Tuberculosis Laboratory Research Articles

<p>Geographic Accessibility, Readiness, and Barriers of Health Facilities to Offer Tuberculosis Services in East Gojjam Zone, Ethiopia: A Convergent Parallel Design</p>

BackgroundTuberculosis (TB) remains a foremost global public health threat. Active TB control needs geographically accessible health facilities that have quality diagnostics, equipment, supplies, medicines, and staff.ObjectiveThis study aimed at assessing the geographic distribution, physical accessibility, readiness and barriers of health facilities for TB services in East Gojjam zone, Ethiopia.MethodsA convergent parallel design was applied using health facility and geographic data. Data on facility attributes, service availability and readiness were collected by inteviewing TB officers, laboratory heads and onsite facility visits. Coordinates of health facilities and kebele centroids were collected by GPS. We used ArcGIS 10.6 to measure Euclidean distance from each kebele centroids to the nearest health facility. Descriptive statistics were computed by using SPSS version 25. Barriers to TB service readiness were explored by in-depth interviews. NVivo12 was used to thematically analyze the qualitative data.ResultsThe overall TB health service coverage (THSC) was 23% (ranging: 10–85%). The mean distance from the nearest health facility was 8km (ranging: 0.5–16km). About 132 (32%) kebeles had poor geographic accessibility to TB services (over 10km distance from the nearest health facility) and had poor facility readiness. Although 114 (95%) health facilities offered at least one TB service, 44 (38.6%) of them had no sputum smear microscopy. The overall TB readiness index was 63.5%: first-line anti-TB drugs (97%), diagnostics (63%), trained staffs, diagnostic and treatment guidelines (53%) and laboratory supplies (41%). Lack of health workers (laboratory personnel), inadequate budget, poor management practice and TB program support, inadequate TB commodity suppliers, and less accessible geographic locations of health facilities were identified as barriers to TB service readiness distribution.ConclusionConsiderable proportion of the population in the study area have poor access to quality TB diagnostic services due to low THSC and poor facility readiness. Barriers to TB service availability and readiness were health system related. Regular refresher training of health workers on TB, creating mechanisms to attract laboratory personnel to work in the study area and scaling up of sputum smear microscopy services, establishing an efficient mechanism for procurement, distribution, utilization and reporting of TB commodity supplies, and good management practices are crucial to enhance TB service readiness in the study zone.

Quality of Inpatient Tuberculosis Health Care in High-Burden Resource-Limited Settings: Protocol for a Comprehensive Mixed Methods Assessment Study

BackgroundThe quality of care for tuberculosis (TB) is deficient in high-burden countries and urgently needs improvement. However, comprehensively identifying the required improvements is challenging. Providing high-quality TB care is an important step toward improving patients’ quality of life and decreasing TB morbidity and mortality. Effective tools for assessing the quality of TB services using international standards and guidelines can identify existing gaps in services and inform improvements to ensure high-quality inpatient TB services.ObjectiveThis study aimed to develop evaluation instruments for defining the quality of provision of TB services.MethodsTo assess quality of services in the largest TB hospital in Armenia, we developed instruments based on the Joint Commission International Accreditation Standards for Hospitals, International Standards for TB Care, TB Laboratories Bio-Safety Standards, and the World Health Organization framework for conducting TB program reviews. A mixed methods approach was utilized, triangulating quantitative (checklists) and qualitative (in-depth interviews) results. A scoring system and strengths, weaknesses, opportunities, and treats analysis was applied to detail results for each of the 122 standards assessed. A scaling approach was used to present overall performances of inpatient services for eight patient-centered functions and five organization management functions.ResultsOverall, 40 in-depth interviews and 91 checklists (21 observations, 16 policy papers, 20 staff qualification documents, and 34 medical records) were developed, utilized, and analyzed to explore practices of health care professionals, assess inpatient treatment experience of patients and their family members, evaluate facility environmental conditions, and define the degree of compliance to standards.ConclusionsThe effective comprehensive evaluation instruments and methods developed in this study for quality of inpatient TB services support the implementation of similar effective assessments in other countries. It may also become a platform to develop similar approaches for assessing ambulatory TB services in resource-limited countries.International Registered Report Identifier (IRRID)DERR1-10.2196/13903

Tuberculosis drug resistance in Canada: 2018.

