Mycobacterium Tuberculosis In Sputum Research Articles

Xpert MTB/RIF Ultra versus mycobacterial growth indicator tube liquid culture for detection of Mycobacterium tuberculosis in symptomatic adults: a diagnostic accuracy study

Xpert MTB/RIF Ultra (Ultra) is an automated molecular test for the detection of Mycobacterium tuberculosis in sputum. We compared the sensitivity of Ultra to that of mycobacterial growth indicator tube (MGIT) liquid culture, considered the most sensitive assay in routine clinical use. In this prospective, multicentre, cross-sectional diagnostic accuracy study, we used a non-inferiority design to assess whether the sensitivity of a single Ultra test was non-inferior to that of a single liquid culture for detection of Mtuberculosis in sputum. We enrolled adults (age ≥18 years) with pulmonary tuberculosis symptoms in 11 countries and each adult provided three sputum specimens with a minimum volume of 2 mL over 2 days. Ultra was done directly on sputum 1, and Ultra and MGIT liquid culture were done on resuspended pellet from sputum 2. Results of MGIT and solid media cultures done on sputum 3 were considered the reference standard. The pre-defined non-inferiority margin was 5·0%. Between Feb 18, 2016, and Dec 4, 2019, we enrolled 2906 participants. 2600 (89%) participants were analysed, including 639 (25%) of 2600 who were positive for tuberculosis by the reference standard. Of the 2357 included in the non-inferiority analysis, 877 (37%) were HIV-positive and 984 (42%) were female. Sensitivity of Ultra performed directly on sputum 1 was non-inferior to that of sputum 2 MGIT culture (MGIT 91·1% vs Ultra 91·9%; difference -0·8 percentage points; 95% CI -2·8 to 1·1). Sensitivity of Ultra performed on sputum 2 pellet was also non-inferior to that of sputum 2 MGIT (MGIT 91·1% vs Ultra 91·9%; difference -0·8 percentage points; -2·7 to 1·0). For the detection of M tuberculosis in sputum from adults with respiratory symptoms, there was no difference in sensitivity of a single Ultra test to that of a single MGIT culture. Highly sensitive, rapid molecular approaches for M tuberculosis detection, combined with advances in genotypic methods for drug resistance detection, have potential to replace culture. US National Institute of Allergy and Infectious Diseases.

Prevalence of subclinical pulmonary tuberculosis in adults in community settings: an individual participant data meta-analysis

Subclinical pulmonary tuberculosis, which presents without recognisable symptoms, is frequently detected in community screening. However, the disease category is poorly clinically defined. We explored the prevalence of subclinical pulmonary tuberculosis according to different case definitions. We did a one-stage individual participant data meta-analysis of nationally representative surveys that were conducted in countries with high incidence of tuberculosis between 2007 and 2020, that reported the prevalence of pulmonary tuberculosis based on chest x-ray and symptom screening in participants aged 15 years and older. Screening and diagnostic criteria were standardised across the surveys, and tuberculosis was defined by positive Mycobacterium tuberculosis sputum culture. We estimated proportions of subclinical tuberculosis for three case definitions: no persistent cough (ie, duration ≥2 weeks), no cough at all, and no symptoms (ie, absence of cough, fever, chest pain, night sweats, and weight loss), both unadjusted and adjusted for false-negative chest x-rays and uninterpretable culture results. We identified 34 surveys, of which 31 were eligible. Individual participant data were obtained and included for 12 surveys (620 682 participants) across eight countries in Africa and four in Asia. Data on 602 863 participants were analysed, of whom 1944 had tuberculosis. The unadjusted proportion of subclinical tuberculosis was 59·1% (n=1149/1944; 95% CI 55·8-62·3) for no persistent cough and 39·8% (773/1944; 36·6-43·0) for no cough of any duration. The adjusted proportions were 82·8% (95% CI 78·6-86·6) for no persistent cough and 62·5% (56·6-68·7) for no cough at all. In a subset of four surveys, the proportion of participants with tuberculosis but without any symptoms was 20·3% (n=111/547; 95% CI 15·5-25·1) before adjustment and 27·7% (95% CI 21·0-36·4) after adjustment. Tuberculosis without cough, irrespective of its duration, was more frequent among women (no persistent cough: adjusted odds ratio 0·79, 95% CI 0·63-0·97; no cough: adjusted odds ratio 0·76, 95% CI 0·62-0·93). Among participants with tuberculosis, 29·1% (95% CI 25·2-33·3) of those without persistent cough and 23·1% (18·8-27·4) of those without any cough had positive smear examinations. The majority of people in the community who have pulmonary tuberculosis do not report cough, a quarter report no tuberculosis-suggestive symptoms at all, and a quarter of those not reporting any cough have positive sputum smears, suggesting infectiousness. In high-incidence settings, subclinical tuberculosis could contribute considerably to the tuberculosis burden and to Mycobacterium tuberculosis transmission. Mr Willem Bakhuys Roozeboom Foundation.

