Pediatric Tuberculosis Research Articles

Tuberculosis-targeted next-generation sequencing and machine learning: An ultrasensitive diagnostic strategy for paucibacillary pulmonary tuberculosis and tuberculous meningitis

BackgroundExisting diagnostic approaches for paucibacillary tuberculosis (TB) are limited by the low sensitivity of testing methods and difficulty in obtaining suitable samples. MethodsAn ultrasensitive TB diagnostic strategy was established, integrating efficient and specific TB targeted next-generation sequencing and machine learning models, and validated in clinical cohorts to test plasma cfDNA, cerebrospinal fluid (CSF) DNA collected from tuberculous meningitis (TBM) and pediatric pulmonary TB (PPTB) patients. ResultsIn the detection of 227 samples, application of the specific thresholds of CSF DNA (AUC = 0.974) and plasma cfDNA (AUC = 0.908) yielded sensitivity of 97.01 % and the specificity of 95.65 % in CSF samples and sensitivity of 82.61 % and specificity of 86.36 % in plasma samples, respectively. In the analysis of 44 paired samples from TBM patients, our strategy had a high concordance of 90.91 % (40/44) in plasma cfDNA and CSF DNA with both sensitivity of 95.45 % (42/44). In the PPTB patient, the sensitivity of the TB diagnostic strategy yielded higher sensitivity on plasma specimen than Xpert assay on gastric lavage (28.57 % VS. 15.38 %). ConclusionsOur TB diagnostic strategy provides greater detection sensitivity for paucibacillary TB, while plasma cfDNA as an easily collected specimen, could be an appropriate sample type for PTB and TBM diagnosis.

Shortened Treatment Regimens for Drug Sensitive TB.

Tuberculosis (TB) has remained a global health challenge despite the availability of effective anti-tubercular drugs and various treatment strategies. Apart from the complications related to TB disease per se, adverse effects of antitubercular therapy (ATT) also contribute to morbidity. In addition to the adverse effects, the long duration of the treatment regimen also reduces the patient's acceptability of ATT. The available "short-course treatment regimens" are still relatively long, thereby adversely affecting treatment compliance. There is a need for effective, safe, short and intensive regimens for TB which can reduce the treatment cost and adverse effects, thereby improving its acceptance. With the emergence of new evidence, the World Health Organization (WHO) has recently endorsed 4 mo short duration ATT regimen for non-severe, drug-sensitive cases of tuberculosis. Even in severe forms of disease like tubercular meningitis (TBM), trials are underway evaluating efficacy and safety of shorter regimens. Inclusion of fluroquinolones and rifapentine help shorten the regimens. These shortened regimens, however, need more close monitoring for adverse effects and may need to be converted to longer course if there is inadequate clinical response. Thus, shorter regimens for pediatric TB are likely to not only decrease the burden on patients and healthcare but also improve compliance and lower the side effects of the drugs due to prolonged exposure. This article reviews the current evidence and the guidelines pertaining to the shortened, intensive regimens for drug-sensitive tuberculosis.

Extrapulmonary and Drug-Resistant Childhood Tuberculosis: Unveiling the Disease to Adopt the Optimal Treatment Strategy.

Paediatric tuberculosis (TB) is a substantial threat among infectious diseases, particularly considering the high risk of extrapulmonary tuberculosis (EPTB), severe forms of the disease, and the spreading of drug-resistant strains. Describing the characteristics of children with EPTB and those with drug-resistant tuberculosis (DR-TB) and analysing the role of second-line drugs could facilitate the management of these cases. This retrospective study was conducted on 271 children diagnosed with active TB disease (44 EPTB cases, 9 DR-TB cases), originating from diverse geographic areas, who were referred to the infectious disease unit at Meyer Children's Hospital, Florence, Italy, from 2006 to 2022. In most patients, the management of therapies was complicated by the impossibility to obtain drug susceptibility testing (DST) results, which improved over the years: 17/154 (11.04%) children had DST results between 2006 and 2013, and 50/117 (42.73%, p < 0.001) between 2014 and 2022. Second-line drugs were not exclusively administered to DR-TB cases, but also to EPTB cases (20/44, 45.45%). Drugs were generally well tolerated; adverse events occurred in 13 children (13/271, 4.80%) and were generally mild and reversable. Therapies were successful in 267 children (98.52%) considered cured, while 4 (1.48%) presented sequelae. Both univariate and multivariate logistic regression analyses were conducted to investigate factors associated with EPTB, DR-TB, and second-line drugs administration. Originating from Asia emerged as a risk factor associated with both EPTB and DR-TB (p = 0.013 and p = 0.045, respectively). The introduction of GeneXpert tests has significantly improved TB diagnosis and the obtaining of DST results. The administration of second-line therapies should be limited primarily to DR-TB cases, but it is possible that these drugs may also be beneficial in selected EPTB cases.