Drug-resistant tuberculosis (TB) is a public health issue of global importance that poses a threat to TB control efforts. Canada conducts nationwide surveillance to monitor emerging drug resistance trends and document progress towards reaching the goal of TB elimination. To describe TB drug resistance trends across Canada from 2008-2018, with a focus on 2018, by drug resistance, geographic and demographic patterns. TB drug resistance data are captured through two independent surveillance systems managed by the Public Health Agency of Canada: Canadian Tuberculosis Laboratory Surveillance System (CTBLSS) and the Canadian Tuberculosis Reporting System (CTBRS). Data from these systems were analyzed and descriptive statistics were reported by resistance profile, place of residence (province), age groups, sex and country of birth. In 2018, 1,459 TB isolates underwent drug susceptibility testing, a 4.3% decrease from 2017. Resistance to any first-line drug was reported in 148 isolates (10.1%), compared to 123 (8.1%) in 2017. Of these, 121 were monoresistant, five were polyresistant, 21 were multidrug-resistant tuberculosis (MDR-TB) and one was extensively drug-resistant TB (XDR-TB). Drug resistance was reported in all provinces and territories except Prince Edward Island, Northwest Territories and Yukon. Among individuals younger than 15 years, very little TB drug resistance was detected. Among individuals aged 15 years and older, the distribution of TB drug resistance varied with no discernable trends. The proportion of drug resistance was slightly higher in females than in males. By origin, 10.7% of foreign-born TB cases reported between 2006 and 2016 were drug-resistant. Among the Canadian-born non-Indigenous cases, 9.3% were drug resistant; among Canadian-born Indigenous, 2.4% were drug resistant. In 2018, the proportion of isolates with TB drug resistance in Canada remained low and below global averages, with stable drug resistance, both geographically and demographically.

Pattern of primary tuberculosis drug resistance and associated risk factors at Dubai health authority in Dubai.

The objective of this study is to determine the initial drug resistance pattern among new tuberculosis (TB) cases and assess the extent of association with human immunodeficiency virus (HIV) and diabetes mellitus (DM). This is a retrospective analysis of 1116 clinical isolates were collected from patients who were newly diagnosed with TB at TB Laboratory between January 2016 and November 2019 and used for determining drug-resistance profiles against five first-line and five second-line anti-TB drugs; and the results were assessed the association between TB risk factors and primary drug resistance TB. Of the 1116 newly diagnosed TB patients, 193 (17.3%) showed resistance to at least one or more of the first-line drugs by different patterns, 105 (9.4%) showed resistance to one drug, 38 (3.40%) showed polyresistance, 50 (4.5%) showed multidrug resistant (MDR), and one patient had extensively drug resistant. Mono-resistance to isoniazid (INH), STR, pyrazinamide, and rifampicin were seen in 40 (3.6%), 33 (2.95%), 29 (2.59%), and 3 (0.3%) of isolates, respectively. INH showed the highest percentage of resistance among the patients. Of 1116 newly diagnosed TB patients, 256 (22.9%) were TB-DM cases and 135 (12.9%) were TB-no DM cases. The rates of drug resistance-TB 46/1116 (4.12%), monoresistance 25 (2.24%), polyresistance 9 (0.8%), and MDR 12 (1.07%) among TB-DM group were higher than TB-no DM group. Our study confirms that resistance to INH was the most common phenomenon. We found that diabetes was identified as a risk factor of TB drug resistance. We did not find a significant association between HIV co-infection and TB drug-resistance.

Ongoing tuberculosis transmission among second-generation Ethiopian immigrants.

