scholarly journals Whole genomic sequencing as a tool for diagnosis of drug and multidrug-resistance tuberculosis in an endemic region in Mexico

2019 ◽  
Author(s):  
Carlos Francisco Madrazo-Moya ◽  
Irving Cancino-Muñoz ◽  
Betzaida Cuevas-Cordoba ◽  
Vanessa Gonzalez-Covarrubias ◽  
Martín Barbosa-Amezcua ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Multidrug Resistance ◽  
Multidrug Resistant ◽  
Genetic Distances ◽  
Clinical Isolates ◽  
Epidemiological Surveillance ◽  
Drug Resistant ◽  
Second Line ◽  
Endemic Region ◽  
Phenotypic Profile

AbstractBackgroundWhole genome sequencing (WGS) has been proposed as a tool for diagnosing drug resistance in tuberculosis. However, reports of its effectiveness in endemic countries with important numbers of drug resistance are scarce. The goal of this study was to evaluate the effectiveness of this procedure in isolates from a tuberculosis endemic region in Mexico.MethodsWGS analysis was performed in 81 tuberculosis positive clinical isolates with a known phenotypic profile of resistance against first-line drugs (isoniazid, rifampin, ethambutol, pyrazinamide and streptomycin). Mutations related to drug resistance were identified for each isolate; drug resistant genotypes were predicted and compared with the phenotypic profile. Genotypes and transmission clusters based on genetic distances were also characterized.FindingsPrediction by WGS analysis of resistance against isoniazid, rifampicin, ethambutol, pyrazinamide and streptomycin showed sensitivity values of 84%, 96%, 71%, 75% and 29%, while specificity values were 100%, 94%, 90%, 90% and 98%, respectively. Prediction of multidrug resistance showed a sensitivity of 89% and specificity of 97%. Moreover, WGS analysis revealed polymorphisms related to second-line drug resistance, enabling classification of eight and two clinical isolates as pre- and extreme drug-resistant cases, respectively.Four lineages were identified in the population (L1, L2, L3 and L4). The most frequent of these was L4, which included 90% (77) of the isolates. Six transmission clusters were identified; the most frequent was TC6, which included 13 isolates with a L4.1.1 and a predominantly multidrug-resistant condition.ConclusionThe results illustrate the utility of WGS for establishing the potential for prediction of resistance against first and second line drugs in isolates of tuberculosis from the region. They also demonstrate the feasibility of this procedure for use as a tool to support the epidemiological surveillance of drug- and multidrug-resistant tuberculosis.

scholarly journals Evaluation of Pyrosequencing for Detecting Extensively Drug-Resistant Mycobacterium tuberculosis among Clinical Isolates from Four High-Burden Countries

2014 ◽  
Vol 59 (1) ◽  
pp. 414-420 ◽  
Author(s):  
Kanchan Ajbani ◽  
Shou-Yean Grace Lin ◽  
Camilla Rodrigues ◽  
Duylinh Nguyen ◽  
Francine Arroyo ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
The Philippines ◽  
Drug Susceptibility Testing ◽  
Clinical Isolates ◽  
Drug Resistant ◽  
Second Line ◽  
Additional Advantage ◽  
Content Type ◽  
Extensively Drug Resistant

ABSTRACTReliable molecular diagnostics, which detect specific mutations associated with drug resistance, are promising technologies for the rapid identification and monitoring of drug resistance inMycobacterium tuberculosisisolates. Pyrosequencing (PSQ) has the ability to detect mutations associated with first- and second-line anti-tuberculosis (TB) drugs, with the additional advantage of being rapidly adaptable for the identification of new mutations. The aim of this project was to evaluate the performance of PSQ in predicting phenotypic drug resistance in multidrug- and extensively drug-resistant tuberculosis (M/XDR-TB) clinical isolates from India, South Africa, Moldova, and the Philippines. A total of 187 archived isolates were run through a PSQ assay in order to identifyM. tuberculosis(via the IS6110marker), and to detect mutations associated with M/XDR-TB within small stretches of nucleotides in selected loci. The molecular targets includedkatG, theinhApromoter and theahpC-oxyRintergenic region for isoniazid (INH) resistance; therpoBcore region for rifampin (RIF) resistance;gyrAfor fluoroquinolone (FQ) resistance; andrrsfor amikacin (AMK), capreomycin (CAP), and kanamycin (KAN) resistance. PSQ data were compared to phenotypic mycobacterial growth indicator tube (MGIT) 960 drug susceptibility testing results for performance analysis. The PSQ assay illustrated good sensitivity for the detection of resistance to INH (94%), RIF (96%), FQ (93%), AMK (84%), CAP (88%), and KAN (68%). The specificities of the assay were 96% for INH, 100% for RIF, FQ, AMK, and KAN, and 97% for CAP. PSQ is a highly efficient diagnostic tool that reveals specific nucleotide changes associated with resistance to the first- and second-line anti-TB drug medications. This methodology has the potential to be linked to mutation-specific clinical interpretation algorithms for rapid treatment decisions.

