Multidrug-resistant Tuberculosis Strains Research Articles

2355. Prison as a Driver of Recent Transmissions of Multidrug-Resistant Tuberculosis in Callao, Peru

Abstract Background Peru has one of the highest rates of multidrug-resistant tuberculosis (MDR-TB) in the Latin America region, with Callao being one of the hot spots. We sought to identify resistance patterns and key drivers of recent MDR-TB transmission in Callao, Peru. Methods Cross-sectional study including clinical specimens identified as MDR-TB strains in Callao, Peru between April 2017 and December 2019. DNA was extracted for whole genome sequencing and data used for phylogenetic classification, clustering, and resistance causing mutation analyses. Recent transmission was defined based on an isolate-to-isolate distance of ≤5 (D5) single nucleotide polymorphisms (SNPs). We used logistic regression models to analyze the relationship between MDR-TB clustering and epidemiologic factors including age, sex, history of diabetes mellitus, HIV infection, illicit drug use, known TB contact, prior TB disease, and imprisonment. Results 171 unique MDR-TB strains were included; 93% were assigned to lineage 4 and 7% to lineage 2. The most prevalent sublineage was 4.3.3 LAM (57%), followed by 4.3.4.2 LAM (10%) and 4.1.2.1 Haarlem (9%). In the dominant 4.3.3 LAM sublineage, concomitant resistance was common, including resistance mutations to pyrazinamide (92%), ethambutol (22%), ethionamide (23%), and quinolones (10%); 4 isolates harbored bedaquiline resistance mutations. Seventy-four percent of 4.3.3 LAM isolates were D5-clustered, with 53 (30%) isolates within a single dominant cluster. Male sex (odds ratio [OR], 3.6; 95% confidence interval [CI], 1.5 – 8.9), drug use (OR, 4.9; 95% CI, 1.9 – 12.7), and current or prior history of imprisonment (OR, 12; 95% CI, 3.3 – 43.5) were associated with the dominant D5 cluster in unadjusted analyses. History of imprisonment remained independently associated with the dominant D5 cluster in multivariate analyses (adjusted OR, 8.9; 95% CI, 1.6 – 50.6). History of imprisonment was also independently associated with being part of any D5 clusters in multivariate analyses (adjusted OR, 4.8; 95% CI, 1.2 – 20.3). Conclusion History of imprisonment was linked to current MDR-TB transmissions, indicating an important role of prisons in driving the MDR-TB epidemic. Disclosures Moises A. Huaman, MD, MSc, Gilead: Grant/Research Support|Insmed: Grant/Research Support.

Recognition of specific immunogenic antigens with potential diagnostic value in multi-drug resistant Mycobacterium tuberculosis inducing humoral immunity in MDR-TB patients.

Tuberculosis (TB) as a public health crisis is caused by the intracellular bacterium Mycobacterium tuberculosis. Detection of immunogenic proteins in TB is valuable for the development of diagnostic tests, vaccine formulations and monitoring treatment outcome. In this study, we differentiated the immune-reactivity of proteins in multidrug-resistant tuberculosis (MDRTB) and drug-susceptible strains using purified anti-MDRTB antibodies isolated from inpatients. Our data showed that the anti- MDRTB antibody was well able to detect the MDR strain in the patient's sputum. The immunogenic proteins of MDRTB were purified by affinity chromatography and subjected to matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. Analysis of the data revealed that seven MDRTB immunogenic proteins, including Rv2986c (HupB), Rv3699, Rv1133c (MetE), Rv0440 (GroEL), Rv3057c, Rv2558 and Rv2971 are involved in DNA stability, metabolism, cellular processes and some unknown functions. Similarities in the electrophoresis protein profiles were evident between the extracts of MDR and sensitive TB strains. However, the protein expression patterns of MDRTB isolates were distinguishable from that formed by susceptible TB strains.

MDR-TB in Pakistan: Challenges, efforts, and recommendations

Tuberculosis (TB), a global health concern is also a leading cause of mortality and morbidity across Pakistan affecting a major proportion of the population. The absence of an integrated system to control the spread of TB has led to a rise in multidrug resistant strains of TB (MDR-Tb) which do not exhibit any sensitivity towards the first line therapy for TB. Such adverse circumstances call for effective planning strategies to mitigate the health hazards of MDR-TB. This article briefly highlights the challenges encountered by the already burdened healthcare system and suggests relatively inexpensive approaches to tackle the ongoing crisis associated with MDR-TB on a national scale.