Critical Review of Tuberculosis Diagnosis in Children from Papua New Guinea Presenting to Health Facilities in the Torres Strait Islands, Australia.

Paediatric tuberculosis can be challenging to diagnose, and various approaches are used in different settings. A retrospective review was conducted on Papua New Guinea (PNG) children with presumptive TB who presented for health care in the Torres Strait Islands, Australia, between 2016 and 2019. We compared diagnostic algorithms including the modified Keith Edwards TB Score, The Union Desk Guide, and the new World Health Organization (WHO) algorithm, with diagnostic practices used in the remote Torres Strait Islands. Of the 66 children with presumptive TB, 7 had bacteriologically confirmed TB. The majority (52%) were under 5 years (median age 61 months), and 45% were malnourished. There was moderate agreement across the diagnostic methods (K = 0.34; 95% CI 0.23-0.46), with the highest concordance observed between The Union Desk Guide and the WHO's algorithm (K = 0.61). Local TB physicians might have over-diagnosed presumed lymph node TB while under-diagnosing TB overall. Enhancing the precision and promptness of paediatric TB diagnosis using practical tools is pivotal to decrease TB-related child mortality, notably in isolated regions like the Torres Strait and the Western Province of PNG.

Sulam Project: Tuberculosis (TB) Health Education for Primary School Students as a community service collaboration between the Department of Biology FMIPA UNP and UPSI Malaysia

Tuberculosis (TB) is a global issue that is a priority in both Indonesia and Malaysia. Since being designated by WHO at 1993 as a global public health threat, Tuberculosis (TB) has become one of the top 10 leading causes of death worldwide. Recently, the number of Tuberculosis (TB) patients in Indonesia has increased, especially paediatric TB cases. In addition, Tuberculosis (TB) cases also increased in Malaysia. For the West Sumatra region, the highest number of TB patients is in Padang City. Since Tuberculosis (TB) is a disease that can affect anyone including children, the school environment as one of the places where children gather and interact has the opportunity to be a place for the spread of TB cases as well as having an important role in preventing TB transmission. However, in reality, the role of schools in early prevention efforts related to Tuberculosis (TB) disease in children, especially elementary school students, is still not optimal. In line with this, Tuberculosis (TB) disease is closely related to the human respiratory system which is one of the topics discussed in the human anatomy and physiology course in the MBKM activity between the Department of Biology FMIPA UNP and the Department of Biology UPSI. Therefore, it is necessary to make preventive efforts in the form of Tuberculosis prevention education. The results of community service are that several stages of community service activities have been carried out regarding Tuberculosis (TB) prevention education for elementary school students, namely the students involved are enthusiastic about the activities held, then pre-service biology teacher are also actively engaged in preparing and carrying out this activity and students' knowledge related to general knowledge, symptoms, their relationship with the respiratory system and prevention efforts are classified as good.

Assessment of geneXpert test for diagnosis of pediatric pulmonary tuberculosis

Introduction: Pediatric tuberculosis is an infectious disease and is a leading cause of death in Nepal. The incidence of pediatric TB is a key tool of ongoing transmission of tuberculosis within the community and vital indicator of disease control. Its diagnosis is very much challenging and delay in diagnosis leads to mortality.&#x0D; Objectives: This study was conducted to assess the utility of the gene Xpert test for diagnosis of pediatric pulmonary tuberculosis.&#x0D; Methodology: A hospital based prospective study done at Pediatric department of a teaching hospital, Nepalgunj, Banke conducted over a period of 1 year from 1 January 2021 to 31 December 2021 with presumptive diagnosis of pediatric pulmonary tuberculosis. Chi-square test was used to determine the association between categorical variables and a receiver operating curve (ROC) analysis was performed to compare the performance of gene Xpert test with other lab parameters like sputum/gastric lavage sample microscopy for acid fast bacilli, erythrocytic sedimentation rate, Tuberculin test (TT) and chest X-ray.&#x0D; &#x0D; Results: We enrolled a total of 294 patients in our study. Out of 294 samples 5.1% samples came out to be positive for gene Xpert for MTB. The sputum/gastric lavage samples for AFB showed both sensitivity and specificity of 100%. The tuberculin test showed a sensitivity of 80% and specificity of 37.3%. Chest X-ray showed a sensitivity of 66.7% and specificity of 43.7%. The area under the curve for acid fast bacilli in microscopy, tuberculin test and ESR was found to be 100% (95% CI: 100%-100%), 58.6% (95% CI: 44.9%-72.4%), and 52.6% (95% CI: 37%-68.3%) respectively. &#x0D; &#x0D; Conclusion: This study highlights the utility of gene Xpert as a good diagnostic tool for rapid diagnosis of pulmonary tuberculosis in pediatric population.

