Multidrug-resistant Cases Research Articles

Whole-genome sequencing of extensively drug-resistant Salmonella enterica serovar Typhi clinical isolates from the Peshawar region of Pakistan

BackgroundTyphoid fever, caused by Salmonella enterica serovar Typhi, is a significant public health concern due to the escalating of antimicrobial resistance (AMR), with limited treatment options for extensively drug-resistant (XDR) S. Typhi strains pose a serious threat to disease management and control. This study aimed to investigate the genomic characteristics, epidemiology and AMR genes of XDR S. Typhi strains from typhoid fever patients in Pakistan. MethodologyWe assessed 200 patients with enteric fever symptoms, confirming 65 S. Typhi cases through culturing and biochemical tests. Subsequent antimicrobial susceptibility testing revealed 40 cases of extensively drug-resistant (XDR) and 25 cases of multi-drug resistance (MDR). Thirteen XDR strains were selected for whole-genome sequencing, to analyze their sequence type, phylogenetics, resistance genes, pathogenicity islands, and plasmid sequences using variety of data analysis resources. Pangenome analysis was conducted for 140 XDR strains, including thirteen in-house and 127 strains reported from other regions of Pakistan, to assess their genetic diversity and functional annotation. ResultsMLST analysis classified all isolates as sequence type 1 (ST-1) with 4.3.1.1. P1 genotype characterization. Prophage and Salmonella Pathogenicity Island (SPI) analysis identified intact prophages and eight SPIs involved in Salmonella's invasion and replication within host cells. Genome data analysis revealed numerous AMR genes including dfrA7, sul1, qnrS1, TEM-1, Cat1, and CTX-M-15, and SNPs associated with antibiotics resistance. IncY, IncQ1, pMAC, and pAbTS2 plasmids, conferring antimicrobial resistance, were detected in a few XDR S. Typhi strains. Phylogenetic analysis inferred a close epidemiological linkage among XDR strains from different regions of Pakistan. Pangenome was noted closed among these strains and functional annotation highlighted genes related to metabolism and pathogenesis. ConclusionThis study revealed a uniform genotypic background among XDR S. Typhi strains in Pakistan, signifying a persistence transmission of a single, highly antibiotic-resistant clone. The closed pan-genome observed underscores limited genetic diversity and highlights the importance of genomic surveillance for combating drug-resistant typhoid infections.

Role of the first WHO mutation catalogue in the diagnosis of antibiotic resistance in Mycobacterium tuberculosis in the Valencia Region, Spain: a retrospective genomic analysis

In June, 2021, WHO published the most complete catalogue to date of resistance-conferring mutations in Mycobacterium tuberculosis. Here, we aimed to assess the performance of genome-based antimicrobial resistance prediction using the catalogue and its potential for improving diagnostics in a real low-burden setting. In this retrospective population-based genomic study M tuberculosis isolates were collected from 25 clinical laboratories in the low-burden setting of the Valencia Region, Spain. Culture-positive tuberculosis cases reported by regional public health authorities between Jan 1, 2014, and Dec 31, 2016, were included. The drug resistance profiles of these isolates were predicted by the genomic identification, via whole-genome sequencing (WGS), of the high-confidence resistance-causing variants included in the catalogue and compared with the phenotype. We determined the minimum inhibitory concentration (MIC) of the isolates with discordant resistance profiles using the resazurin microtitre assay. WGS was performed on 785 M tuberculosis complex culture-positive isolates, and the WGS resistance prediction sensitivities were: 85·4% (95% CI 70·8-94·4) for isoniazid, 73·3% (44·9-92·2) for rifampicin, 50·0% (21·1-78·9) for ethambutol, and 57·1% (34·0-78·2) for pyrazinamide; all specificities were more than 99·6%. Sensitivity values were lower than previously reported, but the overall pan-susceptibility accuracy was 96·4%. Genotypic analysis revealed that four phenotypically susceptible isolates carried mutations (rpoB Leu430Pro and rpoB Ile491Phe for rifampicin and fabG1 Leu203Leu for isoniazid) known to give borderline resistance in standard phenotypic tests. Additionally, we identified three putative resistance-associated mutations (inhA Ser94Ala, katG Leu48Pro, and katG Gly273Arg for isoniazid) in samples with substantially higher MICs than those of susceptible isolates. Combining both genomic and phenotypic data, in accordance with the WHO diagnostic guidelines, we could detect two new multidrug-resistant cases. Additionally, we detected 11 (1·6%) of 706 isolates to be monoresistant to fluoroquinolone, which had been previously undetected. We showed that the WHO catalogue enables the detection of resistant cases missed in phenotypic testing in a low-burden region, thus allowing for better patient-tailored treatment. We also identified mutations not included in the catalogue, relevant at the local level. Evidence from this study, together with future updates of the catalogue, will probably lead in the future to the partial replacement of culture testing with WGS-based drug susceptibility testing in our setting. European Research Council and the Spanish Ministerio de Ciencia.