Clinical Features and Outcome of Multidrug-Resistant Osteoarticular Tuberculosis: A 12-Year Case Series from France.

The optimal treatment for osteoarticular infection due to multidrug-resistant tuberculosis strains (MDR-OATB) remains unclear. This study aims to evaluate the diagnosis, management and outcome of MDR-OATB in France. We present a case series of MDR-OATB patients reviewed at the French National Reference Center for Mycobacteria between 2007 and 2018. Medical history and clinical, microbiological, treatment and outcome data were collected. Twenty-three MDR-OATB cases were reported, representing 3% of all concurrent MDR-TB cases in France. Overall, 17 were male, and the median age was 32 years. Six patients were previously treated for TB, including four with first-line drugs. The most frequently affected site was the spine (n = 16). Bone and joint surgery were required in 12 patients. Twenty-one patients (91%) successfully completed the treatment with a regimen containing a mean of four drugs (range, 2–6) for a mean duration of 20 months (range, 13–27). Overall, high rates of treatment success were achieved following WHO MDR-TB treatment guidelines and individualized patient management recommendations by the French National TB Consilium. However, the optimal combination of drugs, duration of treatment and role of surgery in the management of MDR-OATB remains to be determined.

Comprehensive genomic analysis of Mycobacterium tuberculosis reveals limited impact of high-fitness genotypes on MDR-TB transmission

Environmental and host-related factors that contribute to the transmission of multidrug-resistant tuberculosis (MDR-TB) have become an increasing concern, but the impact of bacterial genetic factors associated with bacterial fitness on MDR-TB transmission is poorly understood. Here, we present a global view of the correlation between common fitness-related genotypes and MDR-TB transmission by analyzing a representative number of MDR-TB isolates. We assembled a global whole genome sequencing (WGS) dataset of MDR-TB strains collected through retrospective cohorts or population-based approaches using public databases and literature curation. WGS-based clusters were defined as groups of strains with genomic difference of ≤ 5 SNPs. We curated high-quality WGS data of 4696 MDR-TB isolates from 17 countries with a mean clustering rate of 48% (range 0-100%). Correlational analysis showed that increased risk of MDR-TB strain clustering was not associated with compensatory mutations (OR 1.07, 95% CI 0.72-1.59), low-fitness cost drug-resistant mutations (katG S315T: OR 1.42, 95% CI 0.82-2.47; rpoB S450L: OR 1.26, 95% CI 0.87-1.83) or Lineage 2 (OR 1.50, 95% CI 0.95-2.39). The factors most commonly thought to increase bacterial fitness were not significantly associated with increased MDR-TB transmission, and thus do not appear to be major contributors to the current epidemic of MDR-TB.

World Tuberculosis Day 2022: aligning COVID-19 and tuberculosis innovations to save lives and to end tuberculosis

World Tuberculosis Day 2022: aligning COVID-19 and tuberculosis innovations to save lives and to end tuberculosis

Transmission of multidrug-resistant tuberculosis within family households by DTM-PCR and MIRU-VNTR genotyping

BackgroundDrug-resistant tuberculosis (TB) continues to be a public health threat. There are few studies on transmission and genotyping of MDR-TB family households in China. This study aimed to investigate transmission of multidrug-resistant tuberculosis (MDR-TB) within family households by deletion-targeted multiplex polymerase chain reaction (DTM-PCR), mycobacterial interspersed repetitive unit variable number tandem repeats (MIRU-VNTR) genotyping.MethodsAmong 993 MDR-TB patients registered from Wuhan Institute for Tuberculosis Control, drug resistance and the time interval between the index patients and secondary patients were analyzed in 49 MDR-TB patients from 23 families, in which 22 MDR-TB strains from 11 families who had matched strains were genotyped by DTM-PCR and standard 24-loci MIRU-VNTR genotyping method.ResultsThe time interval between the index patients and the secondary patients ranged from half a month to 110 months. Thirteen secondary patients developed active MDR-TB within two years and accounted for 50% (13/26) of all secondary patients. Among eleven pairs of MDR-TB families, six pairs had identical genotypes, the cluster rate was 54.5% (12/22); three pairs had a single MIRU-VNTR locus variation. If a single MIRU-VNTR locus variation was tolerated in the cluster definition, the cluster rate raised to 81.8% (18/22).ConclusionsThe family households of MDR-TB patients are at risk for infection of MDR-TB. To reduce transmission, MDR-TB patients should be diagnosed earlier and promptly treated in an effective manner, meanwhile, the close family contacts should be screened for TB infection.

