Multidrug-resistant Mycobacterium Tuberculosis Strains Research Articles

Model-based impact evaluation of new tuberculosis vaccines in aging populations under different modeling scenarios: the case of China.

The slow descent in TB burden, the COVID-19 pandemic, along with the rise of multidrug-resistant strains of Mycobacterium tuberculosis, seriously threaten TB control and the goals of the End TB strategy. To fight back, several vaccine candidates are under development, with some of them undergoing the phases 2B and 3 of the development pipeline. The impact of these vaccines on the general population needs to be addressed using disease-transmission models, and, in a country like China, which last year ranked third in number of cases worldwide, and where the population is aging at a fast pace, the impact of TB vaccination campaigns may depend heavily upon the age of targeted populations, the mechanistic descriptions of the TB vaccines and the coupling between TB dynamics and demographic evolution. In this work, we studied the potential impact of a new TB vaccine in China targeting adolescents (15-19 y.o.) or older adults (60-64 y.o.), according to varying vaccine descriptions that represent reasonable mechanisms of action leading to prevention of disease, or prevention of recurrence, each of them targetting specific routes to TB disease. To measure the influence of the description of the coupling between transmission dynamics and aging in TB transmission models, we explored two different approaches to compute the evolution of the contact matrices, which relate to the spreading among different age strata. Our findings highlight the dependence of model-based impact estimates on vaccine profiles and the chosen modeling approach for describing the evolution of contact matrices. Our results also show, in line with previous modeling works, that older adult vaccination is a suitable option in China to reduce the incidence of TB as long as the vaccine is able to protect already exposed individuals. This study underscores the importance of considering vaccine characteristics and demographic dynamics in shaping TB control strategies. In this sense, older adult vaccination emerges as a promising avenue for mitigating TB transmission in China but also remarks the need for tailored intervention strategies aligned with demographic trends.

1954. Dynamic PET facilitated Modeling and Novel Antibiotic Regimens for Tuberculous Meningitis due to Multidrug-Resistant Strains

1954. Dynamic PET facilitated Modeling and Novel Antibiotic Regimens for Tuberculous Meningitis due to Multidrug-Resistant Strains

Characterization of Mutations in Genes Related to Rifampicin and Isoniazid Resistance in Multidrug-resistant Mycobacterium tuberculosis Strains from Hangzhou, China

Characterization of Mutations in Genes Related to Rifampicin and Isoniazid Resistance in Multidrug-resistant Mycobacterium tuberculosis Strains from Hangzhou, China

Characterisation and development of histopathological lesions in a guinea pig model of Mycobacterium tuberculosis infection.

Tuberculosis (TB) remains a very significant infectious disease worldwide. New vaccines and therapies are needed, even more crucially with the increase of multi-drug resistant Mycobacterium tuberculosis strains. Preclinical animal models are very valuable for the development of these new disease control strategies. Guinea pigs are one of the best models of TB, sharing many features with the pathology observed in human TB. Here we describe the development of TB lesions in a guinea pig model of infection. We characterise the granulomatous lesions in four developmental stages (I-IV), using histopathological analysis and immunohistochemical (IHC) techniques to study macrophages, T cells, B cells and granulocytes. The granulomas in the guinea pigs start as aggregations of macrophages and few heterophils, evolving to larger lesions showing central caseous necrosis with mineralisation and abundant acid-fast bacilli, surrounded by a rim of macrophages and lymphocytes in the outer layers of the granuloma. Multinucleated giant cells are very rare and fibrotic capsules are not formed in this animal model.

Identification of novel antimicrobial compounds targeting Mycobacterium tuberculosis shikimate kinase using in silico hierarchical structure-based drug screening

The development of new anti-TB drugs to prevent the spread of multidrug-resistant Mycobacterium tuberculosis (Mtb) strains is imperative. Mtb shikimate kinase (MtSK) was selected as the target protein to screen for new anti-TB drugs. We performed hierarchical in silico screening using a library of 154,118 compounds to search for novel compounds that could bind to the active site of MtSK. The growth-inhibitory effects of the candidate compounds on Mycobacterium smegmatis were evaluated in vitro. Nine of the 11 candidate compounds exhibited inhibitory effects against mycobacteria in vitro. The inhibitory activity of Compound 2 (IC50 = 1.39 μM) was higher than that of isoniazid, the first-line drug for TB treatment. Moreover, Compound 2 did not exhibit toxicity against mammalian cells and Escherichia coli. Molecular dynamics simulations using the MtSK-Compound 2 complex structure in a timeframe of 100 ns suggested that Compound 2 could stably bind to MtSK. The binding free energy of Compound 2 was estimated to be −37.96 kcal/mol using the MM/PBSA method, demonstrating that Compound 2 can stably bind to MtSK. These in silico and in vitro results indicated that Compound 2 is a promising hit compound for the development of novel anti-TB drugs.

