Diagnostic Tool For Tuberculosis Research Articles

Contribution of TB-LAM Urine Test to Tuberculosis Diagnosis in HIV-infected Adult Patients in Burkina Faso

Aims: With the aim of improving the diagnosis of tuberculosis in patients living with HIV, we set out to evaluate the contribution of the TB-LAM urine test to the diagnosis of tuberculosis in PLWHA in Burkina Faso, in real life. Methodology: The study was conducted in the Infectious Diseases Departments of the Yalgado OUEDRAOGO (CHU-YO), Souro SANOU (CHU-SS) and Bogodogo (CHU-B) University Hospitals. These 03 centers are among the most important reference centers in Burkina Faso. This was a descriptive and analytical cross-sectional study with prospective data collection over 12 months, from January 1, 2023 to December 31, 2023, and involved PLWH inpatients and outpatients. Data were collected using the KoBoCollect v1.30.1 application and analyzed using STATA 16 software. Binary uni and multivariate logistic regression with a statistical significance level of 0.05 was used to identify associated factors. Results: A total of 113 patients were included in the study. The mean age was 43 ± 1.35 years, with a sex ratio of 0.85. The TB-LAM urine test was positive in 76 patients, for a positivity rate of 67.26%. The diagnosis of tuberculosis was confirmed by bacteriology in accordance with the national tuberculosis management protocol in 18/113 (15.93%) patients. Bacilloscopy and the Xpert test performed on all biological fluids, combined with the TB-LAM urine test, enabled the diagnosis of 81/113 (71.68%) patients in the study. Patients already on antiretroviral therapy (ORa=0.06 [0.06-0.73], p=0.02) were less likely to test positive for urinary TB-LAM, and those with unexplained asthenia (ORa=15.76 [1.19-20.66], p=0.03) were more likely to test positive. Conclusion: The TB-LAM urine test, combined with other tuberculosis diagnostic tools, increases the diagnostic yield of tuberculosis in PLWHA.

Xpert MTB/RIF assay in the diagnosis of pulmonary tuberculosis in children in tertiary care setting in South India.

Xpert MTB/RIF is recommended for the diagnosis of tuberculosis (TB) in children. We determined the performance of Xpert MTB/RIF in the diagnosis of pulmonary TB in children. The characteristics of children influencing Xpert MTB/RIF positivity were explored. Children aged <15 years with symptoms suggestive of pulmonary TB were prospectively enrolled from 2013 to 2019. Two sputum/early morning gastric aspirate specimens were collected for examination by smear (fluorescence microscopy), Xpert MTB/RIF, and culture [Mycobacteria growth indicator tube (MGIT)/Lowenstein-Jensen (LJ) medium]. Diagnostic performance of Xpert MTB/RIF was evaluated using LJ and or MGIT culture positivity as the reference standard. Sensitivity, specificity with 95% confidence interval (CI) were calculated. Stratified analysis was done; P < .05 was considered statistically significant. Of the total 1727 enrolled children, 1674 (97%) with complete results for at least one sputum/gastric aspirate sample were analyzed. The sensitivity of Xpert MTB/RIF was 68.5% in sputum and 53.6% in gastric aspirate while the specificity was 99% for both. The sensitivity compared to smear was 68.5% vs. 33.7% (P < .001) and 53.6% vs. 14.5%; (P < .001) in sputum and gastric aspirate, respectively. The sensitivity of Xpert MTB/RIF was 23.9% with decision to treat as reference standard. Xpert MTB/RIF positivity was significantly influenced by sex, age, nutritional status, chest X-ray abnormality, TB infection status, and symptoms suggestive of TB. Xpert MTB/RIF as an upfront test compared to smear improves diagnosis of pulmonary TB in children yet the sensitivity is suboptimal. Newer TB diagnostic tools with improved sensitivity is warranted in children.