Telacebec: an investigational antibacterial for the treatment of tuberculosis (TB)

ABSTRACT Introduction Tuberculosis is an infectious disease that affected more than 50 million people and killed 6.7 million patients in the past 5 years alone. Additionally, rising incidence of treatment resistance threatens the global effort to eradicate this disease. With limited options available, additional novel antibiotics are needed for the treatment of multidrug-resistant tuberculosis (MDR-TB). Telacebec is a first-in-class antibiotic that targets the pathogen’s energy metabolism. Areas covered This paper provides an overview of the recent progress in the development and testing of telacebec. We discuss published clinical data and examine the design and setup of its clinical trials. We also offer insights on the therapeutic potential of telacebec and aspects of which should be evaluated in the future. Expert opinion The first phase 2a trial showed a correlation between dosage and bacterial load in patient sputum, which should be confirmed using a direct measurement method such as colony-forming unit counting. Its clinical efficacy, favorable pharmacokinetic properties, low arrhythmogenic risk, and activity against MDR-TB strains make telacebec a suitable candidate for further development. Future clinical testing in combination with approved second-line drugs will reveal its full potential against MDR-TB. Considering recent preclinical studies, we also recommend initiating clinical trials for Buruli ulcer and leprosy.

Application of Lipid-Based Nanocarriers for Antitubercular Drug Delivery: A Review.

The antimicrobial drugs currently used for the management of tuberculosis (TB) exhibit poor bioavailability that necessitates prolonged treatment regimens and high dosing frequency to achieve optimal therapeutic outcomes. In addition, these agents cause severe adverse effects, as well as having detrimental interactions with other drugs used in the treatment of comorbid conditions such as HIV/AIDS. The challenges associated with the current TB regimens contribute to low levels of patient adherence and, consequently, the development of multidrug-resistant TB strains. This has led to the urgent need to develop newer drug delivery systems to improve the treatment of TB. Targeted drug delivery systems provide higher drug concentrations at the infection site, thus leading to reduced incidences of adverse effects. Lipid-based nanocarriers have proven to be effective in improving the solubility and bioavailability of antimicrobials whilst decreasing the incidence of adverse effects through targeted delivery. The potential application of lipid-based carriers such as liposomes, niosomes, solid lipid nanoparticles, nanostructured lipid carriers, nano and microemulsions, and self-emulsifying drug delivery systems for the treatment of TB is reviewed herein. The composition of the investigated lipid-based carriers, their characteristics, and their influence on bioavailability, toxicity, and sustained drug delivery are also discussed. Overall, lipid-based systems have shown great promise in anti-TB drug delivery applications. The summary of the reviewed data encourages future efforts to boost the translational development of lipid-based nanocarriers to improve TB therapy.

Antimycobacterial activity of acetone extract and isolated metabolites from folklore medicinal lichen Usnea laevis Nyl. against drug-sensitive and multidrug-resistant tuberculosis strains