The socioeconomic burden of pediatric tuberculosis and role of child-sensitive social protection

BackgroundHouseholds of children with tuberculosis (TB) experience financial and social hardships, but TB-specific social protection initiatives primarily focus on adults.MethodsWe conducted a single-arm, pilot study of multi-component supportive benefits for children with pulmonary TB in Kampala, Uganda. At diagnosis, participants received in-kind coverage of direct medical costs, a cash transfer, and patient navigation. Caregivers were surveyed before diagnosis and 2 months into TB treatment on social and financial challenges related to their child’s illness, including estimated costs, loss of income and dissaving practices.ResultsWe included 368 children from 321 households. Pre-diagnosis, 80.1% of caregivers reported that their child’s illness negatively impacted household finances, 44.1% of caregivers missed work, and 24% engaged in dissaving practices. Catastrophic costs (> 20% annual income) were experienced by 18.4% (95% CI 13.7–24.0) of households. School disruption was common (25.6%), and 28% of caregivers were concerned their child was falling behind in development. Two months post-diagnosis, 12 households (4.8%) reported being negatively affected by their child’s TB disease (difference -75.2%, 95% CI -81.2 to -69.2, p < 0.001), with limited ongoing loss of income (1.6%) or dissavings practices (0.8%). Catastrophic costs occurred in one household (0.4%) at 2 months post-diagnosis.ConclusionsHouseholds face financial and social challenges prior to a child’s TB diagnosis, and child-sensitive social protection support may mitigate ongoing burden.

TB NOTIFICATION IN CHILDREN: THE EXPERIENCE OF TARGETED COMMUNITY DEMAND CREATION AMIDST INSECURITY IN IMO STATE, NIGERIA

This paper explores the challenges and successes of tuberculosis (TB) notification in children amidst a backdrop of insecurity in Imo State, Nigeria. Emphasizing targeted community demand creation strategies, the study investigates the intricate relationship between insecurity, community engagement, and pediatric TB detection. Drawing upon a comprehensive literature review, this research delves into the epidemiology, diagnosis, and treatment of TB in children, the ramifications of insecurity on healthcare access in Imo State, and effective community engagement strategies in healthcare initiatives. The literature findings underscore the substantial burden of TB among children and the distinct hurdles faced in environments characterized by insecurity. The prevailing insecurity in Imo State significantly disrupts healthcare services, leading to delays in TB diagnosis and treatment initiation. Nevertheless, the review identifies community engagement approaches as a promising avenue for bolstering TB notification rates in pediatric populations. These approaches empower local communities to actively participate in the identification and management of TB cases, ultimately contributing to the enhancement of pediatric TB notification. A conceptual framework, derived from amalgamated literature, elucidates the intricate interplay between insecurity, community engagement, and TB notification. This study highlights the pivotal role of community-driven interventions in mitigating the challenges posed by insecurity in Imo State, Nigeria, and presents actionable recommendations for the development of healthcare policies and programs tailored to the region. This research not only augments our comprehension of TB notification among children within insecure contexts but also provides valuable insights into the broader domain of community-based healthcare interventions. It underscores the exigency for customized strategies to address TB in pediatric populations in regions marked by insecurity, ultimately contributing to the amelioration of health outcomes for children in Imo State, Nigeria, and analogous settings&#x0D; Keywords: TB, Children, Community, Nigeria.

Screening for tuberculosis infection and effectiveness of preventive treatment among people with HIV in low-incidence settings.