Efficacy of Antibiotic Bone Cement in the Treatment of Burkholderia cepacia Infection After Spinal Internal Fixation Surgery: Case Report and Literature Review

In recent years, the number of spinal internal fixation operations has increased significantly, correlating with an elevated risk of postoperative surgical site infection and a rising incidence rate. While the conventional treatment approach involves surgical debridement combined with antibiotic administration, there is a notable gap in reported strategies for Burkholderia cepacia infection and patients exhibiting multidrug resistance. Surgical site infection occurred in a patient following internal fixation surgery for thoracic vertebral fractures. Despite the application of systemic antibiotics and regular dressing changes, no improvement was observed. Bacterial culture and drug sensitivity experiments revealed a multidrug-resistant Burkholderia cepacia infection. Two comprehensive debridement procedures were performed along with continuous post-operative irrigation combined with antibiotic administration; however, no significant improvement was observed. The patient's infection was significantly controlled following treatment with vancomycin loaded bone cement. Following spinal internal fixation surgery, the management of a B.cepacian infection with multidrug resistance presented a significant challenge, despite the application of debridement procedures and systemic antibiotics. In this case, after 20days of treatment with vancomycin-loaded bone cement, the patient's C-reactive protein level decreased to 54mg/L, was normalized by February, and normal levels were maintained in the surgical area 1month and 6months after bone cement removal. The use of vancomycin-loaded bone cement proves effective in treating postoperative B.cepacian infection in a multidrug-resistant case following spinal internal fixation surgery.

Multidrug Resistance among Patients with Surgical Site Infection in a Tertiary Care Centre.

Infections at surgical sites are a major problem all over the world. Pathogens linked to postoperative infections are becoming increasingly resistant to antibiotics, which presents a significant therapeutic challenge for surgeons and raises the financial burden placed on patients. The study aimed to find the prevalence of multidrug resistance among patients with surgical site infections in a tertiary care centre. A descriptive cross-sectional study was conducted from 3 November 2022 to 2 May 2023 among post-operative patients with surgical site infection after receiving ethical approval from the Institutional Review Committee. Swab samples were sent for antimicrobial susceptibility testing. Convenience sampling method was used. The point estimate was calculated at a 95% Confidence Interval. Among 147 patients with surgical site infection, the prevalence of multi-drug resistance was 95 (64.63%) (56.90-72.36, 95% Confidence Interval). Among them, 51 (53.68%) patients were in the 21-40 years age group and most of them were male 53 (55.79%). The prevalence of multidrug-resistant cases was similar to other studies done in similar settings. antimicrobial drug resistance; multidrug resistance; Staphylococcus aureus; surgical site infection.

Validation of public stigma of tuberculosis scale during the COVID-19 pandemic using the Rasch model.

Tuberculosis (TB) is still a major global health problem, and it has been particularly concerning during the COVID-19 pandemic. Non-compliance with anti-TB treatment increases the number of multidrug-resistant cases, causing ongoing transmission and increased morbidity and mortality. The main factors causing TB patients' non-compliance are stigma and lack of financial resources. Stigma harms patients and may cause them to delay seeking and adhering to treatment. Thus, it is important to measure the public stigma surrounding TB. However, few scales are available to measure this stigma as it developed during the COVID-19 pandemic. This study, therefore, aimed to develop and validate such a scale. Mixed methods were employed in this study, consisting of a qualitative phase using in-depth interviews with 26 community leaders and a descriptive quantitative survey of 37 people in the Sumedang District to validate the public stigma of tuberculosis scale during the COVID-19 pandemic. The qualitative data were analyzed using thematic analysis, and the quantitative data were analyzed using the Rasch model. The 21 items yielded by an initial qualitative analysis of the data gathered were validated using the RASCH model, yielding 17 valid items with a Cronbach's Alpha of 0.95, person separation of 3.61, real root mean square deviation (RMSE) of 0.37, infit mean square (INFIT MNSQ) of > +1.25, differential item functioning (DIF) of 1.000, the raw variance of 52.4%, and an unexplained variance ranging from 3.4% to 6.9%. The scale developed to measure the public stigma surrounding TB during the COVID-19 pandemic is valid and reliable to measure stigma surrounding TB in the community, especially the pandemic. Further research is needed to apply the scale to bigger and broader populations to evaluate its measurement consistency.