Despite considerable efforts to control tuberculosis (TB) among Ethiopian immigrants in Israel, an outbreak of TB among second-generation Ethiopian immigrants that involved native Israelis occurred between January 2011 and December 2019. The aim of this article is to report on this outbreak and discuss the patient and health system barriers that led to its propagation. Overall, 13 culture-positive TB patients were diagnosed in this outbreak. An additional 36 cases with identical mycobacterium tuberculosis genotypes were identified through cross-checking with the National TB Laboratory Registry. Among the 32 close contacts of the index case, 18 (56.3%) reported for screening and treatment of latent TB infection (LTBI) was recommended for 11 (61.1%) of them. However, none completed treatment and eight eventually developed TB. Of the 385 close contacts identified in this outbreak, 286 (74.3%) underwent contact investigation, 154 (53.8%) were recommended LTBI treatment, but only 26 (16.9%) completed the treatment. Routine contact investigation and treatment practice measures failed to contain the cascade of infection and disease, leading to the spread of the infecting strain of TB. This report highlights the challenges to identify the high-risk group and address barriers to care among such a vulnerable population.

Resazurin microtitre plate assay and Sensititre® MycoTB for detection of Mycobacterium tuberculosis resistance in a high tuberculosis resistance setting.

BackgroundRapid diagnosis of drug-resistant Mycobacterium tuberculosis is a challenge in low-income countries. Phenotypic drug susceptibility testing using Sensititre® MycoTB assay and the resazurin microtitre plate assay (REMA) are relatively new innovative methods to determine drug susceptibility.ObjectivesThis study aimed to determine the performance of the Sensititre and REMA for M. tuberculosis drug susceptibility testing in a high-volume tuberculosis reference laboratory.MethodsA laboratory-based study was performed at the Inkosi Albert Luthuli Central Hospital Tuberculosis Laboratory from January 2014 to June 2015. The Sensititre® MycoTB plate and REMA were compared to the gold standard agar proportion method (APM) using 134 stored isolates.ResultsAgreement between the Sensititre® MycoTB plate and APM was observed with 98% sensitivity, 82% specificity, 94% positive and 93% negative predictive values of the Sensititre® MycoTB assay for the detection of rifampicin resistance and 97%, 96%, 99% and 88% for isoniazid resistance. Good categorical agreement between the REMA and the APM was observed among isolates with 89% sensitivity, 68% specificity, 89% positive and 68% negative predictive value for the detection of rifampicin resistance and 95%, 96%, 99% and 81% for isoniazid resistance. Results for the second-line drugs showed elevated minimum inhibitory concentrations for multidrug-resistant and extensively drug-resistant tuberculosis isolates.ConclusionThe REMA and Sensititre® MycoTB plate are attractive alternatives to the gold standard APM for the phenotypic detection of M. tuberculosis drug resistance.

Bacillus Calmette–Guérin strains with defined resistance mutations: a new tool for tuberculosis laboratory quality control

ObjectiveLaboratory quality control (QC) is essential to assess the reliability of tuberculosis diagnostic testing. To provide safe QC reagents for the detection of drug-resistant Mycobacterium tuberculosis, we generated antibiotic-resistant mycobacterial strains of attenuated virulence (M. bovis bacillus Calmette–Guérin (BCG)). MethodsSeven mono-resistant BCG strains were developed by introducing resistance-conferring mutations into wild-type BCG strains. Mutations were confirmed by dideoxynucleotide sequencing. Phenotypic resistance was quantified by microbroth dilution to determine the MIC90. The capacity of two commercial tests (GeneXpert TB/RIF and Genotype MTBDRplus) to detect resistance-conferring mutations was evaluated independently. ResultsOur panel included BCG strains with mutations in rpoB (S450L, I491F), katG (deletion at AA428), gyrA (D94G), rpsL (K43R) and Rv0678c (S63R). These mutations translated respectively into phenotypic resistance to rifampin (MIC ≥8 mg/L), isoniazid (MIC ≥8 mg/L), moxifloxacin (MIC 4 mg/L) and streptomycin (MIC ≥8 mg/L); the Rv0678c mutant showed decreased susceptibility to both clofazimine (MIC 4 mg/L) and bedaqualine (MIC 1 mg/L). GeneXpert (Cepheid) and Genotype MTBDRplus (Hain Lifesciences) both called the rpoB S450L strain rifampin-resistant and the I491F mutant rifampin-susceptible, as expected based on single nucleotide polymorphism positions. Likewise, MTBDRplus called the novel katG deletion mutant isoniazid susceptible despite phenotypic resistance. ConclusionBCG strains engineered to be mono-resistant to anti-tuberculosis drugs can be used as safe QC reagents for tuberculosis diagnostics and drug susceptibility testing.