Epidemiological manifestations of tuberculosis infection in the Omsk region: dynamics and trends

Background. Tuberculosis (TB) infection remains relevant as one of the leading public health problems in Russia.The aim. To characterize the dynamics and trends of epidemiological manifestations of TB infection in the Omsk region.Materials and methods. An observational descriptive-evaluative epidemiological study was carried out in the Omsk region from 2009 to 2021. The data available in the Federal statistical observation forms NN 7, 8, and 33, and results of the bacteriological study of patients with respiratory TB were analyzed.Results. In the Omsk region, an improvement in the epidemiological situation was observed from 2009 to 2021. It was associated with a decrease in TB prevalence by 3.7 times (from 325.6 to 86.7), mortality – by 5.8 times (from 21.1 to 3.6), incidence – by 2.7 times (from 130.7 to 48.4 per 100,000). At the same time, proportion of microscopy/culture-positive patients infected with multidrug-resistant Mycobacterium tuberculosis strains increased from 9.2 to 29.8 % among all patients, and from 15.5 to 30.6 % among newly diagnosed patients. There was a trend towards an increase in the number of cases with primary extensive drug resistance of M. tuberculosis. The incidence of tuberculosis associated with HIV infection has increased 10 times and reached 15.6 per 100,000 population.Conclusions. In the Omsk region, there is a change in the structure of M. tuberculosis strains with a predominance of multiple and extensive drug resistance along with decrease in TB incidence and mortality. New approaches are needed to organize the system of epidemiological surveillance and control of TB infection.

Recent Advances in Anti-Tuberculosis Drug Discovery Based on Hydrazide-Hydrazone and Thiadiazole Derivatives Targeting InhA.

Tuberculosis is an extremely serious problem of global public health. Its incidence is worsened by the presence of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis. More serious forms of drug resistance have been observed in recent years. Therefore, the discovery and/or synthesis of new potent and less toxic anti-tubercular compounds is very critical, especially having in mind the consequences and the delays in treatment caused by the COVID-19 pandemic. Enoyl-acyl carrier protein reductase (InhA) is an important enzyme involved in the biosynthesis of mycolic acid, a major component of the M. tuberculosis cell wall. At the same time, it is a key enzyme in the development of drug resistance, making it an important target for the discovery of new antimycobacterial agents. Many different chemical scaffolds, including hydrazide hydrazones and thiadiazoles, have been evaluated for their InhA inhibitory activity. The aim of this review is to evaluate recently described hydrazide-hydrazone- and thiadiazole-containing derivatives that inhibit InhA activity, resulting in antimycobacterial effects. In addition, a brief review of the mechanisms of action of currently available anti-tuberculosis drugs is provided, including recently approved agents and molecules in clinical trials.

Molecular Design and In-Silico Analysis of Trisubstituted Benzimidazole Derivatives as Ftsz Inhibitor

Tuberculosis (TB) is the fastest spreading infectious disease and one of the top ten diseases that kill millions of people annually. The rapid spread of a multidrug-resistant strain of Mycobacterium tuberculosis leads to multidrug-resistance tuberculosis (MDR-TB), which is very difficult to treat. Filament temperature-sensitive protein ring-Z (Ftsz) protein could be the best target to inhibit bacterial cytokinesis. This research is conducted to predict the antitubercular activity of trisubstituted benzimidazole derivatives targeting FtsZ protein by an in-silico approach (molecular docking, pharmacokinetic parameter, drug likeliness, toxicity prediction, and biological activity prediction). Amine and aldehyde substitutions are used as primary scaffolds to design 20 trisubstituted benzimidazole derivatives for molecular docking. AutoDock vina v.1.2.0 software was used to predict the binding interaction between ligand and receptor (FtsZ, PDB ID : 1RQ7). The drug-likeliness properties and toxicity of ligands were predicted from SwissADMET and ToxiM web servers, respectively. Compound A15 (2,3,5,6-tetrafluoro-N1-{6-fluoro-5-[4-(1H-imidazole-1-yl) phenoxy]-1H-1,3-benzodiazol-2-yl} benzene-1,4-diamine) showed the best binding energy (ΔG = −10.2 kcal/mol/) along with four hydrogen bond interactions (GLY107, PHE180, ASP 184). Similarly, compounds A19 and A20 have the best binding score of −9.8 kcal/mol, with excellent pharmacokinetic parameters. It is found that the binding energy of all ligands (ΔG = −8.0 to −10.2 kcal/mol) is better than the reference compound Moxifloxacin (ΔG = −7.7 kcal/mol). None of the ligands violate Lipinski’s rule, but all ligands’ toxicity is slightly high (>0.8 score). It is reported that the amine-substituted benzimidazole derivatives have better binding energy than the aldehyde substitution. Therefore, it is concluded that compounds A19 and A20 can be the best candidate as Ftsz protein inhibitors but an in-vitro animal study and toxicity study are necessary to validate these data.

