp. 564-566
[5.]
R. Shi, J. Zhang, C. Li, et al.
Emergence of ofloxacin resistance in Mycobacterium tuberculosis clinical isolates from China as determined by gyrA mutation analysis using denaturing high-pressure liquid chromatography and DNA sequencing.
J Clin Microbiol, 44 (2006), pp. 4566-4568
[6.]
J.C. Johnston, N.C. Shahidi, M. Sadatsafavi, et al.
Treatment outcomes of multidrug-resistant tuberculosis: a systematic review and meta-analysis.
[7.]
S.H. Gillespie, O. Billington.
Activity of moxifloxacin against mycobacteria.
J Antimicrob Chemother, 44 (1999), pp. 393-395
[8.]
C.A. Peloquin, D.J. Hadad, L.P. Molino, et al.
Population pharmacokinetics of levofloxacin, gatifloxacin, and moxifloxacin in adults with pulmonary tuberculosis.
Antimicrob Agents Chemother, 52 (2008), pp. 852-857
[9.]
C. Xu, B.N. Kreiswirth, S. Sreevatsan, et al.
Fluoroquinolone resistance associated with specific gyrase mutations in clinical isolates of multidrug-resistant Mycobacterium tuberculosis.
J Infect Dis, 174 (1996), pp. 1127-1130
[10.]
World Health Organization.
Guidelines for drug susceptibility testing for second-line anti-tuberculosis drugs for DOST-plus.
World Health Organization, (2001),
[11.]
A. Martin, J.C. Palomino, F. Portaels.
Rapid detection of ofloxacin resistance in Mycobacterium tuberculosis by two lowcost colorimetric methods: resazurin and nitrate reductase assays.
J Clin Microbiol, 43 (2005), pp. 1612-1616
[12.]
Z. Cui, J. Wang, J. Lu, et al.
Association of mutation patterns in gyrA/B genes and ofloxacin resistance levels in Mycobacterium tuberculosis isolates from East China in 2009.
BMC Infect Dis, 11 (2011), pp. 78
[13.]
I. Mokrousov, T. Otten, O. Manicheva, et al.
Molecular characterization of ofloxacin-resistant Mycobacterium tuberculosis strains from Russia.
Antimicrob Agents Chemother, 52 (2008), pp. 2937-2939
[14.]
A. Von Groll, A. Martin, P. Jureen, et al.
Fluoroquinolone resistance in Mycobacterium tuberculosis and mutations in gyrA and gyrB.
Antimicrob Agents Chemother, 53 (2009), pp. 4498-4500
[15.]
J.Y. Wang, L.N. Lee, H.C. Lai, et al.
Fluoroquinolone resistance in Mycobacterium tuberculosis isolates: associated genetic mutations and relationship to antimicrobial exposure.
J Antimicrob Chemother, 59 (2007), pp. 860-865
[16.]
X. Yin, Z. Yu.
Mutation characterization of gyrA and gyrB genes in levofloxacin-resistant Mycobacterium tuberculosis clinical isolates from Guangdong Province in China.
J Infect, 61 (2010), pp. 150-154
[17.]
A.F. Cheng, W.W. Yew, E.W. Chan, et al.
Multiplex PCR amplimer conformation analysis for rapid detection of gyrA mutations in fluoroquinolone-resistant Mycobacterium tuberculosis clinical isolates.
Antimicrob Agents Chemother, 48 (2004), pp. 596-601
[18.]
V. Jarlier, H. Nikaido.
Mycobacterial cell wall: structure and role in natural resistance to antibiotics.
FEMS Microbiol Lett, 123 (1994), pp. 11-18
[19.]
I. Escribano, J.C. Rodriguez, B. Llorca, et al.
Importance of the efflux pump systems in the resistance of Mycobacterium tuberculosis to fluoroquinolones and linezolid.
Chemotherapy, 53 (2007), pp. 397-401
[20.]
A.K. Gupta, V.M. Katoch, D.S. Chauhan, et al.
Microarray analysis of efflux pump genes in multidrug-resistant Mycobacterium tuberculosis during stress induced by common antituberculous drugs.
Microb Drug Resist, 16 (2010), pp. 21-28
[21.]
T. Kocagoz, C.J. Hackbarth, I. Unsal, et al.
Gyrase mutations in laboratory-selected, fluoroquinolone-resistant mutants of Mycobacterium tuberculosis H37Ra.
Antimicrob Agents Chemother, 40 (1996), pp. 1768-1774
[22.]
Z. Sun, J. Zhang, X. Zhang, et al.
Comparison of gyrA gene mutations between laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis strains and clinical isolates.
Int J Antimicrob Agents, 31 (2008), pp. 115-121
[23.]
Y. Hauck, M. Fabre, G. Vergnaud, et al.
Comparison of two commercial assays for the characterization of rpoB mutations in Mycobacterium tuberculosis and description of new mutations conferring weak resistance to rifampicin.
J Antimicrob Chemother, 64 (2009), pp. 259-262
[24.]
S.Y. Kim, Y.J. Park, W.I. Kim, et al.
Molecular analysis of isoniazid resistance in Mycobacterium tuberculosis isolates recovered from South Korea.
Diagn Microbiol Infect Dis, 47 (2003), pp. 497-502
[25.]
T. Lu, X. Zhao, K. Drlica.
Gatifloxacin activity against quinolone-resistant gyrase: allele-specific enhancement of bacteriostatic and bactericidal activities by the C-8-methoxy group.
Antimicrob Agents Chemother, 43 (1999), pp. 2969-2974
[26.]
T. Lu, X. Zhao, X. Li, et al.
Enhancement of fluoroquinolone activity by C-8 halogen and methoxy moieties: action against a gyrase resistance mutant of Mycobacterium smegmatis and a gyrase-topoisomerase IV double mutant of Staphylococcus aureus.
Antimicrob Agents Chemother, 45 (2001), pp. 2703-2709
[27.]
Y. Dong, X. Zhao, B.N. Kreiswirth, et al.
Mutant prevention concentration as a measure of antibiotic potency: studies with clinical isolates of Mycobacterium tuberculosis.
Antimicrob Agents Chemother, 44 (2000), pp. 2581-2584
[28.]
J. Poissy, A. Aubry, C. Fernandez, et al.
Should moxifloxacin be used for the treatment of extensively drug-resistant tuberculosis? An answer from a murine model.
Antimicrob Agents Chemother, 54 (2010), pp. 4765-4771

Both authors contributed equally to this work.

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