Discovery, Synthesis, and Optimization of 1,2,4-Triazolyl Pyridines Targeting Mycobacterium tuberculosis.

The rise in multidrug resistant tuberculosis cases underscores the urgent need to develop new treatment strategies for tuberculosis. Herein, we report the discovery and synthesis of a new series of compounds containing a 3-thio-1,2,4-triazole moiety that show inhibition of Mycobacterium tuberculosis (Mtb) growth and survival. Structure-activity relationship studies led us to identify several potent analogs displaying low micromolar to nanomolar inhibitory activity, specifically against Mtb. The potent analogs demonstrated no cytotoxicity in mammalian cells at over 100 times the effective concentration required in Mtb and were bactericidal against Mtb during infection of macrophages. In the exploratory ADME investigations, we observed suboptimal ADME characteristics, which prompted us to identify potential metabolic liabilities for further optimization. Our preliminary investigations into the mechanism of action suggest that this series is not engaging the promiscuous targets that arise from many phenotypic screens. We selected for resistant mutants with the nanomolar potent nitro-containing compound 20 and identified resistant isolates with mutations in genes required for coenzyme F420 biosynthesis and the nitroreductase Ddn. This suggests that the aromatic nitro-1,2,4-triazolyl pyridines are activated by F420-dependent Ddn activity, similar to the nitro-containing TB drug pretomanid. We were able to circumvent the requirement for F420-dependent Ddn activity using compounds that contained non-nitro groups, identifying a key feature to be modified to avoid this predominant resistance mechanism. These studies provide the foundation for the development of a new class of 1,2,4-triazole compounds for the treatment of tuberculosis.

Predictive capabilities of baseline radiological findings for early and late disease outcomes within sensitive and multi-drug resistant tuberculosis cases

PurposeThis study compares performance of Timika Score to standardized, detailed radiologist observations of Chest X rays (CXR) for predicting early infectiousness and subsequent treatment outcome in drug sensitive (DS) or multi-drug resistant (MDR) tuberculosis cases. It seeks improvement in prediction of these clinical events through these additional observations. MethodThis is a retrospective study analyzing cases from the NIH/NIAID supported TB Portals database, a large, trans-national, multi-site cohort of primarily drug-resistant tuberculosis patients. We analyzed patient records with sputum microscopy readings, radiologist annotated CXR, and treatment outcome including a matching step on important covariates of age, gender, HIV status, case definition, Body Mass Index (BMI), smoking, drug use, and Timika Score across resistance type for comparison. Results2142 patients with tuberculosis infection (374 with poor outcome and 1768 with good treatment outcome) were retrospectively reviewed. Bayesian ANOVA demonstrates radiologist observations did not show greater predictive ability for baseline infectiousness (0.77 and 0.74 probability in DS and MDR respectively); however, the observations provided superior prediction of treatment outcome (0.84 and 0.63 probability in DS and MDR respectively). Estimated lung abnormal area and cavity were identified as important predictors underlying the Timika Score’s performance. ConclusionsTimika Score simplifies the usage of baseline CXR for prediction of early infectiousness of the case and shows comparable performance to using detailed, standardized radiologist observations. The score’s utility diminishes for treatment outcome prediction and is exceeded by the usage of the detailed observations although prediction performance on treatment outcome decreases especially in MDR TB cases.

Pulmonary tuberculosis case notification and burden of drug resistance among children under 15 years of age in Ethiopia: sub-analysis from third-round drug resistance tuberculosis survey