Chalcone derivatives and their activities against drug-resistant cancers: An overview

2020 ◽  
Vol 20 ◽  
Author(s):  
Jiaqi Xiao ◽  
Meixiang Gao ◽  
Qiang Diao ◽  
Feng Gao
Keyword(s):  
Drug Resistance ◽  
Multidrug Resistance ◽  
Cancer Cells ◽  
Anticancer Agents ◽  
Rational Design ◽  
Multidrug Resistant ◽  
Drug Resistant ◽  
Chalcone Derivatives ◽  
Potential Activity ◽  
Defensive Mechanisms

: Drug resistance including multidrug resistance resulting from different defensive mechanisms in cancer cells is the leading cause of the failure about the cancer therapy, making it an urgent need to develop more effective anticancer agents. Chalcones, widely distributed in nature, could act on diverse enzymes and receptors in cancer cells. Accordingly, chalcone derivatives possess potential activity against various cancers including drug-resistant even multidrug-resistant cancer. This review outlines the recent development of chalcone derivatives with potential activity against drug-resistant cancers covering articles published between 2010 and 2020, so as to facilitate further rational design of more effective candidate.

scholarly journals Second-line anti-tuberculosis drug resistance and its genetic determinants in multidrug-resistant Mycobacterium tuberculosis clinical isolates

2016 ◽  
Vol 49 (3) ◽  
pp. 439-444 ◽  
Author(s):  
Zofia Bakuła ◽  
Agnieszka Napiórkowska ◽  
Michał Kamiński ◽  
Ewa Augustynowicz-Kopeć ◽  
Zofia Zwolska ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Second Line ◽  
Genetic Determinants

scholarly journals PO 8408 DETECTION OF EXTENSIVELY DRUG-RESISTANT TUBERCULOSIS AMONG MULTIDRUG-RESISTANT MYCOBACTERIUM TUBERCULOSIS CLINICAL ISOLATES IN BOTSWANA

2019 ◽  
Vol 4 (Suppl 3) ◽  
pp. A33.1-A33
Author(s):  
Tuelo Mogashoa ◽  
Lucy Mupfumi ◽  
Thato Iketleng ◽  
Pinkie Melamu ◽  
Nametso Kelentse ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Geographical Location ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Drug Resistant ◽  
Second Line ◽  
Hiv Status ◽  
Extensively Drug Resistant ◽  
Line Drug

BackgroundThe emergence and transmission of multidrug-resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis (Mtb) strains is a serious threat to tuberculosis control in Botswana. Early detection of drug-resistant isolates is critical to ensure optimal treatment and thereby improve treatment outcomes. The objective of this study was to determine the extent of second-line drug resistance among drug-resistant Mtb-isolates from Botswana.MethodsA total of 60 drug-resistant Mtb isolates received at Botswana National Tuberculosis Reference Laboratory between 2012 and 2013 were analysed. DNA was extracted from BD Mycobacterial Growth Indicator Tubes (MGIT) using GenoLyse DNA isolation kit (Hain Lifescience). Spoligotyping was done using a commercially available spoligotyping kit (Isogen Life Science). The spoligotype patterns were compared with existing patterns in the SITVIT2 Web database. GenoType MTBDRs assay (Hain Lifescience) was used for second-line drug susceptibility testing. Fisher’s exact test was used to test for association between drug resistance patterns and HIV status, lineage and geographical location.ResultsSeventeen distinct spoligotype patterns were detected amongst the 60 drug-resistant isolates. The most predominant lineages were Euro-American (58.3%), East Asian (25%) and Indo-Oceanic (15%). Fifty (83.3%) were MDR, 7 (11.7%) were resistant to fluoroquinolones (Pre-XDR) whereas 3 (5%) were resistant to both fluoroquinolones and second-line injectable drugs (XDR). Drug resistance profiles were significantly associated with Mtb lineage (p<0.001). There was no association between drug resistance profile and HIV status (p=0.057) and geographical location (p=0.372).ConclusionThis study highlights the importance of including second-line drug susceptibility testing in a testing algorithm in Botswana. The detection of XDR isolates among MDR-TB isolates highlights the ongoing evolution of resistance and the need for strengthened treatment regimens to improve treatment outcomes and to prevent the spread of these highly resistant strains. Second-line testing will be essential if the 9 month MDR regimen is used in Botswana.