Sex-specific blood-derived RNA biomarkers for childhood tuberculosis

Confirmatory diagnosis of childhood tuberculosis (TB) remains a challenge mainly due to its dependence on sputum samples and the paucibacillary nature of the disease. Thus, only ~ 30% of suspected cases in children are diagnosed and the need for minimally invasive, non-sputum-based biomarkers remains unmet. Understanding host molecular changes by measuring blood-based transcriptomic markers has shown promise as a diagnostic tool for TB. However, the implication of sex contributing to disease heterogeneity and therefore diagnosis remains to be understood. Using publicly available gene expression data (GSE39939, GSE39940; n = 370), we report a sex-specific RNA biomarker signature that could improve the diagnosis of TB disease in children. We found four gene biomarker signatures for male (SLAMF8, GBP2, WARS, and FCGR1C) and female pediatric patients (GBP6, CELSR3, ALDH1A1, and GBP4) from Kenya, South Africa, and Malawi. Both signatures achieved a sensitivity of 85% and a specificity of 70%, which approaches the WHO-recommended target product profile for a triage test. Our gene signatures outperform most other gene signatures reported previously for childhood TB diagnosis.

Rapid and Highly Sensitive Detection of Mycobacterium tuberculosis Utilizing the Recombinase Aided Amplification-Based CRISPR-Cas13a System.

Tuberculosis (TB), a disease caused by Mycobacterium tuberculosis (MTB) infection, remains a major threat to global public health. To facilitate early TB diagnosis, an IS6110 gene-based recombinase aided amplification (RAA) assay was coupled to a clustered, regularly interspaced short palindromic repeats (CRISPR)-Cas13a fluorescence assay to create a rapid MTB detection assay (named RAA-CRISPR-MTB). Its diagnostic efficacy was evaluated for sensitivity and specificity through sequential testing of recombinant plasmids, mycobacterium strains, and clinical specimens. RAA-CRISPR detected IS6110 genes at levels approaching 1 copy/μL with pUC57-6110 as the template and 10 copies/μL with H37Rv as the template. There was no observed cross detection of non-tuberculosis mycobacteria (NTM) with either template. Furthermore, RAA-CRISPR testing of 151 clinical specimens yielded a diagnostic specificity rate of 100% and a diagnostic sensitivity rate of 69% that exceeded the corresponding Xpert MTB/RIF assay rate (60%). In conclusion, we established a novel RAA-CRISPR assay that achieved highly sensitive and specific MTB detection for use as a clinical TB diagnostic tool in resource-poor settings.

A DNA Methylation Signature from Buccal Swabs to Identify Tuberculosis Infection.

Tuberculosis (TB) is amongst the largest infectious causes of death worldwide and there is a need for a time- and resource-effective diagnostic method. In this novel and exploratory study, we show the potential of using buccal swabs to collect human DNA and investigate the DNA methylation (DNAm) signatures as a diagnostic tool for TB. Buccal swabs were collected from pulmonary TB patients (n= 7), TB exposed (n= 7), and controls (n= 9) in Sweden. Using Illumina MethylationEPIC array the DNAm status was determined. We identified 5644 significant differentially methylated CpG sites between the patients and controls. Performing the analysis on a validation cohort of samples collected in Kenya and Peru (patients, n=26; exposed, n=9; control, n=10) confirmed the DNAm signature. We identified a TB consensus disease module, significantly enriched in TB-associated genes. Lastly, we used machine learning to identify a panel of seven CpG sites discriminative for TB and developed a TB classifier. In the validation cohort the classifier performed with an AUC of 0.94, sensitivity of 0.92, and specificity of 1. In summary, the result from this study shows clinical implications of using DNAm signatures from buccal swabs to explore new diagnostic strategies for TB.