An In-Silico Reverse Vaccinology Approach in Predicting Vaccine Candidate Against Mycobacterium tuberculosis CCDC5180

A quarter of the world’s population is thought to be latently infected with Mycobacterium tuberculosis, creating a vast pool of possible future illnesses. Combined with the rising prevalence of multi-drug resistant Mycobacterium tuberculosis (Mtb) strains, TB poses one of the biggest challenges for world health, and the TB epidemic can only be eradicated with a new and more effective vaccine strategy. Since the Bacille Calmette-Guérin (BCG) vaccine is the only one used to treat tuberculosis (TB), Reverse vaccine technology aims to accelerate the development of subunit vaccines by identifying specific proteins in a pathogenic bacterial proteome that may be protective antigens. This approach was employed on four extracellular proteins namely Secreted fibronectin-binding protein C antigen 85-C, PE-PGRS family protein, Invasion protein RipA, and Invasion protein RipB. The shortlisted proteins were subjected to B-cell, CTL, and HTL epitope prediction. To further identify the most promising epitopes, specialized filtering techniques were employed to narrow down CTL epitopes that were non-allergenic, non-toxic, and antigenic as well as B-cell epitopes that produced antibodies. Similarly, HTL epitopes that generate IFN- γ but not IL-10 were selected. The analysis revealed invasion protein RipA as a potential candidate for Mycobacterium tuberculosis CCDC5180 immunization.

Design, Synthesis, Characterization of Novel Sulfathiazole Derivatives and their in silico and in vitro Analysis against Multidrug-Resistance Tuberculosis using Docking Studies

In this study, seven novel sulfathiazole derivatives were synthesized from sulfathiazole using different substituted aldehydes and characterized by IR, NMR and LC-MS analysis. Using molecular docking and toxicity prediction, all the seven novel sulfathiazole derivatives (Mol-14, Mol-15, Mol-21, Mol-27, Mol-36, Mol-39 and Mol-43) were virtually screened from generated 70 compounds and assessed their effectiveness against multidrug resistant Mycobacterium tuberculosis (MDR-TB). The Inha protein of TB were performed and all the compounds found to have good docking scores in the range of -7.2 to -9.1 Kcal/mol. Compound, 4-(4-oxo-2-phenyl-1,3-thiazolidin-3-yl)-N-(1,3-thiazol-2-yl)benzene-1- sulfonamide (Mol-27) shown to inhibit the MDR-TB and wild-type TB strain with an MIC value of 1 μg/mL and 0.25, respectively. The standard sulfathiazole and isoniazid were compared to the minimal inhibitory concentration (MIC) of the synthesized Mol-14, Mol-15, Mol-21, Mol-27, Mol-36, Mol-39 and Mol-43 new sulfathiazole derivatives. Based on the results, these compounds shows promising activity against MDR-TB.

Dithiocarbamates and dithiocarbonates containing 6-nitrosaccharin scaffold: Synthesis, antimycobacterial activity and in silico target prediction using ensemble docking-based reverse virtual screening

Tuberculosis is a curable disease that costs the lives of 1.5 million people per year. It continues to pose a threat to global public health owing to the prevalence of multi-drug resistant (MDR) Mycobacterium tuberculosis strains which make its eradication a continuous challenge. To explore novel and effective antimycobacterial agents, a series of dithiocarbamates/dithiocarbonate-nitrosaccharin hybrids were prepared, characterized by spectral analyses, and assessed for their antimycobacterial activity against M.tuberculosis strain H37Rv. Some compounds showed activity with a minimum inhibitory concentration (MIC) of 0.45–6.09 µg/mL. Reverse ensemble docking against nine putative targets with fifteen different crystal structures was conducted to understand the mechanism of action of the most potent compound 6g. The in silico studies suggested enoyl ACP-reductase (InhA) and polyketide synthase 13 (Pks13) as possible targets for compound 6g.