Assessing the utility of three TaqMan probes for the diagnosis of tuberculosis and resistance to rifampin and isoniazid in Veracruz, México

2012 ◽  
Vol 58 (3) ◽  
pp. 318-325 ◽  
Author(s):  
Roberto Zenteno-Cuevas ◽  
Betzaida Cuevas-Cordoba ◽  
Antonio Enciso ◽  
Leonor Enciso ◽  
Aremy Cuellar
Keyword(s):  
Drug Resistance ◽  
Molecular Mechanisms ◽  
Rpob Gene ◽  
Diagnostic Value ◽  
Multidrug Resistant ◽  
Molecular Probes ◽  
Clinical Isolates ◽  
Drug Resistant ◽  
Taqman Probes ◽  
Resistant Tuberculosis

Mutations at codons 526 and 531 in the rpoB gene and at 315 in the katG gene are considered diagnostic markers for resistance to rifampin and isoniazid in tuberculosis. The aim of this study was to design and evaluate three TaqMan probes for the identification of these mutations in 138 respiratory samples positive for acid-fast bacilli, and 32 clinical isolates from a region with considerable levels of drug resistance. The specificities of the probes for the diagnosis of resistance to both drugs were 100%; however, the sensitivities were calculated to be 50% for isoniazid and 56% for rifampin. DNA sequencing of rpoB and katG; and the spoligotyping assay of the clinical isolates, confirmed the diversity of the mutations and the presence of 11 spoligotypes with a shared international type and eight unique spoligotypes. Analysis of the respiratory samples identified 22 (16%) as drug-resistant and 4 (3%) as multidrug-resistant tuberculosis. The diagnostic value of the TaqMan probes was compromised by the diversity of mutations found in the clinical isolates. This highlights the need for better understanding of the molecular mechanisms responsible for drug resistance prior to the use of molecular probes, especially in regions with significant levels of drug-resistant tuberculosis.

scholarly journals Frequent suboptimal thermocycler ramp rate usage negatively impacts MTBDRsl performance for second-line drug resistant tuberculosis diagnosis

2021 ◽  
Author(s):  
Brigitta Derendinger ◽  
Margaretha de Vos ◽  
Samantha Pillay ◽  
Rouxjeane Venter ◽  
John Metcalfe ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Multidrug Resistant ◽  
Drug Resistant ◽  
Second Line ◽  
Ramp Rate ◽  
Dilution Series ◽  
Resistant Tuberculosis ◽  
Assay Performance ◽  
Mdr Tb ◽  
Line Drug

Strengthening the detection of second-line drug-resistance is a key tuberculosis (TB) control priority. The performance of MTBDRplus, a multidrug-resistant (MDR)-TB assay is reduced when suboptimal ramp rates are used. We investigated ramp rate's effect on MTBDRsl; the most widely-used molecular second-line drug-resistant TB assay. We tested 52 smear-negative Xpert MTB/RIF Ultra-positive sputa and a Mycobacterium tuberculosis (Mtb) dilution series at manufacturer recommended (2.2 ° C/s) or most common suboptimal ramp rate (4.0 ° C/s; identified via an earlier survey). Mtb-complex DNA (TUB-band)-positivity, indeterminate rates, fluoroquinolone- and second-line injectable-resistance accuracy, banding differences and, separately, inter-reader variability were assessed. 39% of re-surveyed laboratories (5/13) did not use the manufacturer-recommended MTBDRsl ramp rate. On sputum, this ramp rate improved indeterminates vs. 4.0 ° C/s (0/52 vs. 7/51; p=0.006), false drug-resistance calls (0/104 vs. 6/102; p=0.013), and incorrect banding calls (0/1300 vs. 55/1275; p<0.001). Valid results (neither TUB negative, indeterminate, nor any false drug-resistance calls) (52/52 vs. 41/51; p=0.001) on sputa hence improved by +21% (95% CI: 8-34%) with optimal ramp rate usage. Suboptimal ramp rate increased banding call inter-reader variability [52/1300 (4%) vs. 34/1300 (3%); p=0.030] on sputa but not dilution series; highlighting the importance of using clinical specimens for assay performance evaluations. Suboptimal ramp rate contributes to poor MTBDRsl performance. Ramp rate correction will improve second line drug-resistant TB diagnoses. Laboratories must ensure the optimal manufacturer-recommended ramp rate is used.