Epidemiology and diagnostic tools practice of spinal tuberculosis at Kariadi Hospital Semarang, Indonesia

Introduction: Spinal tuberculosis (TB) is one of the most common manifestations of extrapulmonary TB. In contrast to pulmonary TB, currently, there is no consensus for standard management of spinal TB particularly in diagnosis. This study aims to provide an overview of spinal TB from an epidemiological profile and diagnostic practice perspective at Kariadi Hospital, Semarang. Method: We collect spinal TB patient data retrospectively through medical records in Kariadi Hospital Semarang between the 2019 to 2022 period. The data regarding all demographic profile, clinical, imaging, and laboratory data. Result: A total of 40 patients were included in this study, 23 patients were diagnosed through histopathology, 32 patients were diagnosed using gene expert examination, and 17 patients by Acid Fast Bacillus Test. The highest age distribution was 37.5% in the 21-30-year age group, with thoracic level involvement in 70% of cases. The average age was 35.10 ± 14.10 with the main complaint being weakness of the limbs followed by complaints of back pain. The MRI images found were paravertebral abscess (67.5%), followed by vertebral body compression (27.5%), vertebral body destruction (20%), wedging compression and disc narrowing (20%), and kyphosis (12, 5%). Conclusion: Establishing the diagnosis of spinal TB at Kariadi Hospital is very diverse, so management is needed regarding the selection of spinal TB diagnostic tools to provide appropriate management.

Health service providers’ perspective on barriers and strategies to tuberculosis treatment adherence in Obuasi Municipal and Obuasi East District in the Ashanti region, Ghana: a qualitative study

Abstract Background Despite the substantial improvement in treatment success rate over the past two decades in Ghana, some districts in the country still record high rates of tuberculosis (TB) deaths and lost-to-follow-up. The high incidence of these adverse outcomes suggests that the TB programme is not performing well in these districts which could be due to some barriers in the programme implementation. This paper explored the perspectives of healthcare providers on the barriers to TB treatment adherence and the potential strategies to address them in two high-burden districts in the Ashanti Region of Ghana. Methods This study utilised an explorative qualitative study design among 16 purposively selected healthcare providers directly involved in TB care in the Obuasi Municipal and Obuasi East District. Key informant interviews were used to collect data, which were audio-recorded and transcribed word-for-word. The transcriptions were then imported into Atlas.ti version 8.4 software for thematic content analysis. Results Findings from this study revealed significant socioeconomic and individual barriers to TB treatment adherence. Key socioeconomic barriers included income insecurity, transportation cost, food insecurity, lack of public education on TB and frequent shortage of TB diagnostic tools. The individual barriers identified were misconception of TB causation, perceived full recovery after intensive phase treatment, inadequate patient privacy and noncompliance to treatment transfer protocol. The strategies recommended by healthcare providers to address these barriers were home-based treatment, provision of food incentives, frequent education on TB, and usage of reminder systems for follow-ups. Conclusion The barriers to TB treatment adherence from the healthcare providers’ perspective were mainly socioeconomic and individual. The suggested strategies offer actionable steps towards addressing these barriers. Thus, it is recommended that the TB programme and other supporting stakeholders take into account these barriers and adopt these strategies when planning for TB control to enhance treatment adherence and improve patient health outcomes.

Machine Learning for Prediction of Tuberculosis Detection: Case Study of Trained African Giant Pouched Rats.

Technological advancement has led to the growth and rapid increase of tuberculosis (TB) medical data generated from different health care areas, including diagnosis. Prioritizing better adoption and acceptance of innovative diagnostic technology to reduce the spread of TB significantly benefits developing countries. Trained TB-detection rats are used in Tanzania and Ethiopia for operational research to complement other TB diagnostic tools. This technology has increased new TB case detection owing to its speed, cost-effectiveness, and sensitivity. During the TB detection process, rats produce vast amounts of data, providing an opportunity to identify interesting patterns that influence TB detection performance. This study aimed to develop models that predict if the rat will hit (indicate the presence of TB within) the sample or not using machine learning (ML) techniques. The goal was to improve the diagnostic accuracy and performance of TB detection involving rats. APOPO (Anti-Persoonsmijnen Ontmijnende Product Ontwikkeling) Center in Morogoro provided data for this study from 2012 to 2019, and 366,441 observations were used to build predictive models using ML techniques, including decision tree, random forest, naïve Bayes, support vector machine, and k-nearest neighbor, by incorporating a variety of variables, such as the diagnostic results from partner health clinics using methods endorsed by the World Health Organization (WHO). The support vector machine technique yielded the highest accuracy of 83.39% for prediction compared to other ML techniques used. Furthermore, this study found that the inclusion of variables related to whether the sample contained TB or not increased the performance accuracy of the predictive model. The inclusion of variables related to the diagnostic results of TB samples may improve the detection performance of the trained rats. The study results may be of importance to TB-detection rat trainers and TB decision-makers as the results may prompt them to take action to maintain the usefulness of the technology and increase the TB detection performance of trained rats.