Modified gold nanoparticle colorimetric probe-based biosensor coupled with allele-specific PCR for rapid detection of G944C mutation associated with isoniazid resistance

The emergence of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis (MTB) makes tuberculosis (TB) difficult to control and treat. Therefore, rapid and reliable detection of MDR-TB within the community plays a significant role. The aim of this study was to investigate the use of modified gold nanoparticle (AuNP) colorimetric probe-based biosensor coupled with allele-specific PCR for the rapid detection of G944C mutation associated with isoniazid resistance in MTB. Based on the citrate reduction method, the AuNPs were synthesized and functionalized using thiol-modified oligonucleotides (AuNP-biosensor). The AuNP-biosensor method was compared to the gold standard in terms of analytical and clinical sensitivity and specificity, positive predictive value (PPV), negative predictive value (NPV), diagnostic odds ratio (DOR), and accuracy. The gold standard was defined as a positive result in sequencing.The AuNP-biosensor had 100% sensitivity and specificity for detection of G944C mutation. After allele-specific PCR amplification the results were carried out for <15 min with ready-to-use AuNP-biosensor. PPV, NPV, DOR and accuracy of this method were 100%, 100%, 309 and 100%, respectively. The method designed in this study can identify the desired mutation with high efficiency after initial amplification, either from the sputum sample or from isolated bacteria. By designing a panel of this method, different MTB resistance-associated mutations could be detected more rapidly.

Adenosine-Mimicking Derivatives of 3-Aminopyrazine-2-Carboxamide: Towards Inhibitors of Prolyl-tRNA Synthetase with Antimycobacterial Activity

Multidrug-resistant tuberculosis (MDR-TB) poses a significant threat to mankind and as such earned its place on the WHO list of priority pathogens. New antimycobacterials with a mechanism of action different to currently used agents are highly required. This study presents the design, synthesis, and biological evaluation of 3-acylaminopyrazine-2-carboxamides derived from a previously reported inhibitor of human prolyl-tRNA synthetase. Compounds were evaluated in vitro against various strains of mycobacteria, pathogenic bacteria, and fungi of clinical significance. In general, high activity against mycobacteria was noted, while the antibacterial and antifungal activity was minimal. The most active compounds were 4'-substituted 3-(benzamido)pyrazine-2-carboxamides, exerting MIC (Minimum Inhibitory Concentration) from 1.95 to 31.25 µg/mL. Detailed structure-activity relationships were established and rationalized in silico with regard to mycobacterial ProRS as a probable target. The active compounds preserved their activity even against multidrug-resistant strains of Mycobacterium tuberculosis. At the same time, they were non-cytotoxic against HepG2 human hepatocellular carcinoma cells. This project is the first step in the successful repurposing of inhibitors of human ProRS to inhibitors of mycobacterial ProRS with antimycobacterial activity.

Host cell transcriptomic response to the multidrug-resistant Mycobacterium tuberculosis clonal outbreak Beijing strain reveals its pathogenic features

ABSTRACT The upsurge of multidrug-resistant infections has rendered tuberculosis the principal cause of death among infectious diseases. A clonal outbreak multidrug-resistant triggering strain of Mycobacterium tuberculosis was identified in Kanchanaburi Province, labelled “MKR superspreader,” which was found to subsequently spread to other regions, as revealed by prior epidemiological reports in Thailand. Herein, we showed that the MKR displayed a higher growth rate upon infection into host macrophages in comparison with the H37Rv reference strain. To further elucidate MKR’s biology, we utilized RNA-Seq and differential gene expression analyses to identify host factors involved in the intracellular viability of the MKR. A set of host genes function in the cellular response to lipid pathway was found to be uniquely up-regulated in host macrophages infected with the MKR, but not those infected with H37Rv. Within this set of genes, the IL-36 cytokines which regulate host cell cholesterol metabolism and resistance against mycobacteria attracted our interest, as our previous study revealed that the MKR elevated genes associated with cholesterol breakdown during its growth inside host macrophages. Indeed, when comparing macrophages infected with the MKR to H37Rv-infected cells, our RNA-Seq data showed that the expression ratio of IL-36RN, the negative regulator of the IL-36 pathway, to that of IL-36G was greater in macrophages infected with the MKR. Furthermore, the MKR’s intracellular survival and increased intracellular cholesterol level in the MKR-infected macrophages were diminished with decreased IL-36RN expression. Overall, our results indicated that IL-36RN could serve as a new target against this emerging multidrug-resistant M. tuberculosis strain.