Ethnopharmacological relevanceTuberculosis (Tb) is one of the most infectious diseases caused by Mycobacterium tuberculosis (M.t) with almost 2 million deaths yearly. Although many Tb control programs have been organised, there is an elevated number of Tb cases due to the appearance of extremely drug-resistant and multidrug-resistant (MDR) Tb strains. In the cultures of Venezuelan Andes, fruticose lichen Usnea laevis Nyl. (Usneaceae) with folklore name ‘Barba de Piedra, Tusinya’ is used as a natural remedy for Tb. Aim of the studyThis study was performed to provide a scientific rationale for the folklore usage of U. laevis in treating Tb by validating its antimycobacterial activity against two drug-sensitive and four MDR-Tb strains. Materials and methodsThe mycobacterial inhibitory activities of acetone extract (Ul), fractions (F1-10), and isolated metabolites (1–4) of U. laevis were evaluated against M.t H37Ra using 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide reduction menadione assay (XRMA). Furthermore, Ul and 1–4 were subjected to antimycobacterial activity against M.t H37Ra, Mycobacterium smegmatis, and four MDR-Tb (MDR-A8, MDR-V791, MDR-R and MDR-40) strains using resazurin microtitre plate assay (REMA) and cytotoxicity against THP-1 macrophages using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and their selectivity index values were also calculated. ResultsInitially, Ul has shown prominent inhibitory activity (IC50 value: 5.44 ± 0.36 μg/ml) and four of its fractions (F1, F2, F5 and F7) also exhibited the best inhibitory activity (IC50 values ranged from 7.46 ± 0.19 to 71.38 ± 2.57 μg/ml) against M.t H37Ra using XRMA. Purification of these bioactive fractions identified four metabolites, namely usnic acid (1), atranorin (2), salazinic acid (3), and lobaric acid (4). From the MIC values of REMA, it was identified that Ul, 1 and 4 were more effective in inhibiting the growth of all four MDR-Tb strains, compared to first-line drug rifampicin. Interestingly, Ul has shown better antimycobacterial activity than 1–4 and rifampicin against MDR-Tb strains may be due to the synergistic effect of its metabolites. Also, the IC50 values of Ul and 1–4 on THP-1 macrophages were found to be far higher than MIC values against tested Tb strains, indicating that THP-1 macrophages were not harmfully affected at concentrations that were effective against Tb strains. Further, the calculated selectivity index values revealed the more active and non-toxicity of Ul, 1 and 4 against MDR-Tb strains than rifampicin. ConclusionsThe current study lends the first evidence for the presence of antimycobacterial metabolites in U. laevis. The results exposed the Andean folklore use of U. laevis for treating Tb, and the key biomarker metabolites were found to be 1 and 4. Hence, it can be concluded that U. laevis can be used as a potential source for the novel drug development for MDR-Tb.

Genetic Diversity and Drug Susceptibility Profiles of Multidrug-Resistant Tuberculosis Strains in Southeast China.

BackgroundMultidrug-resistant tuberculosis (MDR-TB) isolates collected from Fujian province, China were assessed for molecular epidemiological characteristics. Analysis of isolate genotype profiles revealed that the Beijing genotype was associated with especially high drug resistance and community transmission rates.MethodsA total of 119 MDR-TB isolates obtained from TB patients in Fujian province were typed using 24–locus mycobacterium interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing and spoligotyping. Drug susceptibility testing of all isolates was conducted using the L-J proportion method, with pyrazinamide (PZA) susceptibility testing conducted using the Mycobacterium Growth Indicator Tube System 960 (MGIT 960).ResultsWe obtained 26 spoligotypes for the 119 isolates examined in this work. Spoligotyping results revealed that 80 (67.2%) isolates possessed the Beijing family genotypic profiles. Patients aged 25–44 years and ≥45 years were most likely to be infected by non-Beijing genotypes. The percentage of clustered cases with both PZA and ofloxacin (OFLX) resistance was significantly greater than the corresponding percentage for non-clustered cases. Of 44 PZA-resistant isolates, 28 isolates (63.6%) harbored pncA mutations, while pncA mutations were only detected in 7 (9.3%) PZA-susceptible isolates.ConclusionOur data demonstrate that the Beijing genotype is the dominant lineage among MDR-TB strains circulating in Fujian. Thus, MDR-TB infections occurring within this province are not likely associated with recent transmission events. PZA and fluoroquinolone resistance profiles were found to be associated with clustered isolates. Mutation of pncA is the main driver of MDR-TB PZA resistance and is associated with mutation sites scattered throughout the entire pncA protein-coding region.

English

The control of tuberculosis (TB) has become a global health challenge due to the emergence of multidrug-resistant tuberculosis (MDR-TB) in Mycobacterium tuberculosis (MTB). This highlights the need for faster and more accurate detection of tuberculosis cases. The study aims to detect MDR-TB strains of pulmonary tuberculosis using resistance ratio method and to compare the diagnostic value of drug susceptibility testing (DST) with line probe assay (LPA) using Genotype MTBDRplus. All the sputum samples were tested for Acid Fast Bacilli (AFB) by Ziehl-Neelsen’s staining method and were cultured on Lowenstein-Jensen (L-J) media. The identification and confirmation of M. tuberculosis were done using various biochemical tests. DST was carried against the first line of anti-TB drugs. MTB positive samples were subjected to LPA. A total of 57 samples were subjected to DST and LPA for the detection of drug resistance of MTB to RIF and INH after conventional detection methods were applied to all the samples. Among these 57 MTB samples, 11 (19.29%) were resistant to INH, 4 (7.14%) were resistant to RIF; eleven (19.29%) isolates were identified as MDR-TB. LPA revealed 54 MTB positive among 57 MTB culture-positive samples and 3 showed invalid results. In LPA, MDR-TB was found in10 samples (17.54%) in which one was RIF-resistant. The study concludes risk factors that resulted in the development of TB are biomedical, socio-cultural, and behavioral interactions. LPA can be used as a rapid diagnostic technique for the detection of MDR-TB. Key words: Tuberculosis (TB), multidrug resistance (MDR-TB), drug susceptibility testing (DST), line probe assay (LPA). &nbsp