Rapid detection of Mycobacterium tuberculosis in sputum using CRISPR-Cas12b combined with cross-priming amplification in a single reaction.

In this study, we successfully established a new One-Pot method, named TB One-Pot, for detecting Mtb in sputum by combining CRISPR-cas12b-mediated trans-cleavage with cross-priming amplification (CPA). Our study evaluated the diagnostic performance of TB One-Pot in clinical sputum samples for tuberculosis. The findings provide evidence for the potential of TB One-Pot as a diagnostic tool for tuberculosis.

Qualitative Assessment of Challenges in Tuberculosis control in Public Health Facilities of Bule Hora Town, Ethiopia; Health Workers’ and Tuberculosis control Program Coordinators’ Perspectives, 2020

Introduction: The Millennium Development Goal target of halting and reversing TB incidence has been achieved on a global scale. Mortality from TB has fallen by 47% since 1990. Despite these gains, however, TB still remains a major worldwide health crisis. According to a recent WHO report, there where in 2018, an estimated 10 million people fell sick with tuberculosis (TB) worldwide. 5.7 million Men, 3.2 million women and 1.1 million children. Poor health systems pose many barriers to effective tuberculosis (TB) control. This study aimed at exploring health workers and TB control program coordinator’s perspectives on health systems challenges facing TB control in Bule Hora Town, Oromia Region, southern, Ethiopia. Objective: To identify challenges of health care workers and tuberculosis control program co-ordinator towards tuberculosis control at Public health facilities of Bule Hora town-2020. Method: Qualitative study was conducted at Public health facilities of Bule Hora town, Ethiopia. Four in-depth interviews with TB control program coordinators and two focus group discussions among 8 health workers were conducted. Purposive sampling was used to recruit study participants. Thematic analysis was used to identify and analyze main themes. Results: We identified that shortage of laboratory reagents, Shortages of TB diagnostic tools and intermittent supply of anti-TB drugs, shortage of trained and motivated health care staffs, Poor community TB mobilization and collaboration between health institutions were challenges facing the TB control program performance in the study area. Conclusion: Ensuring no proper supply of anti-TB drugs, laboratory reagents and Shortages of TB diagnostic tools to all health facilities is essential. Continuous updated training of health workers on standard TB care and data handling, developing and implementing a sound retention strategy to attract and motivate health professionals to work in study areas are necessary interventions to improve the TB control program performance in the study area.

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.

Barriers to childhood tuberculosis case detection and management in Cambodia: the perspectives of healthcare providers and caregivers