Total Synthesis and Biological Evaluation of Modified Ilamycin Derivatives.

Ilamycins/rufomycins are marine cycloheptapeptides containing unusual amino acids. Produced by Streptomyces sp., these compounds show potent activity against a range of mycobacteria, including multidrug-resistant strains of Mycobacterium tuberculosis. The cyclic peptides target the AAA+ protein ClpC1 that, together with the peptidases ClpP1/ClpP2, forms an essential ATP-driven protease. Derivatives of the ilamycins with a simplified tryptophane unit are synthesized in a straightforward manner. The ilamycin derivative 26 with a cyclic hemiaminal structure is active in the nM-range against several mycobacterial strains and shows no significant cytotoxicity. In contrast, derivative 27, with a glutamic acid at this position, is significantly less active, with MICs in the mid µM-range. Detailed investigations of the mode of action of 26 indicate that 26 deregulates ClpC1 activity and strongly enhances ClpC1-WT ATPase activity. The consequences of 26 on ClpC1 proteolytic activities were substrate-specific, suggesting dual effects of 26 on ClpC1-WT function. The positive effect relates to ClpC1-WT ATPase activation, and the negative to competition with substrates for binding to the ClpC1 NTD.

Novel diaryl ether derivatives as InhA inhibitors: Design, synthesis and antimycobacterial activity

A new series of triclosan (TCL)-mimicking diaryl ether derivatives 7–25 were synthesized and evaluated as inhibitors of enoyl acyl carrier protein reductase InhA enzyme. In addition, these derivatives were screened as inhibitors of drug-susceptible (DS), multidrug-resistant (MDR), and extensive drug-resistant (XDR) Mycobacterium tuberculosis (MTB) strains. Most compounds exihibted superior anti-TB activities and improved ClogP compared to TCL as a standard drug. The present work has led to the identification of compounds 14, 19 and 24 which possess remarkable activities against DS, MDR and XDR MTB strains with MIC values of 1.95, 3.9 and 15.63 µg/ml, respectively for compound 14, 1.95, 3.9 and 7.81 µg/ml, respectively for compound 19 and 0.98, 1.95 and 3.9 µg/ml, respectively for compound 24. Most compounds did not exhibit toxicity to HePG2 normal cell line. Compounds 14, 19 and 24, presenting the best MIC values, were further evaluated as inhibitors of InhA enzyme. They showed high binding affinities in the micromolar range with IC50 values of 1.33, 0.6, and 0.29 µM for compounds 14, 19, and 24, respectively. Furthermore, molecular docking approach was utilized to understand the difference in bioactivities between the new compounds. In particular, the results revealed strong binding interactions and high docking scores of compounds 14, 19 and 24, which could correlate with their high activities. Mainly, the molecular modelling study of compound 24 provides an excellent platform for understanding the molecular mechanism regarding InhA inhibition. Thus, compound 24 could be a lead compound for future development of new antitubercular drugs.

Decreasing trend of drug-resistant TB in Italy

BACKGROUND: TB caused by rifampicin-resistant (RR) and multidrug-resistant (MDR) Mycobacterium tuberculosis strains is a major concern to TB control globally. However, in the European Union, MDR-TB notifications among all bacteriologically confirmed TB cases with available drug susceptibility testing (DST) results decreased over the last years.METHODS: We conducted a retrospective analysis on DST results reported from 2011 to 2020 by 46 laboratories in 19 out of 20 regions in Italy in order to evaluate resistance trends to first- and second-line drugs in MDR/RR-TB strains isolated from Italian-born persons (IBPs) and foreign-born persons (FBPs).RESULTS: Of 23,972 M. tuberculosis strains examined (15,519 from FBPs and 8,453 from IBPs), MDR-TB decreased from 3.2% in 2011 to 2.2% in 2020. High MDR/RR-TB rates occurred mostly in FBPs from former Soviet Union countries. In 2017, a MDR/RR-TB increase was detected in FBPs from sub-Saharan Africa. MDR-TB strains showed consistent increase in resistance to pyrazinamide (PZA), slight increase in resistance to fluoroquinolones and a decrease in resistance to other drugs.CONCLUSION: While MDR/RR-TB cases slightly decreased, a worrisome increase of resistance to PZA and fluoroquinolones among MDR/RR-TB patients was seen. This implies that a fast and efficient diagnosis aligned with therapy is crucial for TB control.