IntroductionData on the burden of bacteriologically confirmed childhood Tuberculosis (PTB) and drug-resistant TB in Ethiopia is limited due to difficulties related to its diagnosis in this population. Therefore, this study aimed to assess bacteriologically confirmed childhood PTB Case Notification Rates (CNRs) and the burden of Drug Resistant-Tuberculosis among children in Ethiopia.MethodRetrospective secondary clinical and laboratory data were obtained from 3rd round national DR-TB survey which was conducted between August 2017 and January 2019. We used IBM SPSS 24 for sub-analysis of 3rd round Drug Resistant-Tuberculosis data. Descriptive statistics were used in computing the association between the sociodemographic characteristics and PTB CNRs, and the strength of the associations was determined using binary logistic regression with Odds ratios (OR) with a 95% confidence interval (CI).ResultOverall, 102 bacteriologically confirmed childhood PTB cases were identified with a median age of 12 (range 1–14) years. Of these, 54 (52.9%) were females and 81 (79.4%) lived in rural areas. HIV-TB co-infection cases were 5/102 (4.3%) and the majority (98%) of cases were newly diagnosed children. Nationally, the incidence of bacteriologically confirmed childhood PTB was calculated to be 5.1 per 100,000 children. The burden of Drug Resistant-Tuberculosis to at least one of the five first-line anti-TB drugs tested was five (6.5%) cases and one (1.3%) was found to be a Multi-drug resistant tuberculosis case. Drug-resistant tuberculosis was significantly associated with the age group 10–14 years (P = 0.002; [AOR] 29.76; [95% CI, 3.51-252.64]) and children living in urban areas (P = 0.027; [AOR] 5.76; 95% CI, 1.22–27.09).ConclusionBacteriologically confirmed childhood PTB cases increased as the age of the children increased. Most of the bacteriologically confirmed childhood PTB and the identified drug Resistant-Tuberculosis cases were new cases. Also, rural children were more affected by TB than their urban, counterparts Drug Resistant-Tuberculosis was higher in urban resident children.

Macrophage targeted polymeric curcumin nanoparticles limit intracellular survival of Mycobacterium tuberculosis through induction of autophagy and augment anti-TB activity of isoniazid in RAW 264.7 macrophages.

Rapid emergence of antibiotic resistance in tuberculosis has left us with limited resources to treat and manage multi drug resistant (MDR) cases of tuberculosis, prompting the development of novel therapeutics. Mycobacterium tuberculosis (MTB) perturbs the host protective pathways for its survival, therefore host directed therapeutic (HDT) interventions offer an attractive alternative strategy. Curcumin (CMN), the principle curcuminoid from Curcuma longa is known to have anti-TB activity against MDR strains of MTB in macrophages. We discovered that treatment of CMN induced autophagy in uninfected and MTB infected macrophages which was evident by conversion of LC3-I to LC3-II and degradation of p62. Inhibition of autophagy by a pharmacological inhibitor 3-MA resulted in significant inhibition of intracellular killing activity of CMN, suggesting the involvement of autophagy in intracellular clearance of MTB. Moreover, annexin v-FITC/PI staining data suggested induction of apoptosis in uninfected and MTB infected macrophages post CMN treatment. This finding was further corroborated by up-regulated expression of pro-apoptotic proteins, Bax, cleaved caspase-3 and PARP and diminished expression of anti-apoptotic protein Bcl-2 as evaluated by immunoblotting. Using GFP-MTB H37Rv and Lysotracker Red staining we demonstrated co-localization of GFP-MTB H37Rv containing phagosome to lysosome after CMN treatment, indicating enhanced phagosome lysosome fusion. Due to poor bioavailability of CMN, its clinical use is limited, therefore to overcome this issue, CMN was encapsulated in Poly(lactic-co-glycolic) acid (PLGA) shell, resulting in polymeric CMN nano particles (ISCurNP). Flow cytometric evaluation suggested >99% uptake of ISCurNP after 3h of treatment. In BALB/c mice, oral dose of ISCurNP resulted in 6.7-fold increase in the bioavailability compared to free CMN. Moreover, ISCurNP treatment resulted in significant decrease in the intracellular survival of MTB H37Rv through induction of autophagy. Adjunct action of ISCurNP and CMN in combination with isoniazid (INH) revealed >99% decrease in intracellular survival of MTB in macrophage as compared to ISCurNP, CMN or INH alone. In conclusion, our findings suggest the role of ISCurNP as novel host directed formulation to combat both sensitive and MDR strains of MTB by induction of autophagy.

Machine Learning of the Whole Genome Sequence of Mycobacterium tuberculosis: A Scoping PRISMA-Based Review.

Tuberculosis (TB) remains one of the most significant global health problems, posing a significant challenge to public health systems worldwide. However, diagnosing drug-resistant tuberculosis (DR-TB) has become increasingly challenging due to the rising number of multidrug-resistant (MDR-TB) cases, despite the development of new TB diagnostic tools. Even the World Health Organization-recommended methods such as Xpert MTB/XDR or Truenat are unable to detect all the Mycobacterium tuberculosis genome mutations associated with drug resistance. While Whole Genome Sequencing offers a more precise DR profile, the lack of user-friendly bioinformatics analysis applications hinders its widespread use. This review focuses on exploring various artificial intelligence models for predicting DR-TB profiles, analyzing relevant English-language articles using the PRISMA methodology through the Covidence platform. Our findings indicate that an Artificial Neural Network is the most commonly employed method, with non-statistical dimensionality reduction techniques preferred over traditional statistical approaches such as Principal Component Analysis or t-distributed Stochastic Neighbor Embedding.