Genomic epidemiology of tuberculosis in eastern Malaysia: insights for strengthening public health responses.

Tuberculosis is a leading public health priority in eastern Malaysia. Knowledge of the genomic epidemiology of tuberculosis can help tailor public health interventions. Our aims were to determine tuberculosis genomic epidemiology and characterize resistance mutations in the ethnically diverse city of Kota Kinabalu, Sabah, located at the nexus of Malaysia, Indonesia, Philippines and Brunei. We used an archive of prospectively collected Mycobacterium tuberculosis samples paired with epidemiological data. We collected sputum and demographic data from consecutive consenting outpatients with pulmonary tuberculosis at the largest tuberculosis clinic from 2012 to 2014, and selected samples from tuberculosis inpatients from the tertiary referral centre during 2012–2014 and 2016–2017. Two hundred and eight M . tuberculosis sequences were available for analysis, representing 8 % of cases notified during the study periods. Whole-genome phylogenetic analysis demonstrated that most strains were lineage 1 (195/208, 93.8 %), with the remainder being lineages 2 (8/208, 3.8 %) or 4 (5/208, 2.4 %). Lineages or sub-lineages were not associated with patient ethnicity. The lineage 1 strains were diverse, with sub-lineage 1.2.1 being dominant (192, 98 %). Lineage 1.2.1.3 isolates were geographically most widely distributed. The greatest diversity occurred in a border town sub-district. The time to the most recent common ancestor for the three major lineage 1.2.1 clades was estimated to be the year 1966 (95 % HPD 1948–1976). An association was found between failure of culture conversion by week 8 of treatment and infection with lineage 2 (4/6, 67 %) compared with lineage 1 strains (4/83, 5 %) (P<0.001), supporting evidence of greater virulence of lineage 2 strains. Eleven potential transmission clusters (SNP difference ≤12) were identified; at least five included people living in different sub-districts. Some linked cases spanned the whole 4-year study period. One cluster involved a multidrug-resistant tuberculosis strain matching a drug-susceptible strain from 3 years earlier. Drug resistance mutations were uncommon, but revealed one phenotype–genotype mismatch in a genotypically multidrug-resistant isolate, and rare nonsense mutations within the katG gene in two isolates. Consistent with the regionally mobile population, M. tuberculosis strains in Kota Kinabalu were diverse, although several lineage 1 strains dominated and were locally well established. Transmission clusters – uncommonly identified, likely attributable to incomplete sampling – showed clustering occurring across the community, not confined to households or sub-districts. The findings indicate that public health priorities should include active case finding and early institution of tuberculosis management in mobile populations, while there is a need to upscale effective contact investigation beyond households to include other contacts within social networks.

Clofazimine functionalized polymeric nanoparticles for brain delivery in the tuberculosis treatment

Central nervous system tuberculosis (CNS-TB) is the most severe form of the disease especially due to the inability of therapeutics to cross the blood–brain barrier (BBB). Clofazimine (CFZ) stands out for presenting high in vitro activity against multi-drug resistant strains of Mycobacterium tuberculosis, however, CFZ physicochemical and pharmacokinetics properties limit drug penetration into the CNS and, consequently, its clinical use. The aim of this work was to develop polymeric nanoparticles (NPs) of poly(lactic-co-glycolic acid) (PLGA) and polyethylene glycol (PEG) loaded with CFZ and functionalized with a transferrin receptor (TfR)-binding peptide, aiming brain drug delivery for CNS-TB treatment by the intravenous route. The poor water solubility and high lipophilicity of CFZ was overcome through its entrapment into PLGA-PEG NPs manufactured by both conventional and microfluidic techniques using the nanoprecipitation principle. In vitro studies in brain endothelial hCMEC/D3 cells demonstrated that CFZ incorporation into the NPs was advantageous to reduce drug cytotoxicity. The TfR-binding peptide-functionalized NPs showed superior cell interaction and higher CFZ permeability across hCMEC/D3 cell monolayers compared to the non-functionalized NP control, thus indicating the efficacy of the functionalization strategy on providing CFZ transport through the BBB in vitro. The functionalized NPs demonstrate suitability for CFZ biological administration, suggested with low plasma protein binding, off-target biodistribution and precise delivery of CFZ towards the brain parenchyma.