Development and application of affordable SNP typing approaches to genotype Mycobacterium tuberculosis complex strains in low and high burden countries

The Mycobacterium tuberculosis complex (MTBC) comprises the species that causes tuberculosis (TB) which affects 10 million people every year. A robust classification of species, lineages, and sub-lineages is important to explore associations with drug resistance, epidemiological patterns or clinical outcomes. We present a rapid and easy-to-follow methodology to classify clinical TB samples into the main MTBC clades. Approaches are based on the identification of lineage and sub-lineage diagnostic SNP using a real-time PCR high resolution melting assay and classic Sanger sequencing from low-concentrated, low quality DNA. Thus, suitable for implementation in middle and low-income countries. Once we validated our molecular procedures, we characterized a total of 491 biological samples from human and cattle hosts, representing countries with different TB burden. Overall, we managed to genotype ~95% of all samples despite coming from unpurified and low-concentrated DNA. Our approach also allowed us to detect zoonotic cases in eight human samples from Nigeria. To conclude, the molecular techniques we have developed, are accurate, discriminative and reproducible. Furthermore, it costs less than other classic typing methods, resulting in an affordable alternative method in TB laboratories.

Introduction of quality management in a National Reference Laboratory in Germany

BackgroundHigh quality diagnostic services are crucial for tuberculosis (TB) diagnosis, treatment and control. A strong laboratory quality management system (QMS) is critical to ensuring the quality of testing and results. Recent initiatives to improve TB laboratory quality have focused on low and middle-income countries, but similar issues also apply to high-income countries.Methods and findingsUsing a multipronged approach reviews of facilities, equipment, processes (purchasing, pre-analytic, analytic and post-analytic), staff, health and safety, documentation, information management and organization based on the ISO 15189 and the twelve quality system essentials were conducted between October 2015 and January 2016 at the National TB Reference Laboratory in Germany. Outcome assessment included proportion of smear positive slides, proportion of contaminated liquid cultures and DNA contamination rates before and after implementation of QMS. The odds ratio for these outcomes was calculated using a before/after comparison. Reviews highlighted deficiencies across all twelve quality system essentials and were addressed in order of priority and urgency. Actions aimed at improving analytical quality, health and safety and information management were prioritised for initial implementation in parallel with each other. The odds ratio for a sample to be tested as microscopically positive increased by 2.08 (95%CI 1.41–3.06) comparing the time before with the time after implementation of quality managed fluorescence microscopy. Liquid culture contamination rates decreased from 23.6- 7.6% in April-July 2016 to <10% in November 2017-March 2018. The proportion of negative controls showing evidence of DNA contamination decreased from 38.2% in 2013 to 8.1% in 2017, the corresponding odds ratio was 0.14 (95%CI 0.07–0.29).ConclusionThis study showed marked improvement on quality indicators after implementation of a QMS in a National TB Reference Laboratory. The challenges and lessons learned in this study are valuable not just for high-income settings, but are equally generalizable to other laboratories.