Clinical, Bacteriological Profile and Outcome of Neonatal Sepsis.

Sepsis is a major cause of morbidity and mortality among neonates in Nepal. This study was conducted to determine the clinical-bacteriological profile, their antibiotic susceptibility patterns, and clinical outcome of culture-positive neonatal sepsis. This was a prospective study conducted at B.P Koirala Institute of Health Sciences from July 2018 to June 2019. Neonates with clinically diagnosed sepsis having blood culture positive were included in the study. Blood samples culture and antimicrobial susceptibility testing were performed with the standard microbiological method. Demographic, clinical information, and clinical outcomes were documented. The incidence of culture-positive sepsis was 10.3% (183/1773) of neonatal admissions. Poor feeding 85(46%) and fever 68(37%) were the common clinical features at presentation. The incidence of early-onset sepsis and late-onset sepsis were found to be 116 (63%) and 67(37%) respectively. Staphylococcus aureus was the common pathogen in both early-onset 61(49%) and late-onset 34(41%) sepsis. The incidence of multidrug-resistant cases was 41% (75/183) with 20% (15/75) extensively drug-resistant gram-negative bacilli, 36% (20/75) multidrug-resistant gram-negative bacilli, and 44% (33/75) Methicillin-resistant Staphylococcus aureus cases. In-hospital mortality rate was 12 (7%) with a higher frequency in multidrug-resistant sepsis 92% (11/12) than non- multidrug-resistant 8% (1/12). The median hospital days were longer in multidrug-resistant cases than non- multidrug-resistant [11(9-13) verses 3(2-5)]. The incidence of multidrug-resistant pathogens causing neonatal sepsis is high at our hospital and are associated with more in-hospital mortality and longer hospital stay. Implementation of effective preventive strategies to combat the emergence of antimicrobial resistance is immediately needed.

Identification of potential biogenic chalcones against antibiotic resistant efflux pump (AcrB) via computational study

The cases of bacterial multidrug resistance are increasing every year and becoming a serious concern for human health. Multidrug efflux pumps are key players in the formation of antibiotic resistance, which transfer out a broad spectrum of drugs from the cell and convey resistance to the host. Efflux pumps have significantly reduced the efficacy of the previously available antibiotic armory, thereby increasing the frequency of therapeutic failures. In gram-negative bacteria, the AcrAB-TolC efflux pump is the principal transporter of the substrate and plays a major role in the formation of antibiotic resistance. In the current work, advanced computer-aided drug discovery approaches were utilized to find hit molecules from the library of biogenic chalcones against the bacterial AcrB efflux pump. The results of the performed computational studies via molecular docking, drug-likeness prediction, pharmacokinetic profiling, pharmacophore mapping, density functional theory, and molecular dynamics simulation study provided ZINC000004695648, ZINC000014762506, ZINC000014762510, ZINC000095099506, and ZINC000085510993 as stable hit molecules against the AcrB efflux pumps. Identified hits could successfully act against AcrB efflux pumps after optimization as lead molecules. Communicated by Ramaswamy H. Sarma

High Frequency of Antiviral Resistance Mutations in HBV Genotypes A2 and H: Multidrug Resistance Strains in Mexico.

Lamivudine (3TC), telbivudine (LdT), entecavir (ETV), adefovir (ADF), and tenofovir (TFV) are drugs used to treat hepatitis B virus (HBV) infection, but specific mutations allow some viruses to become resistant to antiviral drugs or to acquire immune escape capacities. These mutations have not been thoroughly investigated in Mexico. This study aimed to estimate the prevalence of HBV antiviral resistance and escape mutations. This cross-sectional study analyzed 158 samples. HBV DNA was extracted, amplified, and sequenced in serum samples using the spin column method, PCR assay, and Sanger's sequencing, respectively. HBV genotypes were determined, and HBV mutations were tested using the Geno2pheno tool. Overall, 68.4% (108/158) of HBV patients were infected with genotype H, followed by G (11.4%, 18/158), A2 (10.8%, 17/158), F1b (6.9.0%, 11/158), D (1.9%, 3/158), and E (0.6%, 1/158), and 5.1% (8/158) had evidence of recombination. The prevalence of resistance mutations was 8.2% (13/158) and the most common combined mutation was rt180M+rt204V. Notably, we found the combinations rt180M+rt204V+rt173L (n=2) and rt180M+rt204V+rt202G (n=1) that confer multidrug resistance to 3TC, LdT, and ETV. Resistance mutations were found in genotypes A2 (11.8%, 2/17), and H (10.2%, 11/108), and escape mutations were detected in HBV genotypes A2 (11.8%, 2/17), H (10.2%, 11/108), F1b (9.1%, 1/11) and G (5.6%, 1/18). The highest prevalence of antiviral resistance mutations or escape mutations was detected in HBV genotypes A2 and H. The earliest cases of HBV multidrug resistance were detected in Mexico.