scholarly journals Prevalence and Genetic Characterization of Second-Line Drug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis in Rural China

2013 ◽  
Vol 57 (8) ◽  
pp. 3857-3863 ◽  
Author(s):  
Yi Hu ◽  
Sven Hoffner ◽  
Linlin Wu ◽  
Qi Zhao ◽  
Weili Jiang ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Dna Sequences ◽  
Rural China ◽  
Multidrug Resistant ◽  
Drug Resistant ◽  
Second Line ◽  
Content Type ◽  
Extensively Drug Resistant ◽  
Line Drug

ABSTRACTThis study aimed to investigate the prevalence of resistance to second-line antituberculosis (anti-TB) drugs and its association with resistance-related mutations inMycobacterium tuberculosisisolated in China. In the present study, we collected 380 isolates from a population-based study in China and tested the drug susceptibility to first- and selected second-line drugs. These results were compared with polymorphisms in the DNA sequences of genes associated with drug resistance and MIC values of the studied second-line drugs. Of 43 multidrug-resistantM. tuberculosisisolates, 13 showed resistance to fluoroquinolones or injectable second-line drugs (preextensively drug-resistant TB [pre-XDR-TB]), and 4 were resistant to both and thus defined as extensively drug-resistant TB (XDR-TB). Age and previous TB therapy, including use of second-line drugs, were two independent factors associated with increased resistance to both first- and second-line drugs. Molecular analysis identified the most frequent mutations in the resistance-associated genes: D94G ingyrA(29.1%) and A1401G inrrs(30.8%). Meanwhile, all 4 XDR-TB isolates had a mutation ingyrA, and 3 of them carried the A1401G mutation inrrs. Mutations ingyrAandrrswere associated with high-level resistance to fluoroquinolones and the second-line injectable drugs. In addition to the identification of resistance-associated mutations and development of a rapid molecular test to diagnose the second-line drug resistance, it should be a priority to strictly regulate the administration of second-line drugs to maintain their efficacy to treat multidrug-resistant TB.

scholarly journals Analysis of drug resistance and mutation profiles in Mycobacterium tuberculosis isolates in a surveillance site in Beijing, China

2021 ◽  
Vol 49 (1) ◽  
pp. 030006052098493
Author(s):  
Jie Zhang ◽  
Yixuan Ren ◽  
Liping Pan ◽  
Junli Yi ◽  
Tong Guan ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Drug Testing ◽  
Multidrug Resistant ◽  
High Rate ◽  
Drug Resistant ◽  
Surveillance Site ◽  
Mdr Tb ◽  
Previously Treated Patients ◽  
Beijing China

Objective This study analyzed drug resistance and mutations profiles in Mycobacterium tuberculosis isolates in a surveillance site in Huairou District, Beijing, China. Methods The proportion method was used to assess drug resistance profiles for four first-line and seven second-line anti-tuberculosis (TB) drugs. Molecular line probe assays were used for the rapid detection of resistance to rifampicin (RIF) and isoniazid (INH). Results Among 235 strains of M. tuberculosis, 79 (33.6%) isolates were resistant to one or more drugs. The isolates included 18 monoresistant (7.7%), 19 polyresistant (8.1%), 28 RIF-resistant (11.9%), 24 multidrug-resistant (MDR) (10.2%), 7 pre-extensively drug-resistant (XDR, 3.0%), and 2 XDR strains (0.9%). A higher rate of MDR-TB was detected among previously treated patients than among patients with newly diagnosed TB (34.5% vs. 6.8%). The majority (62.5%) of RIF-resistant isolates exhibited a mutation at S531L in the DNA-dependent RNA polymerase gene. Meanwhile, 62.9% of INH-resistant isolates carried a mutation at S315T1 in the katG gene. Conclusion Our results confirmed the high rate of drug-resistant TB, especially MDR-TB, in Huairou District, Beijing, China. Therefore, detailed drug testing is crucial in the evaluation of MDR-TB treatment.

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