BILATERAL CAVITARY LUNG ADENOCARCINOMA IN A YOUNG LADY: A RARE CASE REPORT WITH REVIEW OF LITERATURE

A mucinous variant of bronchioloalveolar carcinoma is described as a rare variant of invasive adenocarcinoma and is characterized radiologically by consolidation, groundglass opacity, and nodules. Here, we report the case of a young lady with multiple cavitary disease presented with respiratory failure and mimicking as tuberculosis, pneumonia, vasculitis but initial laboratory investigations and bronchoscopy couldn’t clinch the diagnosis. Later on, CT-guided biopsy confirmed the diagnosis of mucinous bronchoalveolar carcinoma. Immunohistochemistry for TTF-1, EGFR, ALK, ROS mutation had been sent but the patient requested to discharge, have been put on EGFR inhibitor Gefitinib 250 mg once a day considering adenocarcinoma, female, Asian origin but couldn’t turn-up to review further details. The clinician should be aware of the cavitary presentation of adenocarcinoma of lung even in younger patients where the most common differential is tuberculosis.

Strengthening Quality of Tuberculosis Laboratories Toward Accreditation in Viet Nam.

Early diagnosis of tuberculosis (TB) and multidrug-resistant TB (MDR-TB) is a priority for Viet Nam's National TB Control Programme. In many laboratories, quality systems are weak; few have attained accreditation. We implemented a structured training and mentoring program for TB laboratories and measured impact on quality. Six TB culture laboratories implemented the Strengthening TB Laboratory Management Towards Accreditation (TB SLMTA) program, consisting of three training workshops and on-site mentoring between workshops to support improvement projects. Periodic audits, using standardized checklists, monitored laboratories' progress toward accreditation readiness. At baseline, all six laboratories achieved a zero-star level. At exit, five laboratories attained three stars and another one star. Overall checklist scores increased by 44.2% on average, from 29.8% to 74.0%; improvements occurred across all quality system essentials. The program led to improved quality systems. Sites should be monitored to ensure sustainability of improvements and country capacity expanded for national scaleup.

The National Tuberculosis Control Programme of Liberia Laboratory Programme Performance.

Background Tuberculosis (TB) is a major public health problem in Liberia. Little is known about the TB laboratory performance of Liberia and the challenges after the 14 years of civil war which ended in 2003. The purpose of the study was to evaluate the TB laboratory performance of Liberia. Methods A cross-sectional study was conducted from 2014 to 2015. The study was conducted using quantitative data of TB case findings, sputum microscopy proficiency testing, and on-site assessment of sputum microscopy laboratories in Liberia. 80 laboratories participated in the proficiency testing. Besides, four years' (2012–2015) TB case finding data obtained from the National Leprosy and Tuberculosis Control Programme (NLTCP) were used to complement the study. The data were analysed using descriptive statistics. Results From the 80 TB sputum microscopy testing laboratories participating in proficiency testing, only 20 (25%) scored acceptable performance. 46 (58%) TB microscopy laboratories reported quantification errors for the proficiency panel slide 6 which was 3+. The national TB smear-positive cases notified were 4342 in 2012 but decreased to 3820 and 2448 in 2013 and 2014, respectively. The TB smear case detection rate showed an increase from 68% in 2010 to 78% in 2011 and a decrease to 60%, 57%, and 42% in 2012, 2013, and 2014, respectively. Conclusion Between 2010 and 2013, the NLTCP succeeded in increasing the number of TB sputum microscopy laboratories. At most of the TB microscopy sites, the TB laboratory quality system was not implemented. The NLTCP of Liberia should develop strategies to overcome its challenges in TB laboratory testing.

