Whole Genome Sequencing Results Associated with Minimum Inhibitory Concentrations of 14 Anti-Tuberculosis Drugs among Rifampicin-Resistant Isolates of Mycobacterium Tuberculosis from Iran

Kardan-Yamchi, Jalil; Kazemian, Hossein; Battaglia, Simone; Abtahi, Hamidreza; Foroushani, Abbas Rahimi; Hamzelou, Gholamreza; Cirillo, Daniela María; Ghodousi, Arash; Feizabadi, Mohammad M.

doi:10.3390/jcm9020465

Cited by 21 publications

(9 citation statements)

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“…The furA gene is known as the negative regulator of the katG gene; so a mutation in the furA gene or the furA-katG intergenic region may develop a level of INH resistance by reducing the katG expression [ 10 ]. Some studies have not found mutations in this gene in INH-resistant isolates [ 31 , 34 ], and others reported a small number of mutations at low frequencies from 2.3 to 3.7% [ 9 , 32 ]. Few of them have decreased katG expression and may be associated with low-level resistance [ 10 ].…”

Section: Discussionmentioning

confidence: 99%

Detection and characterization of mutations in genes related to isoniazid resistance in Mycobacterium tuberculosis clinical isolates from Iran

et al. 2022

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Background The global rise in drug-resistant Mycobacterium tuberculosis ( M.tb ), and especially the significant prevalence of isoniazid (INH)-resistance constitute a significant challenge to global health. Therefore, the present study aimed to investigate mutations in prevalent gene loci—involved in INH-resistance phenotype—among M.tb clinical isolates from southwestern Iran. Methods Drug susceptibility testing (DST) was performed using the conventional proportional method on confirmed 6620 M.tb clinical isolates, and in total, 15 INH-resistant and 18 INH-susceptible isolates were included in the study. Fragments of six genetic loci most related to INH-resistance ( katG , inhA promoter, furA , kasA , ndh , oxyR-ahpC intergenic region) were PCR-amplified and sequenced. Mutations were explored by pairwise alignment with the M.tb H37Rv genome. Results The analysis of gene loci revealed 13 distinct mutations in INH-resistant isolates. 60% (n = 9) of the INH-resistant isolates had mutations in katG , with codon 315 predominately (53.3%, n = 8). Mutation at InhA − 15 was found in 20% (n = 3) of resistant isolates. 26.7% (n = 4) of the INH-resistant isolates had kasA mutations, of which G269S substitution was the most common (20%, n = 3). The percentage of mutations in furA , oxyR-ahpC and ndh was 6.7% (n = 1), 46.7% (n = 7), and 20% (n = 3), respectively. Of the mutations detected in ndh and oxyR-ahpC , 5 were also observed in INH-susceptible isolates. This study revealed seven novel mutations, four of which were exclusively in resistant isolates. Conclusions This study supports the usefulness of katG and inhA mutations as a predictive molecular marker for INH resistance. Co-detection of katG S315 and inhA -15 mutations identified 73.3% (11 out of 15 isolates) of INH-resistant isolates.

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Section: Discussionmentioning

confidence: 99%

Detection and characterization of mutations in genes related to isoniazid resistance in Mycobacterium tuberculosis clinical isolates from Iran

et al. 2022

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“…The growth of the bacteria was checked at every 2 h by taking the optical density (OD) at 595 nm, and cfu was determined by dilution plating. MIC determination was done based on the broth dilution assay as per the CLSI guidelines. , Initial cfu and cfu after 12 h of the culture were also determined. The fold proliferation of the bacteria was measured as the ratio of cfu at 12 h to cfu at 0 h. The MIC of PAA-Cnp, Cnp, Ag nanoparticles, and ciprofloxacin as antibiotic control were done for S. Typhi and E.…”

Section: Methodsmentioning

confidence: 99%

Nanoceria-Based Phospholipase-Mimetic Cell Membrane Disruptive Antibiofilm Agents

Khulbe

Karmakar

Ghosh

et al. 2020

ACS Appl. Bio Mater.