Identification and Characterization of a Vancomycin Intermediate-Resistant Staphylococcus haemolyticus Isolated from Guangzhou, China.

Staphylococcus haemolyticus is an opportunistic pathogen that belongs to coagulase-negative Staphylococci (CoNS). Increasing infection and multi-drug resistance cases caused by this strain have been reported and thus it poses a great health threat. The third-generation sequencing technology was performed on a S. haemolyticus SH-1 isolated from a clinical sample to analyze the drug resistance genes, which included vancomycin resistance related genes. In addition, antimicrobial susceptibility tests, transmission electron microscopy and Triton X-100 stimulated autolysis were conducted to understand its biological characteristics. The study shows that this clinical isolate is a vancomycin intermediate-resistant strain. Genome comparison also revealed that WalK(N70K) and WalK(R280Q) mutations may contribute to the vancomycin resistant phenotype. Besides, S. haemolyticus SH-1 exhibit common features of thicker cell wall and decreased autolytic activity. S. haemolyticus SH-1 with WalKR mutations shows typical characteristics of vancomycin resistant strains. Combining the genome features and biological properties, our findings may provide important information for the understanding of the molecular mechanism of S. haemolyticus to vancomycin intermediate-resistance.

Clinical value of the MeltPro MTB assays in detection of drug-resistant tuberculosis in paraffin-embedded tissues

Objective: To evaluate the clinical value of the MeltPro MTB assays in the diagnosis of drug-resistant tuberculosis. Methods: A cross-sectional study design was used to retrospectively collect all 4 551 patients with confirmed tuberculosis between January 2018 and December 2019 at Beijing Chest Hospital, Capital Medical University. Phenotypic drug sensitivity test and GeneXpert MTB/RIF (hereafter referred to as "Xpert") assay were used as gold standards to analyze the accuracy of the probe melting curve method. The clinical value of this technique was also evaluated as a complementary method to conventional assays of drug resistance to increase the detective rate of drug-resistant tuberculosis. Results: By taking the phenotypic drug susceptibility test as the gold standard, the sensitivity of the MeltPro MTB assays to detect resistance to rifampicin, isoniazid, ethambutol and fluoroquinolone was 14/15, 95.7%(22/23), 2/4 and 8/9,respectively; and the specificity was 92.0%(115/125), 93.2%(109/117), 90.4%(123/136) and 93.9%(123/131),respectively; the overall concordance rate was 92.1%(95%CI:89.6%-94.1%),and the Kappa value of the consistency test was 0.63(95%CI:0.55-0.72).By taking the Xpert test results as the reference, the sensitivity of this technology to the detection of rifampicin resistance was 93.6%(44/47), the specificity was100%(310/310), the concordance rate was 99.2%(95%CI:97.6%-99.7%), and the Kappa value of the consistency test was 0.96(95%CI:0.93-0.99). The MeltPro MTB assays had been used in 4 551 confirmed patients; the proportion of patients who obtained effective drug resistance results increased from 83.3% to 87.8%(P<0.01); and detection rate of rifampicin, isoniazid, ethambutol, fluoroquinolone resistance, multidrug and pre-extensive drug resistance cases were increased by 3.2%, 14.7%, 22.2%, 13.7%, 11.2% and 12.5%, respectively. Conclusion: The MeltPro MTB assays show satisfactory accuracy in the diagnosis of drug-resistant tuberculosis. This molecular pathological test is an effective complementary method in improving test positivity of drug-resistant tuberculosis.

Comparison of active tuberculosis case finding strategies for immigrants in South Korea: Epidemiology and cost-effectiveness analysis.