Analisis Pelatihan, Motivasi, Ketersediaan Alat Dan Pengawasan Penggunaan APD Pada Laboran Tb Di Puskesmas Kota Pekanbaru

Abstract&#x0D; &#x0D; Tuberculosis (TB) is a zoonotic disease, this disease can be transmitted from animals to humans. The cause of tuberculosis is Mycobacterium tuberculosis (Soedarto, 2009). Based on the Health Profile in Pekanbaru from 2015-2017, in five public health centers were found with high tuberculosis cases, there are Sidomulyo 16% (77,58); Umban Sari 10% (46,63); RI Sidomulyo 6% (29,51); Rumbai Bukit 6% (28,99); and Lima Puluh 5% (24,64). From the highest TB ​​case data in the city of Pekanbaru, it can also have an impact on the public health centers Laboratory staff. According to Tuberculosis Laboratory Service Standards (2014) Occupational Safety in culture laboratories and sensitivity tests must be in accordance with the procedures of security in laboratory facilities with a high risk of transmission. The purpose of this study was to analyze training, motivation, equipment support, supervision in the transmission of tuberculosis in laboratory assistants in Public Health Center. This type of research is descriptive qualitative, conducted in five health centers in Pekanbaru City in January to February 2019. Informants are laboratory staff, Public Health Centre leaders and Head of Administration. The number of informants was 15 people. The instrument is in the form of an interview guide and observation sheet. The results of this study prove that laboratory staff have received TB training, laboratory motivation in using a fairly high protection tool, the availability of self-protection equipment in the laboratory and supervision carried out by the Puskesmas quality team on laboratory staff. From the results of this study, it can be recommended for Public Health Centre leaders to increase special training on protection in the laboratory.

Costs and operation management of community outreach program for tuberculosis in tribal populations in India.

To evaluate costs of an active case finding (ACF) program with tuberculosis (TB) treatment delivery and monitoring, which targeted a rural tribal population in India. A time and motion study was conducted to evaluate operations and workload. Costs from the program perspective were assessed using both the bottom-up and top-down costing methods, exclusive of routine TB care costs. The impact of ACF on routine TB laboratory workloads was measured based on the changes in available staff time per smear at nine designated microscopy centers before and after program implementation. A majority (53.2%) of the community health-care worker's time was spent in traveling to communities, with an average of 22 TB patients (95% CI 19.14-24.94) seen per day per person. Costs (at 2015 $US rates) were US$1.85-US$2.42 per patient screened and submitting sputum, US$2.51-US$4.74 per person diagnosed with TB, and US$22.52-US$34.13 per TB patient completing treatment. Total smear volumes increased significantly after the ACF program, with more than a 15% reduction in available staff time per sputum smear test in most laboratories. This low-cost, ACF program has the potential to be highly cost-effective in addressing gaps in TB care problems in rural India.

A case of Mycobacterium tuberculosis laboratory cross-contamination

SettingA laboratory cross-contamination event was suspected because Mycobacterium tuberculosis was unexpectedly detected at a high incidence in the cultures of several clinical specimens at the National Hospital Organization, Tokyo National Hospital, Japan. ObjectiveTo describe a case of Mycobacterium tuberculosis laboratory cross-contamination. DesignWe reviewed the medical records of 20 patients whose clinical specimens were suspected to have been contaminated by Mycobacterium tuberculosis. Variable number of tandem repeat analysis with 15 loci, the Japan Anti-Tuberculosis Association-12, and three additional hyper-variable loci, was performed to identify the cross-contamination event. ResultsThe clinical, laboratory, and variable number of tandem repeat data revealed that the cross-contamination had possibly originated from one strongly positive specimen, resulting in false-positive results in 11 other specimens, including a case treated with anti-tuberculosis drugs. ConclusionClinical and laboratory data must be re-evaluated when cross-contamination is suspected and variable number of tandem repeat analysis should be used to confirm cross-contamination. Furthermore, original isolates should be stored appropriately, without sub-culturing and genotyping should be performed at the earliest possible for better utilization of variable number of tandem repeat for the identification of cross-contamination.

