Genotypic characterization of drug resistant Mycobacterium tuberculosis in Quebec, 2002-2012

Spinato, Joanna; Boivin, Élyse; Bélanger-Trudelle, Émilie; Fauchon, Huguette; Tremblay, Cécile; Soualhine, Hafid

doi:10.1186/s12866-016-0786-4

Cited by 12 publications

(8 citation statements)

References 29 publications

(34 reference statements)

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“…Most of the isolates had S315T and S531L mutations in the katG and rpoB genes, respectively. This is in line with what was observed among MDR isolates by using not only line-probe assays (69.8 vs. 55.9–90.6% for S315T katG; 64.1 vs. 41.4–67.2% for S531L rpoB ) (Farooqi et al, 2012 ; Shubladze et al, 2013 ; Sharma et al, 2014 ; Javaid et al, 2016 ; Spinato et al, 2016 ) but also sequencing strategy (66–76% for S315T katG; 46.3–66% for S531L rpoB ) (Jou et al, 2005 ; Ali et al, 2011 ; Makadia et al, 2012 ; Jagielski et al, 2015 ; Unissa et al, 2015 ). Both these mutations are thought to be low fitness cost mutations, with no adverse effect on transmission capacity (Gagneux, 2009 ).…”

Section: Discussionsupporting

confidence: 85%

Evaluation of Genotype MTBDRplus and MTBDRsl Assays for Rapid Detection of Drug Resistance in Extensively Drug-Resistant Mycobacterium tuberculosis Isolates in Pakistan

et al. 2018

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Pakistan ranks 5th among the world's highest tuberculosis (TB) burden countries alongside the 6th among countries with the highest burden of drug-resistant TB, including multi-drug resistant (MDR)-TB. Methods for rapid and reliable drug susceptibility testing (DST) are prerequisite for the prompt institution of effective anti-TB treatment. The aim of this study was to evaluate the efficiency of Genotype MTBDRplus and MTBDRsl assays for the detection of MDR and (pre-) extensively drug-resistant (XDR-TB) isolates in Pakistan. The study included 47 pre-XDR and 6 XDR-TB isolates, recovered from 53 patients from Pakistan. Conventional DST was performed using the standard 1% proportion method on the Löwenstein-Jensen medium. For molecular determination of drug resistance, GenoType MTBDRplus and GenoType MTBDRsl assays (Hain Lifescience, Germany) were used. To evaluate discrepancies between conventional and molecular DST results, mutation profiling was performed by amplifying and sequencing seven genetic loci, i.e., katG, inhA, and mabA-inhA promoter, rpoB, gyrA, embB, rrs. The sensitivity of Genotype MTBDRplus was 71.7% for isoniazid (INH) and 79.2% for rifampicin (RIF). Sequence analysis revealed non-synonymous mutations in 93.3 and 27.3% of isolates phenotypically resistant to INH and RIF, respectively, albeit susceptible when tested by GenoType MTBDRplus. GenoType MTBDRsl had a sensitivity of 73.6, 64.7, 20, 25, and 100% for the detection of fluoroquinolones, ethambutol, kanamycin, amikacin, and capreomycin resistance, respectively. Upon sequencing, mutations were detected in 20, 77.8%, and all isolates phenotypically resistant to aminoglycosides, ethambutol, and fluoroquinolones, respectively, yet declared as susceptible with GenoType MTBDRsl. Low sensitivities seriously impede the large-scale application of the Genotype MTBDRplus and MTBDRsl assays. Unless further optimized, the currently available line-probe assays should rather be auxiliary to the conventional, phenotype-based methods in the detection of MDR- and XDR-TB in Pakistan.