BackgroundDiagnosis and treatment of tuberculosis (TB) in children remain challenging, particularly in resource-limited settings. Healthcare providers and caregivers are critical in improving childhood TB screening and treatment. This study aimed to determine the barriers to childhood TB detection and management from the perspectives of healthcare providers and caregivers in Cambodia.MethodWe conducted this qualitative study between November and December 2020. Data collection included in-depth interviews with 16 healthcare providers purposively selected from four operational districts and 28 caregivers of children with TB and children in close contact with bacteriologically confirmed pulmonary TB residing in the catchment areas of the selected health centers. Data were analyzed using thematic analyses.ResultsMean ages of healthcare providers and caregivers were 40.2 years (standard deviation [SD] 11.9) and 47.9 years (SD 14.6), respectively. Male was predominant among healthcare providers (93.8%). Three-fourths of caregivers were female, and 28.6% were grandparents. Inadequate TB staff, limited knowledge on childhood TB, poor collaboration among healthcare providers in different units on TB screening and management, limited quality of TB diagnostic tools, and interruption of supplies of childhood TB medicines due to maldistribution from higher levels to health facilities were the key barriers to childhood TB case detection and management. Caregivers reported transportation costs to and from health facilities, out-of-pocket expenditure, time-consuming, and no clear explanation from healthcare providers as barriers to childhood TB care-seeking. Aging caregivers with poor physical conditions, lack of collaboration from caregivers, ignorance of healthcare provider's advice, and parent movement were also identified as barriers to childhood TB case detection and management.ConclusionsThe national TB program should further invest in staff development for TB, scale-up appropriate TB diagnostic tools and ensure its functionalities, such as rapid molecular diagnostic systems and X-ray machines, and strengthen childhood TB drug management at all levels. These may include drug forecasting, precise drug distribution and monitoring mechanism, and increasing community awareness about TB to increase community engagement.

A Far-Red Molecular Rotor Fluorogenic Trehalose Probe for Live Mycobacteria Detection and Drug-Susceptibility Testing.

Increasing the speed, specificity, sensitivity, and accessibility of mycobacteria detection tools are important challenges for tuberculosis (TB) research and diagnosis. In this regard, previously reported fluorogenic trehalose analogues have shown potential, but their green-emitting dyes may limit sensitivity and applications in complex settings. Here, we describe a trehalose-based fluorogenic probe featuring a molecular rotor turn-on fluorophore with bright far-red emission (RMR-Tre). RMR-Tre, which exploits the unique biosynthetic enzymes and environment of the mycobacterial outer membrane to achieve fluorescence activation, enables fast, no-wash, low-background fluorescence detection of live mycobacteria. Aided by the red-shifted molecular rotor fluorophore, RMR-Tre exhibited up to a 100-fold enhancement in M. tuberculosis labeling compared to existing fluorogenic trehalose probes. We show that RMR-Tre reports on M. tuberculosis drug resistance in a facile assay, demonstrating its potential as a TB diagnostic tool.

A Far‐Red Molecular Rotor Fluorogenic Trehalose Probe for Live Mycobacteria Detection and Drug‐Susceptibility Testing

AbstractIncreasing the speed, specificity, sensitivity, and accessibility of mycobacteria detection tools are important challenges for tuberculosis (TB) research and diagnosis. In this regard, previously reported fluorogenic trehalose analogues have shown potential, but their green‐emitting dyes may limit sensitivity and applications in complex settings. Here, we describe a trehalose‐based fluorogenic probe featuring a molecular rotor turn‐on fluorophore with bright far‐red emission (RMR‐Tre). RMR‐Tre, which exploits the unique biosynthetic enzymes and environment of the mycobacterial outer membrane to achieve fluorescence activation, enables fast, no‐wash, low‐background fluorescence detection of live mycobacteria. Aided by the red‐shifted molecular rotor fluorophore, RMR‐Tre exhibited up to a 100‐fold enhancement in M. tuberculosis labeling compared to existing fluorogenic trehalose probes. We show that RMR‐Tre reports on M. tuberculosis drug resistance in a facile assay, demonstrating its potential as a TB diagnostic tool.

IFNG and IFNGR1 polymorphisms are associated with tuberculosis: a case-control study.