PO 8446 COLLABORATIVE TUBERCULOSIS RESEARCH AGENDA AT KEMRI CENTER FOR GLOBAL HEALTH RESEARCH, KISUMU, KENYA

BackgroundDeveloping countries grapple with inadequate funding amidst high burden of diseases. Africa is home to 9 of the 22 countries with a high tuberculosis (TB) burden and to 29 of the 41 countries with a high TB-HIV burden. Kenya is among the high TB and high TB-HIV burden countries. The Western Kenya region has the highest burden of TB and HIV. North-South partnerships are pivotal in ameliorating funding gaps in clinical research.MethodsWhile optimising existing infrastructure and organising programme support (i.e sensitisation and awareness creation, leading to study participant recruitment), from 2005 to date we conducted with multiple North-South collaborators capacity strengthening, TB prevalence survey, observational studies, operational research, and vaccine and drug trials.ResultsTB prevalence survey showed 600 cases per 100,000 population, TB epidemiological studies among adolescents and infants yielded 680 and 900/100,000 population respectively while 2 TB vaccine trials among infants and adults were conducted in Siaya. Three TB drug trials and a TB patient observational cum bio-bank study were concluded in Kisumu. KEMRI TB laboratory was upgraded from BSL2 to BSL3, was ISO-accredited in 2013, renewed in 2015 and 2017 and supports TB programme health facilities with retreatment specimens, supervision and mentorship. Over 25 operational TB studies grouped into community and case detection (increasing case detection), diagnostic and molecular (new diagnostic methods) and epidemiology studies (testing and monitoring cohorts for epidemiological questions) were implemented. Five PhDs, 9 Master’s, 2 Postgraduate Diplomas, 6 Bachelor’s degrees and 10 Diplomas have been supported. Siaya clinical research center was built while in Kisumu an adolescent clinic was constructed. This work involved 18 northern and 26 southern partners. Over 35 publications have been published out of these collaborations.ConclusionNorth-South collaborations provided funding, expertise and resources to harness research capacity of KEMRI; hence the need to foster a global networking culture.

OC 8718 WHAT DID WE LEARN FROM PANACEA 1 CLINICAL TRIALS?

BackgroundThe Pan African Consortium for the Evaluation of Anti-Tuberculosis Antibiotics (PanACEA) was designed to build clinical tuberculosis (TB) trial capacity whilst conducting clinical trials on novel and existing agents to shorten and simplify TB treatment. One of the objectives was to conduct, mentor and monitor observational and clinical studies at sites in 6 Sub-Saharan TB-endemic countries (Gabon, Kenya, South Africa, Tanzania, Uganda and Zambia)MethodsLearning through experience. All centres in the 6 countries self-assessed their requirements for capacity development in the following fields: a) clinical staff availability and experience; b) TB laboratory infrastructure and staff; c) safety laboratory infrastructure and staff; d) clinical site facilities and equipment; e) pharmacy facilities and staff; IT facilities; and f) overall training needs of site personnel.ResultsFrom March 2011 – June 2014, we conducted four epidemiological studies (characterising TB patient populations in preparation for future studies) and five phase II studies (GCP standard intervention trials).By working together in epidemiological and clinical trials, the sites identified their needs for resources and training as well as developing capabilities to perform independent large-scale TB clinical trials beyond PanACEA-initiated trials. Through the ReMoxTB study, for example, laboratories were brought to an international standard for safety and mycobacterial expertise. Furthermore, through developing skill-sets related to EBA studies, sites have since then attracted other sponsors for further studies.Sites could be mentored to perform GCP-compliant clinical TB trials that is built on sound physical infrastructure, training and strong on-site leadership.ConclusionThe learning-by-doing approach meant that staff could be trained whilst acquiring new core competencies and revealing operational gaps. Our experience of conducting TB trials within an environment of mentoring, networking and training has provided a platform for establishing future sustainable research centres that has capacities to conduct highly regulated studies.