To determine the yield of screening for latent tuberculosis infection (LTBI) among people with HIV (PWH) in low tuberculosis (TB) incidence countries (<10 TB cases per 100 000 persons). A systematic review and meta-analysis were performed to assess prevalence and predictive factors of LTBI, rate of TB progression, effect of TB preventive treatment (TPT), and numbers needed to screen (NNS). PubMed and Cochrane Library were searched for studies reporting primary data, excluding studies on active or paediatric TB. We extracted LTBI cases, odds ratios, and TB incidences; pooled estimates using a random-effects model; and used the Newcastle-Ottawa scale for bias. In 51 studies with 65 930 PWH, 12% [95% confidence interval (CI) 10-14] had a positive LTBI test, which was strongly associated with origin from a TB-endemic country [odds ratio (OR) 4.7] and exposure to TB (OR 2.9). Without TPT (10 629 PWH), TB incidence was 28/1000 person-years (PY; 95% CI 12-45) for LTBI-test positive versus 4/1000 PY (95% CI 0-7) for LTBI-test-negative individuals. Among 625 PWH (1644 PY) receiving TPT, 15 developed TB (6/1000 PY). An estimated 20 LTBI-positive individuals would need TPT to prevent one case of TB, and numbers NNS to detect LTBI or prevent active TB varied according to a-priori risk of LTBI. The relatively high prevalence of LTBI among PWH and the strong correlation with origin from a TB-endemic country support risk-stratified LTBI screening strategies for PWH in low-incidence countries and treating those who test positive.

Evaluating pediatric tuberculosis dosing guidelines: A model-based individual data pooled analysis.

The current World Health Organization (WHO) pediatric tuberculosis dosing guidelines lead to suboptimal drug exposures. Identifying factors altering the exposure of these drugs in children is essential for dose optimization. Pediatric pharmacokinetic studies are usually small, leading to high variability and uncertainty in pharmacokinetic results between studies. We pooled data from large pharmacokinetic studies to identify key covariates influencing drug exposure to optimize tuberculosis dosing in children. We used nonlinear mixed-effects modeling to characterize the pharmacokinetics of rifampicin, isoniazid, and pyrazinamide, and investigated the association of human immunodeficiency virus (HIV), antiretroviral therapy (ART), drug formulation, age, and body size with their pharmacokinetics. Data from 387 children from South Africa, Zambia, Malawi, and India were available for analysis; 47% were female and 39% living with HIV (95% on ART). Median (range) age was 2.2 (0.2 to 15.0) years and weight 10.9 (3.2 to 59.3) kg. Body size (allometry) was used to scale clearance and volume of distribution of all 3 drugs. Age affected the bioavailability of rifampicin and isoniazid; at birth, children had 48.9% (95% confidence interval (CI) [36.0%, 61.8%]; p < 0.001) and 64.5% (95% CI [52.1%, 78.9%]; p < 0.001) of adult rifampicin and isoniazid bioavailability, respectively, and reached full adult bioavailability after 2 years of age for both drugs. Age also affected the clearance of all drugs (maturation), children reached 50% adult drug clearing capacity at around 3 months after birth and neared full maturation around 3 years of age. While HIV per se did not affect the pharmacokinetics of first-line tuberculosis drugs, rifampicin clearance was 22% lower (95% CI [13%, 28%]; p < 0.001) and pyrazinamide clearance was 49% higher (95% CI [39%, 57%]; p < 0.001) in children on lopinavir/ritonavir; isoniazid bioavailability was reduced by 39% (95% CI [32%, 45%]; p < 0.001) when simultaneously coadministered with lopinavir/ritonavir and was 37% lower (95% CI [22%, 52%]; p < 0.001) in children on efavirenz. Simulations of 2010 WHO-recommended pediatric tuberculosis doses revealed that, compared to adult values, rifampicin exposures are lower in most children, except those younger than 3 months, who experience relatively higher exposure for all drugs, due to immature clearance. Increasing the rifampicin doses in children older than 3 months by 75 mg for children weighing <25 kg and 150 mg for children weighing >25 kg could improve rifampicin exposures. Our analysis was limited by the differences in availability of covariates among the pooled studies. Children older than 3 months have lower rifampicin exposures than adults and increasing their dose by 75 or 150 mg could improve therapy. Altered exposures in children with HIV is most likely caused by concomitant ART and not HIV per se. The importance of the drug-drug interactions with lopinavir/ritonavir and efavirenz should be evaluated further and considered in future dosing guidance. ClinicalTrials.gov registration numbers; NCT02348177, NCT01637558, ISRCTN63579542.