Previous studies suggested that single-nucleotide polymorphisms (SNPs) of interferon gamma (IFNG) and its receptor IFNGR1 may be involved in the pathogenesis of tuberculosis (TB). We aimed to examine the association of IFNG gene polymorphisms with TB in the Tibetan population and use the machine learning method to establish a clinical prediction model of TB. A total of 613 TB patients and 603 healthy controls were selected for the study. Associations between SNPs and TB were analyzed using logistic regression, adjusted for sex and age. Clinical data and SNPs were integrated to construct a TB prediction model using random forest (RF) machine learning. For IFNG, rs1861494 CT was a protective factor against TB compared with TT genotype (p = 0.010). The rs1861494 C allele was a protective factor for TB (p = 0.010). For IFNGR1, the rs3799488 C allele reduced the risk of TB by 30% (p < 0.001). rs9376267 CT (p = 0.005) and TT (p = 0.001) genotypes were protective factors for TB. Compared with the rs1327475 GG genotype, the frequency of the GA genotype in the case group significantly differed from the controls (p = 0.013). rs2234711 GA (p < 0.001), AA (p < 0.001) genotype and A (p < 0.001) alleles were also associated with TB. Finally, five markers are identified using the RF model. The area under the curve (AUC) reaches 0.6 in the training set and 0.59 in the test set. Our study found that IFNG and IFNGR1 gene polymorphisms were associated with TB in a Tibetan population. The results also demonstrate the potential of clinical-SNPs as diagnostic tools for TB.

Understanding Mycobacterium tuberculosis complex in elephants through a One Health approach: a systematic review

BackgroundMycobacterium tuberculosis complex (MTC) that causes the chronic infectious disease- tuberculosis (TB), often presents with a complicated epidemiological pattern where the transmission chain may include humans, domestic animals and wildlife, including elephants. TB has been reported globally in both captive and wild elephants. The One Health approach might be the most effective way of understanding the shared MTC infection dynamics in captive and wild animals like Asian elephants. This systematic review accumulates evidence on occurrence, transmission pathways, and preventive measures of TB in elephants from a One Health perspective.ResultsThe prevalence of TB reported in elephant populations ranges from 0 to 23.33% and high prevalence’s are reported for elephants that are in close proximity to infected humans. The risk of elephant to human infection transmission increased significantly with exposure duration and contact with infected elephants. Some studies described the plausible TB transmission to captive elephants from other animals (wild and domestic), suggesting inter- and intra-species transmission. The results of this systematic review based on 27 relevant published works, suggest three overarching interrelated transmission pathways for M. tuberculosis infections in Asian elephants- i) humans and elephants, ii) other animals (wild or domestic) and elephants and iii) unclear sources of infection.ConclusionsThe progress made with new TB diagnostic tools provides multiple methods to choose from. However, lack of harmonization of TB testing in elephants and their human contacts remains a challenge to prevent TB in those animals. Routine TB screening among elephants and caretakers by setting up an occupational health program for early diagnosis of infection through combined efforts of public health, veterinary medicine, and occupational health experts is suggested. This implies the need for a One Health approach to elephant TB control. This review reveals the need for more research on Mycobacterium tuberculosis complex transmission pathways at the human-animal interface.

Diagnostic toolkit for tuberculosis: should we include urine lipoarabinomannan (LAM) detection in the WHO European Region?

Diagnostic toolkit for tuberculosis: should we include urine lipoarabinomannan (LAM) detection in the WHO European Region?

Tuberculosis Diagnosis in the Era of SARS-CoV-2

Tuberculosis (TB) continues to be a global public health emergency responsible for approximately 1.3 million deaths annually. Enduring the existing TB challenges, the emergence of “severe acute respiratory syndrome coronavirus 2” (SARS-CoV-2), a similar respiratory infection threatened the success of TB control over the past few years. Contemplating the irreversible damage of the human immunodeficiency virus (HIV), one of the leading immune-suppressive conditions, a similar or worst expected with this synergism: TB-HIV-SARS-CoV-2. Therefore, an integrated approach is much demanded before the impending revolution, "Next Global Pandemic". The advancement of molecular diagnostic techniques, blood transcriptomics uncovered the importance of studying the cross-talk between host and pathogens. RNA-sequencing is a high-throughput sequencing technique allowing detailed characterization of gene expression profiles. With the impact of SARS-CoV-2 on host immunity, pathogen-derived biomarker identification is more disease-specific and constrains individual variations faced during host biomarker identification. However, several technical hurdles are encountered during the study of intracellular pathogens like Mycobacterium tuberculosis. The development of advanced RNA-sequencing techniques to tackle the issues targeting the host and pathogen interactions is in their infancy and restricted to in-vitro studies. Few studies on serum exosomal RNA-sequencing of active and latent TB patients enlightened the path of TB biomarker discovery urging the necessity of more studies. Thus, this review will explicitly discuss the existing TB diagnostic tools to understand where we stand in TB diagnosis and the recent advancements in blood transcriptomics emphasizing the importance of targeting the pathogen-derived biomarkers as a potential source for future diagnostics.