Detection of tuberculosis laboratory cross-contamination using whole-genome sequencing

ObjectiveDetection of tuberculosis laboratory cross-contamination using whole-genome sequencing. MethodsA total of 22 M. tuberculosis strains with high genotypic homology from one hospital were collected during the drug resistance surveillance. Genome sequencing and epidemiological investigation were conducted to determine the occurrence of cross-contamination. ResultsThe pair wise comparison between the genomes in each cluster indicated that 15 (71.4%) of 21 strains with available genomic data had no SNP differences with at least one other strain within the same cluster. The analysis of the specimen collection time found that, among the 16 strains collected on the same day, 14 (87.5%) of them had no SNP differences with one another strain; meanwhile, among the strains within the same cluster whose SNP distance was 0, 93.3% (14/15) of them had the same collection time, suggesting that these findings were most likely caused by cross contamination. ConclusionA high proportion of M. tuberculosis strains with genotypic homology from the single institute that shared the same process time period was most likely caused by the cross contamination. Whole genome sequencing analysis can help to determine the occurrence of cross contamination.

Competitive fitness of Mycobacterium tuberculosis in vitro.

Background:While, bacteria resistance mutations can affect competitive fitness, given our multidrug-resistant (MDR) prevalence, we conducted this study to determine the impact of MDR on the competitive fitness of Mycobacterium tuberculosis (MTB) complex MDR strains. We conducted a cross-sectional study at the University Clinical Research Center (UCRC) from January to December 2017. New TB patients over aged of 18 were recruited at University teaching hospital and health reference centers of Bamako in USTTB Ethical committee approved protocols.Methods:MDR and drug-susceptible (wild-type [WT]) MTB strains (T1 and Beijing) and MTB H37Rv were competed on solid media in UCRC’s Tuberculosis Laboratory. Competitive and individual cultures were incubated for 14 days at 37°C with 7% CO2. Number of generation, generation time, and relative competitive fitness (W) of the strains were calculated. Data were analyzed with Epi-Info 7.1.5.2 software (CDC). P value was considered significant when it was <0.05. Scientific calculator (CS-82TL) was used for competitive fitness parameters calculations.Results:We performed 24 competitive cultures and 10 individual cultures. In individual cultures, strains’ generation number was for Beijing (WT: 4.60 and mutant MR: 4.40), T1 (WT: 2.69 and MR: 2.37), and H37Rv: 2.91. Generation number of WT strains was less than those of MDR strains in both individual and competitive culture. Relative competitive fitness was below 1 (W˂1) in 83.3%.Conclusion:MDR strains were less competitive than WT strains in 83.3% of cases. Resistant mutation impacts bacteria fitness.

Impact of Refresher Training on Knowledge Regarding Tuberculosis among Health Workers of West Bengal

Background: Tuberculosis(TB) is one of the leading causes of morbidity and mortality in India. Peripheral health workers in rural areas come in contact with TB suspects regularly during their home visits. The success of the Revised National Tuberculosis Control Programme (RNTCP) depends largely upon the involvement of these workers particularly Auxiliary Nurse Midwives (ANMs). Materials and Methods: All the ANMs and Health Assistants attached to a block primary health center (BPHC) of West Bengal were called for a refresher training on tuberculosis and RNTCP. Medical Officer – Tuberculosis Unit of the area impacted the training along with Senior Treatment Supervisor (STS) and Senior Tuberculosis Laboratory Supervisor (STLS) of the concerned Tuberculosis Unit (TU). The training was of one full day duration and conducted at the BPHC. The resource persons told them about the diagnosis and management of TB cases according to RNTCP guidelines. A pre-test questionnaire having 20 multiple choice and true/ false type questions were given to the participants. The same questionnaire was given to them after the training. The score of post-tests was compared with the pre-test questionnaire. Results: In total 34 health workers participated in the training. 44.1% of the participants were ANM and 38.2% were Health Assistants (Female). Rest were Health Assistants (Male). The mean (SD) duration of their service was 17.79 (1.8) years. 35.3% of them had a graduate degree. The mean (SD) post-test score was significantly better than the mean (SD) pre-test score, 16.53 (1.73) vs 9.76 (1.71). There was a significant improvement in score for 12 questions. Conclusion: With time the knowledge regarding TB and RNTCP is fading in health workers. Regular refresher training is required for the health workers to remain updated about tuberculosis.