Tuberculosis (TB) is one of the serious infectious diseases in South Korea, with 49 new cases per 100,000 people and 629 multi-drug resistant (MDR) cases reported in 2020. TB is increasing among immigrants in S. Korea, and various TB case finding strategies are being performed for screening. We compared active case finding (ACF) with passive case finding (semi-PCF) across epidemiological characteristics and investigated a cost-effective strategy for screening immigrants for TB. ACF driven by non-governmental organizations and semi-PCF as part of the government's visa renewal process using CXR with additional acid-fast bacilli (AFB) smear and cultures were performed. Epidemiological parameters were compared between the two TB screening projects, and costs were collected. Cost-effectiveness was evaluated using a decision analysis model from the health system perspective. The primary outcome was incremental cost-effectiveness ratio (ICER) per averted TB case. Additional probabilistic sensitivity analysis was conducted. ACF (2.02%) showed a higher TB prevalence rate than semi-PCF (0.67%) on CXR. For subjects older than 60 years, the suspected TB rate on CXR was significantly higher in ACF (36.6%) than in semi-PCF (12.2%) (P<0.01). TB incidence among the family visa type was significantly higher in ACF (1.96%) than in semi-PCF (0.88%) (P < 0.0012). Costs for ACF ($666.92) were $20.784 higher than for semi-PCF ($646.13), but TB progression decreased by 0.02, resulting in an ICER of $948.18 per averted TB case. In sensitivity analysis, the indirect costs of ACF and semi-PCF had the highest impact on ICER. ACF found more TB cases than semi-PCF through CXR screening, and suspect cases with old age and family visa type were more common in ACF than in semi-PCF. ACF is cost-effective as a TB screening strategy for immigrants.

Genomic and geospatial epidemiology of Mycobacterium tuberculosis in Oman: first national insight using whole genome sequencing

Mycobacterium tuberculosis (MTB) is a global public health issue. Although Oman reduced the burden of tuberculosis (TB) by 85% in under 25 years, the annual incidence rate remains stagnant. Whole genome sequencing (WGS) is used to investigate the transmission dynamics of MTB complex. This study aimed to resolve traditional genotype clusters and exploring the geospatial distribution to understand the epidemiology of TB in Oman. Confirmed cases with spoligotyping clusters were randomly selected. WGS of 70 isolates were selected for final analysis. Correlation of epidemiological and geospatial data was conducted. A total of 233 cases were registered in 2021; 169 had confirmed growth, with an incidence rate of 5.2/100,000 population for 2021. A total of 70 genomes were analyzed, and five large clusters and three medium clusters were identified. The lineages L1, L2, L3, and L4 and several sublineages belonging to the Indo-Oceanic family and East African Indian family were identified as predominant in Oman. There were no multidrug-resistant cases identified. There is a considerable genetic variation among the strains in Oman. This predominance could be linked with the high percentage of non-national population, which represents different countries and frequent traveling to high TB burden countries. WGS combined with geospatial investigations of MTB are required to better understand the disease transmission in Oman, which will support TB elimination efforts.

The Evolution and Transmission Dynamics of Multidrug-Resistant Tuberculosis in an Isolated High-Plateau Population of Tibet, China.

On the Tibetan Plateau, most tuberculosis is caused by indigenous Mycobacterium tuberculosis strains with a monophyletic structure and high-level drug resistance. This study investigated the emergence, evolution, and transmission dynamics of multidrug-resistant tuberculosis (MDR-TB) in Tibet. The whole-genome sequences of 576 clinical strains from Tibet were analyzed with the TB-profiler tool to identify drug-resistance mutations. The evolution of the drug resistance was then inferred based on maximum-likelihood phylogeny and dated trees that traced the serial acquisition of mutations conferring resistance to different drugs. Among the 576 clinical M. tuberculosis strains, 346 (60.1%) carried at least 1 resistance-conferring mutation and 231 (40.1%) were MDR-TB. Using a pairwise distance of 50 single nucleotide polymorphisms (SNPs), most strains (89.9%, 518/576) were phylogenetically separated into 50 long-term transmission clusters. Eleven large drug-resistant clusters contained 76.1% (176/231) of the local multidrug-resistant strains. A total of 85.2% of the isoniazid-resistant strains were highly transmitted with an average of 6.6 cases per cluster, of which most shared the mutation KatG Ser315Thr. A lower proportion (71.6%) of multidrug-resistant strains were transmitted, with an average cluster size of 2.9 cases. The isoniazid-resistant clusters appear to have undergone substantial bacterial population growth in the 1970s to 1990s and then subsequently accumulated multiple rifampicin-resistance mutations and caused the current local MDR-TB burden. These findings highlight the importance of detecting and curing isoniazid-resistant strains to prevent the emergence of endemic MDR-TB. IMPORTANCE Emerging isoniazid resistance in the 1970s allowed M. tuberculosis strains to spread and form into large multidrug-resistant tuberculosis clusters in the isolated plateau of Tibet, China. The epidemic was driven by the high risk of transmission as well as the potential of acquiring further drug resistance from isoniazid-resistant strains. Eleven large drug-resistant clusters consisted of the majority of local multidrug-resistant cases. Among the clusters, isoniazid resistance overwhelmingly evolved before all the other resistance types. A large bacterial population growth of isoniazid-resistant clusters occurred between 1970s and 1990s, which subsequently accumulated rifampicin-resistance-conferring mutations in parallel and accounted for the local multidrug-resistant tuberculosis burden. The results of our study indicate that it may be possible to restrict MDR-TB evolution and dissemination by prioritizing screening for isoniazid (INH)-resistant TB strains before they become MDR-TB and by adopting measures that can limit their transmission.