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Section: Discussionsupporting

confidence: 85%

Evaluation of Genotype MTBDRplus and MTBDRsl Assays for Rapid Detection of Drug Resistance in Extensively Drug-Resistant Mycobacterium tuberculosis Isolates in Pakistan

et al. 2018

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“…In this study, we detected 32 new mutations. [38][39][40][41][42] In addition, 89 of the100mutations were found in only one strain, and the remaining 11 mutations, which contained 35 strains, were shared mutations. Mutations in the promoter region were associated with PZA resistance.…”

Section: Discussionmentioning

confidence: 99%

<p>Pyrazinamide Resistance and Mutation Patterns Among Multidrug-Resistant <em>Mycobacterium tuberculosis</em> from Henan Province</p>

Shi¹,

Su²,

Zheng³

et al. 2020

IDR

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This study was designed to identify the phenotypic and genotypic characteristics of pyrazinamide (PZA) resistance among multidrug-resistant Mycobacterium tuberculosis (MDR-TB) from Henan and to evaluate the efficacy of pncA, rpsA, and panD mutations in predicting PZA resistance. Materials and Methods: A total of 152 MDR strains were included in this study. The Bactec MGIT system was used to determine PZA susceptibility for all strains. The pncA, rpsA, and panD genes were sequenced to identify any mutations, and the sequences were then aligned with the sequence of standard strain H37Rv. Moreover, the correlations between PZA-resistant phenotypes and treatment outcomes were analysed. Results: Of the152 strains, 105 had a PZA-resistant phenotype, and 102 harboured the pncA mutation. The PZA resistance rate was higher in the strains with resistance to all four firstline drugs and those that were pre-extensively drug-resistant (pre-XDR) and extensively drug-resistant (XDR). A total of 100 different pncA mutation patterns were identified, including 80 point mutations and 20 insertions/deletions, and 32 new pncA mutation patterns were detected. In this study, 13 strains had multiple mutations. Of the11 PZA-resistant strains without pncA mutations, two harboured the rpsA mutation, and one harboured the panD mutation. With PZA susceptibility results as the reference, single-gene pncA sequencing had sensitivity of 89.52% and specificity of 89.36%. With the combination of rpsA and panD, the sensitivity increased to 92.38%, and the specificity remained the same. No significant differences were observed in the sputum smear/culture conversion rate between PZAresistant patients and PZA-sensitive patients. However, PZA resistance was related to the time to sputum smear/culture conversion (P = 0.018). Conclusion: The combination of pncA, rpsA, and panD was beneficial for the timely diagnosis of PZA resistance and could provide a laboratory basis for customizing treatment regimens for MDR-TB patients.

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“…Isolates were plated on Lowenstein-Jensen slants at 37°C. Deoxyribonucleic acid was extracted from representative colonies using the BioRobot M48 (QIAGEN, Hilden, Germany) [5]. Identification to species level was done using a polymerase chain reaction (PCR)-based genomic deletion analysis of regions of difference [6].…”

Section: Methodsmentioning

confidence: 99%

Indigenous Transmission of Mycobacterium africanum in Canada: A Case Series and Cluster Analysis

Chakravarti

Soualhine

Sharma

et al. 2019

Open Forum Infectious Diseases

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Mycobacterium africanum is an important cause of human tuberculosis and is found almost exclusively in West Africa. We identified a cluster of patients in Montreal, Canada, with M africanum disease that share identical genotypic signatures by mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing and a putative epidemiological link, thus providing evidence of possible local transmission of M africanum in Montreal over a 10-year period.

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Genotypic characterization of drug resistant Mycobacterium tuberculosis in Quebec, 2002-2012

Cited by 12 publications

References 29 publications

Evaluation of Genotype MTBDRplus and MTBDRsl Assays for Rapid Detection of Drug Resistance in Extensively Drug-Resistant Mycobacterium tuberculosis Isolates in Pakistan

Evaluation of Genotype MTBDRplus and MTBDRsl Assays for Rapid Detection of Drug Resistance in Extensively Drug-Resistant Mycobacterium tuberculosis Isolates in Pakistan

<p>Pyrazinamide Resistance and Mutation Patterns Among Multidrug-Resistant <em>Mycobacterium tuberculosis</em> from Henan Province</p>

Indigenous Transmission of Mycobacterium africanum in Canada: A Case Series and Cluster Analysis

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