Pediatric Drug-Resistant Tuberculosis: The Current and Future Prospects for Management and Prevention.

In the continued battle against one of the oldest enemies known to mankind, Mycobacterium tuberculosis (MTB), the emergence of drug resistance to antituberculosis drugs among children poses multiple challenges for early detection and treatment. Molecular diagnostics and newer drugs like bedaquiline and delamanid have strengthened the armamentarium and helped design convenient, safe, and child-friendly therapeutic regimens against drug-resistant tuberculosis (TB). Preventive strategies like treatment of TB infection among children living in close contact with patients with drug-resistant TB and effective vaccines against TB are currently in the investigative stages of development and implementation. In addition to the implementation of recent novel diagnostics and treatment modalities, effective psychosocial and nutritional support, as well as dedicated monitoring for compliance and adverse effects, are crucial determinants for successful treatment outcomes in these children.

CLINICAL PROFILE AND OUTCOME OF PEDIATRIC TUBERCULOSIS IN A TERTIARY CARE SETTING IN CENTRAL INDIA

Objective: To study the clinical profile and outcome of tuberculosis (TB) in children under 14 years of age. Methods: It is a hospital-based cross-sectional study done in a tertiary referral center in Central India from July 2017 to June 2019. After ethical approval, 80 children (under 14 years of age) diagnosed with presumptive TB were enrolled. Those already on TB treatment or prophylaxis for more than 7 days were excluded from the study. Relevant patient details were noted on a predesigned pro forma. Routine first-line investigations for TB were done in all cases. Histopathological and specific radiology tests were done as per the site involved. Microbiological confirmation was done using microscopy, and molecular diagnosis was done by a cartridge-based nucleic acid amplification test (CBNAAT). Results: Overall mortality was 13.8%, and 82% of them were children below 5 years of age. As per WHO criteria, 52.5% and 20% of children below 5 years of age were severely and moderately undernourished, respectively. History of contact was present in 66%; BCG scar in 91.3%; and tuberculin sensitivity test positivity in 56% of cases. Fever, cough, and weight loss were the most common presenting complaints. The majority had extrapulmonary involvement (75%), with neurotuberculosis being the most common and with the highest mortality (70%). Microbiological confirmation was possible in only 8.8% of cases. Conclusion: It is still challenging to diagnose pediatric TB. Though newer diagnostics are now available, clinical suspicion is a valuable tool. The diagnosis of pediatric TB should thus be based on a combination of epidemiological variables, clinical suspicion, and supported by various laboratory investigations.

Household costs incurred when seeking and receiving paediatric tuberculosis services: a survey in Cameroon and Kenya

Background Elimination of catastrophic costs due to tuberculosis (TB) is one of the three targets of the World Health Organization (WHO) End TB Strategy. Limited data have yet been reported on the costs experienced by households of children receiving TB services. We quantified the economic impact on households with children seeking and receiving TB services during the Catalyzing Pediatric TB Innovations (CaP-TB) project in Cameroon and Kenya. Methods Within the INPUT stepped-wedge cluster-randomised study evaluating the effect of CaP-TB integration of TB services in paediatric entry points, we designed a cross-sectional facility-based survey with retrospective data collection using a standardised questionnaire adapted from the WHO Global taskforce on TB patient cost generic survey instrument. Caregivers of children receiving TB services (screening, diagnosis and treatment of drug-sensitive TB) during the CaP-TB project were interviewed between November 2020 and June 2021. Direct medical, direct non-medical, and indirect costs for TB services were analysed following WHO Global taskforce recommendations. We used the human capital and output-based approaches to estimating income loss. All costs are presented in 2021 US dollars. Results A total of 56 caregivers representing their households (Cameroon, 26, and Kenya, 30) were interviewed. The median household costs for TB services, estimated using the human capital approach, were $255 (IQR; $130-631) in Cameroon and $120 ($65-236) in Kenya. The main cost drivers across both countries were direct non-medical costs (transportation and food), 52%; and medical costs, 34%. Approximately 50% of households reported experiencing dissavings (taking a loan, or selling an asset) to deal with costs related to TB disease. Using a threshold of 20% of annual household income, 50% (95%CI; 37-63%) of households experienced catastrophic costs when using the human capital approach; (46% (95%CI; 29-65%) in Cameroon and 53% (95%CI; 36-70%) in Kenya). Estimated costs and incidence of catastrophic costs increased when using the output-based approach in a sensitivity analysis. Conclusions Accessing and receiving TB services for children results in high levels of cost to households, despite the provision of free TB services. Strategies to reduce costs for TB services for children need to address social protection measures or explore decentralisation. Registration: https://clinicaltrials.gov/ct2/show/NCT03862261.