Knowledge, Attitude, and Practices Regarding Childhood Tuberculosis Screening and Management among Healthcare Providers in Al-Medinah Al-Munawara, Saudi Arabia

Among infectious illnesses, tuberculosis (TB) has the highest fatality rate. The likelihood of infection spreading from patients to healthcare providers is two times higher than in the general population. The primary risk factor for contracting TB is close proximity to an infected individual prior to diagnosis. The research assessed health workers’ knowledge, attitude, and practice of TB infection control in Saudi Arabia. The study used a cross-sectional approach based on an online survey. The research was carried out throughout the months of May to July 2022 in Al Madinah Al-Munawara. The questionnaire included demographic’s and work variables, knowledge regarding pediatric TB etiology, transmission, symptoms, lymph node characteristics, and diagnostic criteria. Also, the participants’ perspectives and practice patterns on contact investigation, TB diagnostic tools, laboratory services, presumptive TB diagnostic referrals, and TB treatment were evaluated. The study included 558 healthcare workers (HCWs) from various specialists. HCWs exhibited sound knowledge, and positive attitudes in comparison to those with minimal experience in diagnosing TB in children TB as 31.2% had excellent knowledge, 22.6% good, 18.5% mediocre, and 27.8% bad. The majority of responders (49.1%) had a positive attitude, while only 33.2% had correct and full practice routines. Immediate action is required to increase HCW awareness, capacity, and skills to ensure an accurate diagnosis. In the identification of TB cases, certain crucial information gaps were discovered. This study also highlights the significance of clinical experience and frequent interactions with tuberculosis patients in clinical practice as a pathway to competency-based learning by practice. Keywords: healthcare workers, knowledge, tuberculosis, diagnosis, practice, KSA.

An electrochemical aptasensor for Mycobacterium tuberculosis ESAT-6 antigen detection using bimetallic organic framework.

Alabel-free electrochemical aptasensor is reportedfor sensitive detection of the 6-kDa early secreted antigenic target (ESAT-6). For the first time, the bimetallic organic framework (b-MOF) of Zr-MOF-on-Ce-MOF was decorated with nitrogen-doped graphene (NG) and applied as the matrix for electroactive toluidine blue (Tb) to form the NG@Zr-MOF-on-Ce-MOF@Tb nanohybrid. Theprepared nanohybrid with excellent hydrophilicity, dispersibility, and large specific surface exhibited significant electrochemical response. Thisnanohybrid could be directly used for anchoring ESAT-6 binding aptamers (EBA) through the interaction between the 5'-phosphate group (PO43-) of EBA and Zr4+ of Zr-MOF. The signal response before and after incubating the ESAT-6 antigen has beenevaluated by cyclic voltammetry at a scan rate of 100mVs-1 from - 0.7 to 0.3V (vs. SCE). Under optimal conditions, the proposed aptasensor displayed a wide linear range from 100fgmL-1 to 10ngmL-1 with a limit of detection (LOD) of12fgmL-1. The developed method showed good reproducibility with a relative standard deviation (RSD) of 2.27%. Theaptasensor showed favorable results in the analysis of the real samples. With these merits, the aptasensor has exceptional potential as a diagnostic tool for tuberculosis in clinical practice.