Ethionamide in MDR TB: The fall of a legend

The prime focus of this small review article is about an anti-tubercular drug which in spite of being a well-known and good second line bactericidal drug against multi-drug resistant (MDR) strains of Mycobacterium Tuberculosis (MTB) has been relegated amongst the last but second of the Group C drugs in the current WHO-recommended Classification of drugs for designing a Multi-Drug Resistant (MDR) TB regimen. We now discuss this startling fall of a legendary anti-tubercular drug in the arsenal against fighting the scourge of MDR TB.

Community transmission of multidrug-resistant tuberculosis is associated with activity space overlap in Lima, Peru

BackgroundTransmission of multidrug-resistant tuberculosis (MDRTB) requires spatial proximity between infectious cases and susceptible persons. We assess activity space overlap among MDRTB cases and community controls to identify potential areas of transmission.MethodsWe enrolled 35 MDRTB cases and 64 TB-free community controls in Lima, Peru. Cases were whole genome sequenced and strain clustering was used as a proxy for transmission. GPS data were gathered from participants over seven days. Kernel density estimation methods were used to construct activity spaces from GPS locations and the utilization distribution overlap index (UDOI) was used to quantify activity space overlap.ResultsActivity spaces of controls (median = 35.6 km2, IQR = 25.1–54) were larger than cases (median = 21.3 km2, IQR = 17.9–48.6) (P = 0.02). Activity space overlap was greatest among genetically clustered cases (mean UDOI = 0.63, sd = 0.67) and lowest between cases and controls (mean UDOI = 0.13, sd = 0.28). UDOI was positively associated with genetic similarity of MDRTB strains between case pairs (P < 0.001). The odds of two cases being genetically clustered increased by 22% per 0.10 increase in UDOI (OR = 1.22, CI = 1.09–1.36, P < 0.001).ConclusionsActivity space overlap is associated with MDRTB clustering. MDRTB transmission may be occurring in small, overlapping activity spaces in community settings. GPS studies may be useful in identifying new areas of MDRTB transmission.

82125 Multiple epidemics of multidrug-resistant tuberculosis revealed by spatial disease mapping and whole-genome sequencing analysis in urban China

ABSTRACT IMPACT: Our study will integrate state-of-the-art methods in pathogen genomics, epidemiology, and geospatial analysis to identify both host- and pathogen-factors driving the MDR-TB transmission and the study outcome can inform the design of targeted interventions OBJECTIVES/GOALS: The emergence of multidrug-resistant tuberculosis (MDR-TB) poses serious challenges for the global eradication of tuberculosis. Recent research has shown that transmission is now the dominant driver of MDR-TB. However, our limited understanding of where and among whom MDR-TB is transmitted hampers efforts to control person-to-person spread. METHODS/STUDY POPULATION: We used several analytic approaches to characterize the dynamics of MDR-TB transmission in Shanghai, China. We identified all culture-confirmed MDR cases between 2009-2016 in the city and 1) estimated individual-level risk factors for MDR disease; 2) mapped the TB cases by their home addresses and used a Bayesian spatial disease mapping method to identify regions with an elevated risk of MDR-TB; and 3) we sequenced all MDR isolates to understand whether transmission explained variance in risk that was not attributable to the distribution of individual or location-specific risk variates. RESULTS/ANTICIPATED RESULTS: There were 1034 MDR-TB cases among 16,315 culture-confirmed TB cases during the study period. Bayesian disease mapping identified spatial heterogeneity of MDR-TB and determined four hotspots with an elevated risk of MDR-TB, none of which were fully explained by individual or regional-covariates (Figure 1). Sequencing revealed that more than 40% of the MDR-TB strains were in genomic clusters, indicating recent MDR-TB transmission. Most importantly, MDR-TB cases in three of the four large clades (&gt;8 isolates) were spatially concentrated in three strain-specific hotspots (Figure 2). DISCUSSION/SIGNIFICANCE OF FINDINGS: With the combination of traditional epidemiological tools, geographical, and genomic methods, this study revealed multiple loci of transmission of specific MDR-TB clades within a single city. Identification of where and among whom MDR-TB is transmitted can inform the design of targeted interventions.

