Genotyping and molecular characteristics of multidrug-resistant Mycobacterium tuberculosis isolates from China - 01/03/15

Doi : 10.1016/j.jinf.2014.11.008

Zhijian Zhang ^a,^b,^{^{e
These authors contributed equally to this work.}}, Jie Lu ^c,^{^{e
These authors contributed equally to this work.}}, Min Liu ^d,^{^{e
These authors contributed equally to this work.}}, Yufeng Wang ^b, Geping Qu ^a, Hongxia Li ^a, Jichun Wang ^b, Yu Pang ^a,^b,^⁎ , Changting Liu ^a,^⁎ , Yanlin Zhao ^b,^⁎
^a Respiratory Diseases Department of Nanlou, Chinese People's Liberation Army General Hospital, Beijing, China
^b National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China
^c 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
^d Liaoning Provincial Center for Disease Control and Prevention, Shenyang, China

^∗Corresponding author. National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Chang Bai Road, Changping District, Beijing 102206, China. Tel./fax: +86 10 5890 0779.^∗∗Corresponding author. National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Chang Bai Road, Changping District, Beijing 102206, China. Tel./fax: +86 10 5890 0777.^∗∗∗Corresponding author. Respiratory Department of Nanlou, Chinese People's Liberation Army General Hospital, 28# Fuxing Road, Haidian District, Beijing 100853, China. Tel./fax: +86 10 6687 6272.

Summary

Objectives

The aim of this study was to explore the population structure of multidrug-resistant (MDR) tuberculosis strains and distribution of resistance-associated nucleotide alteration among the different genotype MDR strains in China.

Methods

The genotypes of 376 MDR strain were analyzed by 15-loci MIRU-VNTR and RD105 deletion-targeted multiplex PCR (DTM-PCR) method. In addition, all the MDR isolates were sequenced for genetic mutations conferring rifampicin (rpoB) and isonizid resistance (katG, inhA and oxyR-ahpC).

Results

Among the 376 MDR isolates, 261 (69.4%) belonged to Beijing genotype, including 177 modern Beijing strains (67.8%) and 84 ancient Beijing (32.2%) strains. The percentages of streptomycin-resistant, kanamycin-resistant, pre-XDR and XDR TB in modern Beijing genotype were significantly lower than ancient genotype (P < 0.05). The Beijing MDR strains had significantly higher proportions of ofloxacin-resistant and pre-XDR isolates than non-Beijing strains (P < 0.01). In addition, the clustering rate of modern Beijing strains was significantly higher than that of ancient Beijing strains (46.3% vs. 11.9%, P < 0.01). 94.7% and 79.3% of MDR isolates harbored genetic mutations conferring rifampicin and isonizid resistance, respectively, and the most prevalent mutation was located in codon rpoB531 and katG315. In addition, the rpoB531 and katG mutation were more frequently observed among Beijing genotype strains than non-Beijing strains, while non-Beijing genotype showed stronger association with isolates lacking mutation in rifampicin resistance determination region (P < 0.05).

Conclusions

Our findings demonstrated that ancient Beijing MDR strains were associated with drug resistance, while modern Beijing MDR strains were more likely to be clustered.

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Highlights

•	We investigated the genetic diversity of multidrug-resistant tuberculosis strains in China.
•	The ancient Beijing MDR strains were associated with drug resistance.
•	The modern Beijing MDR strains were more likely to be clustered.

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Keywords : Mycobacterium tuberculosis, Multidrug-resistant, Genotyping

Plan

Introduction

Materials and methods

Bacterial strains and culture conditions

Genomic DNA extraction

Association between rpoB mutations and genotypes

Association between INH resistance-related mutations and genotypes

Discussion

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Vol 70 - N° 4

P. 335-345 - avril 2015 Retour au numéro

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Genotyping and molecular characteristics of multidrug-resistant Mycobacterium tuberculosis isolates from China - 01/03/15

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