Does multiple gastric aspirate collection increase sensitivity of M. tuberculosis detection in children with pulmonary tuberculosis?

This study aims to investigate the sensitivity of microscopy, culture and polymerase chain reaction on three gastric aspirates (GAs) in the microbiological confirmation of active pulmonary tuberculosis (TB) and to identify possible changes in sensitivity derived from the collection of a different number of aspirates. Children with clinical and radiological diagnoses of active pulmonary TB who underwent three GAs between March 2007 and June 2019 were retrospectively evaluated. Clinical, radiological, and microbiological data were collected. The sensitivity of microbiological tests on GAs was calculated. Moreover, differences in sensitivity according to age and radiological pattern were investigated. Overall, 156 children with active pulmonary TB were enrolled with a median age of 51.5 (IQR: 25.2–113.2) months. Microbiological investigations on the first GA showed a sensitivity of 34% (95%CI 26.7, 42), the cumulative sensitivity of first and second GAs was 40.4% (95%CI 32.7, 48.5) and of the three GAs was 47.4% (95%CI 39.8, 55.2). The collection of three GAs leads to an overall increase in sensitivity of the first GA by 13.4% (95%CI 2.8, 24.1%; p=0.014). Moreover, the increase in sensitivity was significantly higher in children ≤ 4 years of age and in those with uncomplicated TB (p=0.008).Conclusions: Performing a higher number of GAs increases the sensitivity of microbiological confirmation of active pulmonary TB, particularly in children ≤ 4 years and with an uncomplicated radiological pattern.What is known:• The diagnosis of paediatric tuberculosis is a challenge for paediatricians• Despite their low sensitivity gastric aspirates represent the standard sample for microbiological confirmation of active pulmonary tuberculosis in children• Most international guidelines recommend performing three sequential gastric aspirates on three consecutive daysWhat is new:• A significant increase in global sensitivity by 13.4% was found by the collection of three gastric aspirates compared to the first one• Performing a higher number of gastric aspirates increases the sensitivity of microbiological confirmation, particularly in children ≤ 4 years and with an uncomplicated radiological pattern

Tuberculosis in Spain: An opinion paper.

This document is the result of the deliberations of the Committee on Emerging Pathogens and COVID-19 of the Illustrious Official College of Physicians of Madrid (ICOMEM) regarding the current situation of tuberculosis, particularly in Spain. We have reviewed aspects such as the evolution of its incidence, the populations currently most exposed and the health care circuits for the care of these patients in Spain. We have also discussed latent tuberculosis, the reality of extrapulmonary disease in the XXI century and the means available in daily practice for the diagnosis of both latent and active forms. The contribution of molecular biology, which has changed the perspective of this disease, was another topic of discussion. The paper tries to put into perspective both the classical drugs and their resistance figures and the availability and indications of the new ones. In addition, the reality of direct observation in the administration of antituberculosis drugs has been discussed. All this revolution is making it possible to shorten the treatment time for tuberculosis, a subject that has also been reviewed. If everything is done well, the risk of relapse of tuberculosis is small but it exists. On the other hand, many special situations have been discussed in this paper, such as tuberculosis in pediatric age and tuberculosis as a cause for concern in surgery and intensive care. The status of the BCG vaccine and its present indications as well as the future of new vaccines to achieve the old dream of eradicating this disease have been discussed. Finally, the ethical and medicolegal implications of this disease are not a minor issue and our situation in this regard has been reviewed.