Modelling of tuberculosis dynamics incorporating indirect transmission of contaminated environment and infectivity of smear-negative individuals: A case study for Xinjiang, China

Xinjiang has been one of the high incidence areas of pulmonary tuberculosis (PTB) in China. Besides being infected by direct contacting with active PTB individuals (direct infection), the susceptible would be infected because of the exposure to the environment contaminated by Mycobacterium tuberculosis (indirect infection). Active PTB individuals include not only the smear-positive PTB (PTB+) but also the smear-negative PTB (PTB–) who are infectious due to their ability to release tiny Mycobacterium tuberculosis particles even in the absence of visible Mycobacterium tuberculosis in sputum. By taking account of direct/indirect infection and the difference between PTB+ and PTB– individuals in transmission capability, a periodic dynamical PTB transmission model is proposed. The model is fitted to the newly monthly PTB+ and PTB– cases in Xinjiang from 2008 to 2017 by Markov Chain Monte Carlo algorithm. Moreover, global sensitivity analysis is constructed to address the uncertainty of some key parameters by using Latin hypercube sampling and partial rank correlation coefficient methods. Basic reproduction number R0 for PTB transmission in Xinjiang is estimated to be 2.447 (95% CrI:(1.203, 3.844)), indicating that PTB has been prevalent in Xinjiang over the study period. Our results suggest that reducing the direct/indirect transmission rates, early screening, isolating and treating the latent, PTB+ and PTB– individuals, and enhancing the clearance of Mycobacterium tuberculosis in the environment could more effectively control PTB transmission in Xinjiang. The model fits the reported PTB data well and achieves acceptable prediction accuracy. We believe that our model can provide heuristic support for controlling PTB transmission in Xinjiang.

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.

Effects of smoking on the severity and transmission of pulmonary tuberculosis: A hospital-based case control study.

There is a high incidence of both smoking and tuberculosis (TB) in China. This study examined the risk factors for severe pulmonary TB (PTB) and positive tubercle bacilli in sputum. We conducted a retrospective case-control study in a tertiary hospital from January 2017 to December 2018 (n = 917). The clinical and biological characteristics of patients were collected, and univariable and multivariable logistic regression analyses were performed to assess the factors associated with smoking in terms of the severity and transmission of PTB. Positive tubercle bacilli in sputum and severe PTB were much higher in smoking patients. Together with nutrition status, heavy smoking exhibited a 284% greater risk in severe PTB. Positive tubercle bacilli in sputum was significantly associated with hypoproteinemia and smoking regardless of the status, duration, and degree. Because cigarette smoking was strongly and inversely associated with hypoproteinemia, we conclude that smoking plays a critical role in the severity and transmission of PTB. Smoking cessation interventions should be employed to prevent severe PTB and decrease the transmission of PTB.

Quantification of multidrug-resistant M. tuberculosis bacilli in sputum during the first 8 weeks of treatment.

SETTING: Mulago Hospital, Kampala, Uganda.OBJECTIVE: To quantify Mycobacterium tuberculosis in sputum during the first 8 weeks of pulmonary multidrug-resistant TB (MDR-TB) treatment.DESIGN: We enrolled consecutive adults with pulmonary MDR-TB treated according to national guidelines. We collected overnight sputum samples before treatment and weekly. Sputum samples were cultured on Middlebrook 7H11S agar to measure colony-forming units per mL (cfu/mL) and in MGIT™ 960™ media to measure time to detection (TTD). Linear mixed-effects regression was used to estimate the relational change in log10 cfu/mL and TTD.RESULTS: Twelve adults (median age: 27 years) were enrolled. Half were women, and two-thirds were HIV-positive. At baseline, median log10 cfu/mL was 5.1, decreasing by 0.29 log10 cfu/mL/week. The median TTD was 116.5 h, increasing in TTD by 36.97 h/week. The weekly change was greater in the first 2 weeks (-1.04 log10 cfu/mL/week and 120.02 h/week) than in the remaining 6 weeks (-0.17 log10 cfu/mL/week and 26.11 h/week).CONCLUSION: Serial quantitative culture measures indicate a slow, uneven rate of decline in sputum M. tuberculosis over 8 weeks of standardized pulmonary MDR-TB treatment.