RESISTANCE MECHANISMS TO FIRST LINE DRUG IN MYCOBACTERIUM TUBERCULOSIS: A REVIEW

The emergence of multidrug-resistant (MDR) Mycobacterium tuberculosis (Mtb) strains, identified as resistant to at least isoniazid and rifampin, these two drugs form the backbone of the first line drug which is used for tuberculosis (TB) treatment, unresponsive has hampered TB control. As a result of the nearly universal calculation of with half a million new cases of MDR/first line drug TB per year, it is important to keep the database up to date awareness of the processes that contribute to the emergence of MDR Mtb. This resistance is produced for a variety of reasons, including genetic, microbiological factors, non-adherence to treatment by patients and/or failures in therapy administration by some referrer medical centre for TB. This review offers a detailed summary of genetic mechanisms that lead to resistant to first line drug therapy used in management of TB as well as up-to-date information on some new aspects lead to such problem. Keywords: Mycobacterium tuberculosis, first line drug, anti-TB treatment, World Health Organization, isoniazid and rifampin Citation: Hassan JS, Al-Tamimi BJ, Abdulrahman TR, Jamal QW. Resistance mechanisms to first line drug in Mycobacterium tuberculosis: A review. Iraqi JMS. 2022; 20(1): 35-43. doi: 10.22578/IJMS.20.1.5

Antitubercular, Cytotoxicity, and Computational Target Validation of Dihydroquinazolinone Derivatives.

A series of 2,3-dihydroquinazolin-4(1H)-one derivatives (3a–3m) was screened for in vitro whole-cell antitubercular activity against the tubercular strain H37Rv and multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) strains. Compounds 3l and 3m with di-substituted aryl moiety (halogens) attached to the 2-position of the scaffold showed a minimum inhibitory concentration (MIC) of 2 µg/mL against the MTB strain H37Rv. Compound 3k with an imidazole ring at the 2-position of the dihydroquinazolin-4(1H)-one also showed significant inhibitory action against both the susceptible strain H37Rv and MDR strains with MIC values of 4 and 16 µg/mL, respectively. The computational results revealed the mycobacterial pyridoxal-5′-phosphate (PLP)-dependent aminotransferase (BioA) enzyme as the potential target for the tested compounds. In vitro, ADMET calculations and cytotoxicity studies against the normal human dermal fibroblast cells indicated the safety and tolerability of the test compounds 3k–3m. Thus, compounds 3k–3m warrant further optimization to develop novel BioA inhibitors for the treatment of drug-sensitive H37Rv and drug-resistant MTB.

Determination of genetic diversity of multidrug-resistant Mycobacterium tuberculosis strains in Turkey using 15 locus MIRU-VNTR and spoligotyping methods

ABSTRACT Tuberculosis (TB) remains the leading cause of deaths from infectious disease worldwide. Nowadays, the tendency of Mycobacterium tuberculosis complex (MTBC) to spread between continents due to uncontrolled migration movements shows that TB is a global health problem. The number of studies for the detection of MTBC strains’ epidemiological features in areas with TB spread risk using molecular-based methods such as spoligotyping and Mycobacterial Interspersed Repetitive Unit (MIRU) Variable Number Tandem Repeats (VNTR) at the clonal level is insufficient. In this study, it was aimed to determine the phylogenetic relationships of MTBC strains at the species level by spoligotyping and 15 locus MIRU-VNTR (MIRU-VNTR15) molecular methods of 96 multidrug-resistant (MDR) MTBC strains isolated from sputum samples of patients with a preliminary diagnosis of pulmonary TB or suspected contact history those sent to National Tuberculosis Reference Laboratory from the centers that are members of the Tuberculosis Laboratory Surveillance Network. The phylogenetic relationship between 96 MDR-TB strains was investigated with the combination of bead-based spoligotyping and MIRU-VNTR15 methods on the MAGPIX® Milliplex Map device. In this study, it was determined that the T1 family is more common in our country and LAM7-TUR family is less common than the Beijing family unlike other studies. It was determined that the strains in the same cluster had different locus profiles, and there was no transmission from the same clone in the clonal typing we performed with spoligotyping and MIRU-VNTR15.