ВОЗРАСТНЫЕ ОСОБЕННОСТИ ФОРМИРОВАНИЯ ВНЕКЛЕТОЧНЫХ ЛОВУШЕК НЕЙТРОФИЛОВ У ЗДОРОВЫХ ЛИЦ И БОЛЬНЫХ ТУБЕРКУЛЁЗОМ

Abstracts. Introduction. Forming a neutrophil extracellular trap (NET) from DNA (NETosis) is considered as a potentially significant marker in the diagnosis of tuberculosis. However, there is practically a lack of data regarding the age-related differences in forming extracellular traps in research publications. Aim. To determine the age-related differences of NETosis in healthy individuals and in tuberculosis (TB) patients. Materials and Methods. The ability of neutrophils to form extracellular traps was investigated in 4 groups: 1) 4-7-year-old children diagnosed with the primary tuberculosis complex, who fell ill within Omsk region; 2) healthy and uninfected children at the same age; 3) adults with tuberculosis, and 4) adults without tuberculosis. For this purpose, isolated fractions of neutrophils from peripheral blood of the groups under research were stimulated with a mixture of bacteria (probiotic) for 30 minutes. In the hanging-drop preparation, we analyzed the proportion of neutrophil extracellular traps and cells in the state of various motility degrees. Results and Discussion. Age-related differences in the ability of neutrophils to form extracellular traps were characterized by a more pronounced response of neutrophils to the NETosis stimulant in children in the control group in comparison with neutrophils isolated from blood in the group of healthy adult volunteers: Neutrophils of children (Me=16.1; Q1=12.8; Q3=18.3) were statistically significantly more intense (p=0.0002) than those of adults (Me=8.6; Q1=7.1; Q3=10.5) and formed filamentous NETs. In the groups of pediatric and adult pulmonary TB patients, the response of neutrophils to the NETosis stimulant was diametrically opposite. Neutrophils of children with primary tuberculosis complex, as of those in the control group, predominantly formed filamentous NETs, but in greater amounts, while neutrophils of adults with infiltrative or focal form and with at most 2 segments involved mainly formed cloudy NETs, whereas filamentous NETs prevailed for the adults in the control group after stimulation. At the same time, proportion of cloudy NETs did not exceed 5% in healthy children and healthy adults. Comparing the findings of our study with literature data, we hypothesized that, most likely, there is a greater increase in PAD4 expression and more pronounced histone H3 citrullination in children’s neutrophils than in those of adults. Conclusions. Our findings indicate more pronounced ex vivo processes induced by NETosis stimulator in both groups of children (healthy individuals and TB patients) as compared to adults in the respective groups, which should be considered by the researchers interpreting the findings of the ability of neutrophils to form extracellular traps in tuberculosis.

Pediatric sellar-suprasellar tuberculosis: A case report and review of the literature

Background: Pediatric sellar-suprasellar tuberculosis is a rare form of tuberculosis that affects the pituitary gland and surrounding areas in the brain. It can be difficult to diagnose based on clinical and radiological signs alone, as they can be similar to other pituitary masses. A combination of biological, hormonal, and imaging examinations can aid in making an accurate diagnosis. It is important to consider tuberculosis in the differential diagnosis of sellar-suprasellar masses in the pediatric population, especially in areas with a high prevalence of tuberculosis. Case Description: A 17-year-old male with no history of illness showed up with a series of symptoms, including headaches and vision problems. A sellar-suprasellar lesion was seen on imaging, along with several minor lesions. The diagnosis of tuberculosis meningitis with cerebral and pituitary tuberculoma was made after cerebrospinal fluid analysis revealed the presence of tuberculosis. Treatment with anti-tuberculosis drugs led to clinical improvement and lesion resolution. Conclusion: Children’s sellar tuberculomas can be difficult to diagnose since they resemble other pituitary tumors. It is essential to take them into account in the differential diagnosis, especially in regions with a high incidence of tuberculosis. Long-term chemotherapy is the recommended course of treatment, and monthly follow-up visits are necessary to check hormone levels and evaluate whether a permanent hormone replacement is necessary.

More Than a Decade of GeneXpert®Mycobacterium tuberculosis/Rifampicin (Ultra) Testing in South Africa: Laboratory Insights from Twenty-Three Million Tests.

This study seeks to describe the rollout and current state of South Africa's GeneXpert molecular diagnostic program for tuberculosis (TB). Xpert MTB/RIF was introduced in 2011 with a subsequent expansion to include extra-pulmonary and paediatric testing, followed by Xpert MTB/RIF Ultra in 2017. Through a centralised laboratory information system and the use of a standardised platform for more than a decade, over 23 million tests were analysed, describing the numbers tested, Mycobacterium tuberculosis complex detection, rifampin resistance, and the unsuccessful test rates. The stratification by province, specimen type, age, and sex identified significant heterogeneity across the program and highlighted testing gaps for men, low detection yield for paediatric pulmonary TB, and the effects of inadequate specimen quality on the detection rate. The insights gained from these data can aid in the monitoring of interventions in support of the national TB program beyond laboratory operational aspects.