Clinical settings, management and factors affecting outcomes in multi and extensively-drug resistant Pseudomonas endophthalmitis.

To report the clinical settings, management and factors affecting outcomes in multi-drug resistant (MDR) and extensively-drug resistant (XDR) Pseudomonas endophthalmitis. Retrospective, consecutive, non-comparative interventional case series. Cases of MDR and XDR Pseudomonas endophthalmitis from January 2012 to December 2020 treated at our tertiary eye care center were included. Data collected included clinical data, anatomic and functional outcome, isolated micro-organisms, and culture sensitivity. This study included 29 eyes of 29 patients with MDR/XDR Pseudomonas endophthalmitis. Mean age at presentation was 60.27 ± 14.9 years (median 63). Commonest clinical setting was acute post-operative endophthalmitis in 27 cases (93.1%). Concurrent corneal infiltrate was present in 11 eyes (37.9%). Initial intervention in 19 eyes (65.5%) was vitrectomy, 2 of which underwent endoscopic pars plana vitrectomy. Mean number of interventions was 3.34 ± 1.44 (median 4). Mean follow up was 3.25 ± 3.07 months (median 2). Sensitivity to ceftazidime was 48.28%. All isolates were sensitive to colistin. Mean visual acuity at last follow up in logMAR was 2.64 ± 1.48 (median 3.5). Seven eyes (24.13%) were NPL (nil perception of light) at the last follow up. Two eyes (6.9%) underwent evisceration. Nine eyes (31.03%) had a favourable anatomic and functional outcome. Eyes without a corneal infiltrate at presentation were found to have a favourable anatomic and favourable functional outcome (OR 11.91, P < 0.04, CI 1.08 to 130.93). Corneal involvement at presentation is associated with poorer outcomes in MDR and XDR Pseudomonas endophthalmitis. There is a potential role of higher newer antibiotics especially colistin in the management of these cases.

An overview of current strategies and future prospects in drug repurposing in tuberculosis

A large number of the population faces mortality as an effect of tuberculosis (TB). The line of treatment in the management of TB faces a jolt with ever-increasing multi-drug resistance (DR) cases. Further, the drugs engaged in the treatment of TB are associated with different toxicities, such as renal and hepatic toxicity. Different combinations are sought for effective anti-tuberculosis (anti-TB) effects with a decrease in toxicity. In this regard, drug repurposing has been very promising in improving the efficacy of drugs by enhancement of bioavailability and widening the safety margin. The success in drug repurposing lies in specified binding and inhibition of a particular target in the drug molecule. Different drugs have been repurposed for various ailments like cancer, Alzheimer’s disease, acquired immunodeficiency syndrome (AIDS), hair loss, etc. Repurposing in anti-TB drugs holds great potential too. The use of whole-cell screening assays and the availability of large chemical compounds for testing against Mycobacterium tuberculosis poses a challenge in this development. The target-based discovery of sites has emerged in the form of phenotypic screening as ethionamide R (EthR) and malate synthase inhibitors are similar to pharmaceuticals. In this review, the authors have thoroughly described the drug repurposing techniques on the basis of pharmacogenomics and drug metabolism, pathogen-targeted therapy, host-directed therapy, and bioinformatics approaches for the identification of drugs. Further, the significance of repurposing of drugs elaborated on large databases has been revealed. The role of genomics and network-based methods in drug repurposing has been also discussed in this article.