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Table of Contents Chemicals and Methods S3 Characterisation of nanoparticles S4 POPG hydrolysis by MALDI-Mass Analysis S7 Phosphatidylcholine hydrolysis UV-Vis assay scheme S8 UV-Visible analysis of p-Nitrophenylphosphatidyl choline (NPPC) hydrolysis S9 Kinetic parameters of NPPC hydrolysis S10 31 P NMR analysis of NPPC hydrolysis S11 XPS and HRTEM analysis of nanoparticles S12 Phosphatidylcholine hydrolysis by nanoceria S13 Confocal study of bacterial cell membrane disruption S13 CFU analysis of Salmonella growth and death curve S15 Confocal microscopy analysis of antibacterial activity S17 Study of pre-attachment phase of biofilm formation S18 Synthesis and characterization of FITC-tagged PAA-Cnp S18 Localization of PAA-Cnp inside biofilm S21 Dispersibility of nanozymes in solution S21 SEM images of biofilm formation/disruption S22 Comparison of biofilm inhibition using oxidase-mimetic S22 Antibacterial activity on various pathogenic bacteria S23 Antibiofouling studies on urinary catheters S28 MIC quantification and comparison S29 Cytotoxicity study against HeLa cells S30 S3 1. Chemicals Cerium chloride heptahydrate (CeCl3.7H2O) was purchased from Avra synthesis Pvt. Ltd. Cerric ammonium nitrate ((NH4)2Ce(NO3)6), ethylenediamine, hydrazine hydrate and ammonium hydroxide, 3-aminopropyltriethoxysilane used for various synthesis were purchased from Sisco Research Laboratories. Sodium polyacrylate and flouresceine isothiocynate used for preparing PAA-Cnp and FITC-tagged PAA-Cnp nanoparticles respectively were purchased from Sigma Aldrich. Trizma base and phosphotidyl choline (lecithin) used for hydrolysis assays were also purchased from Sigma Aldrich. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol and pnitrophenyl phosphorylcholine used for other hydrolysis assays were purchased from TCI Co. Ltd. and Alfa Aesar Fischer Scientific respectively. 2. Characterization Methods Scanning electron microscopy (SEM) and EDX spectrawere performed on a Carl-Zeiss Ultra 55, FEI Sirion UHR SEM and ESEM-Quanta instruments,respectively. Transmission electron microscopy (TEM), high resolution TEM (HRTEM), electron diffraction X-Ray (EDX) analysis and selected area electron diffraction (SAED) were recorded on JEOL transmission electron microscope operating at 200 kV after casting a drop of nanoparticle dispersion in acetone over Cu grid. Powder XRDwas recordedon Philips PANalytical X-ray diffractometer by using a Cu-Kα (1.5406 Å)radiation. X-ray photoelectron spectroscopy (XPS) was acquired on AXIS ULTRA, KRATOS ANALYTICAL, SHIMADAZU. Raman spectroscopy was performed on HORIBA JOBIN YVON LabRAM HR Raman spectrometer (532 nm laser. FT-IR spectra were recorded on aPerkinElmer FT-IR spectrometer. Zeta potential was measured usingMalvern Zetasizer Nano UK.UV-Vis absorption spectra were acquired onSHIMADAZU UV-2600 spectrophotometer. Fluorescence spectroscopy was measured by using HORIBA JOBIN YVON (Fluoromax-4 Spectrofluorometer) instrument. All the 31 P NMR spectra were recorded using AV400 and AV500 MHz Avance Bruker High Resolution...

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“…Whole Genome Sequencing (WGS) can overcome this limitation by providing more information about diversity of sublineages due to its very high discrimination power [ 4 , 5 ]. Genomic diversity affects virulence, transmissibility, host response, and the emergence of drug resistance [ 6 , 7 , 8 ]. This information on distribution of lineage of MTB across geographic areas has empowered the global TB response.…”

Section: Introductionmentioning

confidence: 99%

Geno-Spatial Distribution of Mycobacterium Tuberculosis and Drug Resistance Profiles in Myanmar–Thai Border Area

et al. 2020

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Worldwide, studies investigating the relationship between the lineage of Mycobacterium tuberculosis (MTB) across geographic areas has empowered the “End TB” program and understand transmission across national boundaries. Genomic diversity of MTB varies with geographical locations and ethnicity. Genomic diversity can also affect the emergence of drug resistance. In Myanmar, we still have limited genetic information about geographical, ethnicity, and drug resistance linkage to MTB genetic information. This study aimed to describe the geno-spatial distribution of MTB and drug resistance profiles in Myanmar–Thailand border areas. A cross-sectional study was conducted with a total of 109 sequenced isolates. The lineages of MTB and the potential associated socio-demographic, geographic and clinical factors were analyzed using Fisher’s exact tests. p value of statistically significance was set at < 0.05. We found that 67% of the isolates were lineage 1 (L1)/East-African-Indian (EAI) (n = 73), followed by lineage 2 (L2)/Beijing (n = 26), lineage 4 (L4)/European American (n = 6) and lineage 3 (L3)/Delhi/Central Asian (n = 4). “Gender”, “type of TB patient”, “sputum smear grading” and “streptomycin resistance” were significantly different with the lineages of MTB. Sublineages of L1, which had never been reported elsewhere in Myanmar, were detected in this study area. Moreover, both ethnicity and lineage of MTB significantly differed in distribution by patient location. Diversity of the lineage of MTB and detection of new sublineages suggested that this small area had been resided by a heterogeneous population group who actively transmitted the disease. This information on distribution of lineage of MTB can be linked in the future with those on the other side of the border to evaluate cross-border transmission.

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Whole Genome Sequencing Results Associated with Minimum Inhibitory Concentrations of 14 Anti-Tuberculosis Drugs among Rifampicin-Resistant Isolates of Mycobacterium Tuberculosis from Iran

Cited by 21 publications

References 56 publications

Detection and characterization of mutations in genes related to isoniazid resistance in Mycobacterium tuberculosis clinical isolates from Iran

Detection and characterization of mutations in genes related to isoniazid resistance in Mycobacterium tuberculosis clinical isolates from Iran

Nanoceria-Based Phospholipase-Mimetic Cell Membrane Disruptive Antibiofilm Agents

Geno-Spatial Distribution of Mycobacterium Tuberculosis and Drug Resistance Profiles in Myanmar–Thai Border Area

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