Diagnostic Utility of Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) on Induced Sputum Versus Gastric Aspirate Samples for the Diagnosis of Paediatric Pulmonary Tuberculosis.

Tuberculosis (TB) in children is neglected, mainly due to a lack of sensitive diagnostic tools. Paediatric TB is now a global priority. More paediatric TB cases are being recorded as a result of the introduction of Xpert® Mycobacterium tuberculosis (MTB)/rifampicin (RIF) (Cepheid Inc., Sunnyvale, USA). This study was undertaken to evaluate the performance of Xpert MTB/RIF in the diagnosis of pulmonary TB in children. We recruited 70 paediatric patients with probable pulmonary TB and their gastric aspirate (GA), and induced sputum (IS) samples were collected between January 2021 and June 2022 in Saifai, Etawah, Uttar Pradesh, at the Microbiology Department of the Uttar Pradesh University of Medical Sciences (U.P.U.M.S.). All samples were subjected to smear examination, Bacterial Activation ofContinuousTemperature andEnvironmentalControl -MycobacterialGrowthIndicatorTube (BACTEC-MGIT) culture, and Xpert MTB/RIF. The specimens included 70 GAs and 70 IS samples. The total number of specimens were 140 and we collected GA as well as IS from each of the patient enrolled in the study.When compared to microscopy, GeneXpert provides a quicker and earlier detection of paediatric TB. The sensitivity of the cartridge-based nucleic acid amplification test (CBNAAT) against mycobacterial growth indicator tube (MGIT) was 75.0% for GA samples and 63.64% for IS samples. Paediatric TB, owing to its paucibacillary nature and difficulty in the collection of samples, makes the diagnosis difficult by conventional methods. Our study shows that smear and culture yield in GA samples are superior to those of IS samples and the sensitivity of Xpert MTB/RIF assay is also significantly different in GA and IS samples, but a combination of GA and IS yielded the best results.

The diagnostic yield of nasopharyngeal aspirate for pediatric pulmonary tuberculosis: a systematic review and meta-analysis

BackgroundTuberculosis (TB) is a leading cause of death in children, but many cases are never diagnosed. Microbiological diagnosis of pulmonary TB is challenging in young children who cannot spontaneously expectorate sputum. Nasopharyngeal aspirates (NPA) may be more easily collected than gastric aspirate and induced sputum and can be obtained on demand, unlike stool. However, further information on its diagnostic yield is needed.MethodsWe systematically reviewed and meta-analyzed the diagnostic yield of one NPA for testing by either culture or nucleic acid amplification testing (NAAT) to detect Mycobacterium tuberculosis from children. We searched three bibliographic databases and two trial registers up to 24th November 2022. Studies that reported the proportion of children diagnosed by NPA compared to a microbiological reference standard (MRS) were eligible. Culture and/or WHO-endorsed NAAT on at least one respiratory specimen served as the MRS. We also estimated the incremental yield of two NPA samples compared to one and summarized operational aspects of NPA collection and processing. Univariate random-effect meta-analyses were performed to calculate pooled diagnostic yield estimates.ResultsFrom 1483 citations, 54 were selected for full-text review, and nine were included. Based on six studies including 256 children with microbiologically confirmed TB, the diagnostic yield of NAAT on one NPA ranged from 31 to 60% (summary estimate 44%, 95% CI 36–51%). From seven studies including 242 children with confirmed TB, the diagnostic yield of culture was 17–88% (summary estimate 58%, 95% CI 42–73%). Testing a second NPA increased the yield by 8–19% for NAAT and 4–35% for culture. NPA collection procedures varied between studies, although most children had NPA successfully obtained (96–100%), with a low rate of indeterminate results (< 5%). Data on NPA acceptability and specifically for children under 5 years were limited.ConclusionsNPA is a suitable and feasible specimen for diagnosing pediatric TB. The high rates of successful collection across different levels of healthcare improve access to microbiological testing, supporting its inclusion in diagnostic algorithms for TB, especially if sampling is repeated. Future research into the acceptability of NPA and how to standardize collection to optimize diagnostic yield is needed.