Fidaxomicin has high in vitro activity against Mycobacterium tuberculosis.

This study aimed to evaluate whether the antibiotic fidaxomicin has in vitro activity against Mycobacterium tuberculosis (Mtb). 38 fully drug-sensitive Mtb strains and 34 multidrug-resistant tuberculosis (MDR-TB) strains were tested using the microplate alamar blue assay (MABA) method to determine the minimum inhibitory concentrations (MICs) for fidaxomicin and rifampicin. Fidaxomicin has high in vitro activity against Mtb and is a potential drug to treat Mtb, and MDR-TB infections in particular.

Prevalence and molecular characterizations of seven additional drug resistance among multidrug-resistant tuberculosis in China: A subsequent study of a national survey

The drug resistance prevalence data facilitates selection of the initial drug for treating multidrug-resistant tuberculosis (MDR-TB). The aim of this study was to investigate the prevalence and molecular characterization of seven additional types of drug resistances among MDR-TB isolates collected from the first/only nationwide drug resistance surveillance in China. A total of 391 out of the 401 MDR-TB strains were successfully recovered by Löwenstein-Jensen medium. Drug susceptibility testing was performed against moxifloxacin (Mfx), bedaquiline (Bdq), linezolid (Lzd), clofazimine (Cfz), cycloserine (Cs), delamanid (Dlm) and pyrazinamide (PZA). The strains were subjected to whole-genome sequencing for the analysis corresponding drug resistant genes and their profiles. 269 (68.80%) were simple MDR-TB, 28 (7.16%) were extensively drug-resistant tuberculosis (XDR-TB) and 94 (24.04%) were pre-XDR-TB. Dlm, Lzd, Cfz and Bdq presented the lowest drug resistant rates i.e. 3.32% (13/391), 3.84% (15/391),6.65% (26/391) and 7.16% (28/391), respectively. Mfx (17.39%, 68/391) and CS (13.55%, 53/391) also demonstrated strong potencies against the MDR strains, whereas PZA (38.36%, 150/391) presented much higher resistant rate. 54.41% (37/68) Mfx-resistant strains carried mutations located within gyrA or gyrB. 70.15% (94/134) PZA-resistant strains had pncA mutations. Two of the 26 Cfz-resistant isolates had mutation in Rv0678 were also resistant to Bdq. Dlm, Lzd, Cfz and Bdq exhibited excellent activity against MDR-TB, including XDR-TB. These data highlighted the necessity of a timely, feasible and reliable DST, while genotypic DST for Mfx and PZA is promising at this moment.

Comparative evaluation of nitrate reductase assay with conventional proportion method for isoniazid and rifampicin susceptibility testing in smear positive pulmonary tuberculosis patients

With the surge of multidrug-resistant tuberculosis (MDR-TB) strains there is an escalating need for precise and cost-effective methods for a precipitous diagnostic and drug susceptibility testing (DST), chiefly in resource limited countries where tuberculosis is endemic. To find out the susceptibility pattern of Isoniazid (INH) and Rifampicin (RMP) in smear positive cases of pulmonary TB at MMIMSR Mullana and to evaluate the sensitivity and specificity of nitrate reductase assay (NRA) with standard proportion method for INH and RMP susceptibility testing. A cross sectional test validation study was carried out at Mycobacteriology division in the Department of Microbiology on 100 smear positive sputum samples from pulmonary tuberculosis patients.In comparison with L.J proportion method, NRA showed sensitivity, specificity, PPV, NPV, accuracy of 100%, 98.64%, 87.50%, 100%, and 98.76% respectively for INH. The sensitivity, specificity, PPV, NPV, accuracy of NRA compared to PM for RMP was 100%, 97.33%, 75.00%, 100%, 97.53% respectively. Direct NRA is simple, easy to perform, prompt, reasonably less expensive, without requisite of expensive reagents and advanced instrumentation. It is highly specific and sensitive technique in the detection of drug resistant TB when tested in parallel with the proportion method. Therefore, NRA may be routinely employed in TB laboratories in developing countries for drug susceptibility testing of .