Rapid electrochemical detection of Mycobacterium tuberculosis in sputum by measuring Ag85 activity with disposable carbon sensors

An electrochemical assay for the detection of the enzymatic activity of the antigen 85 (Ag85) tuberculosis (TB) biomarker was developed and evaluated for the qualitative detection of Mycobacterium tuberculosis in decontaminated sputum. For this purpose, the electroactive properties of both synthetic p-aminophenyl-6-O-octanoyl-3-d-glucopyranoside (p-APOG) substrate and p-aminophenyl-6-3-d-glucopyranoside (p-APG) product released after the removal of the octanoyl fatty acid by the Ag85 were investigated with disposable carbon screen-printed electrodes by cyclic voltammetry. Since specific anodic responses were obtained for the p-APOG substrate and the p-APG product, the intensity of the oxidation peak of the p-APG (E = + 0.35 V vs. Ag/AgCl) was selected as the analytical response for the detection of the Ag85 acyltransferase activity. Once the proof of concept of the Ag85 electrochemical assay was validated with a commercially-available Ag85B protein, its specificity was further assessed by analyzing pure cultures of various bacteria including tuberculous and non-tuberculous mycobacteria as well as different species found in patients' sputum. Finally, with a specificity of 78% and a sensitivity of 89%, the method was successfully compared to microscopy and culture routine tests for TB testing in 36 frozen fluidized and decontaminated sputum. This suggests that owing to its convenience, rapidity, low-cost and portability, the reported Ag85 electrochemical assay is a promising tool to screen patients for TB.

Association of IL – 23 R rs 7518660 Gene Polymorphism with Susceptibility and Disease Severity of Pulmonary Tuberculosis

Pulmonary Tuberculosis (TB) is a global health problem. Of all people infected with Mycobacterium tuberculosis only a small proportion develops into TB. IL 23 is the key cytokine in the pathogenesis of TB infection. This study aims to determine the association of IL-23 R rs 7518660 gene polymorphism with susceptibility and disease severity of Pulmonary TB. A case control study involved 105 people consisting of 31 drug sensitive pulmonary TB patients, 40 patients with drug-resistant pulmonary TB and 34 healthy subjects as a control. IL-23 R rs 7518660 gene polymorphism G allele increases susceptibility to both TB drug-sensitive and drug-resistant. G and A allele, AA and AG genotypes indicates (p value >0.05) in correlation with disease severity based on lesion in chest x-ray and high load of Mycobacterium tuberculosis in sputum. There was a significant relationship between allele A and susceptibility to pulmonary TB with an odds ratio of 0.231. It showed that patients with A alleles (AG and AA genotypes) were at risk of developing TB by 1/0.231 = 4.33 times lower than patients with GG genotypes. Meanwhile, the relationship of the G allele with susceptibility to pulmonary TB obtained (p value <0.05) and an odds ratio value of 0.127 indicating that patients with G alleles (GG and AG genotypes) were at risk of developing TB of 1/0.127 = 7.87 times higher than in patients with the AA genotype. Conclusion: We found significant correlation between IL-23 R rs 7518660 gene polymorphism G allele with susceptibility to pulmonary TB, but the result was not significant with disease severity.

MOLECULAR DETECTION OF INSERTION SEQUENCE 6110 OF MYCOBACTERIUM TUBERCULOSIS IN PATIENTS WITH PULMONARY TUBERCULOSIS AND TUBERCULOUS PLEURITIS IN ANBAR GOVERNORATE, WEST OF IRAQ

Accurate identification and early diagnosis of tuberculosis especially latent and active infection is the key to prevention of the disease. This study was conducted to detect the accuracy of conventional and real time polymerase chain reaction in the diagnosis of pulmonary tuberculosis caused by Mycobacterium tuberculosis in sputum and blood samples using 123 bp gene of repetitive insertion sequence 6110 (IS6110) of bacterial genome and tuberculous pleuritis using pleural effusion. Sixty five patients who have clinical suspicion of pulmonary tuberculosis, fifty patients with suspicion of TB pleurisy and twenty patients with non-tuberculous mycobacterial pulmonary disease as control were studied during the period from April, 2012 to December, 2015. They were admitted to Department of Internal Medicine in Ramadi Teaching Hospital and Clinical Private. EDTA-Peripheral blood and sputum samples had been taken from patient with pulmonary tuberculosis. Also, pleural effusion was obtained from patients with tuberculous pleuritis. They were subjected to DNA extraction, amplification of the target DNA by conventional PCR and qualitative real time PCR. Of the 65 sputum samples from patients with pulmonary tuberculosis, 37 (56.9%) were AFB smear-positive. Of these, 32 (86.5%) and ?? (94.6%) were positive for PCR and RTPCR respectively. Further, out of 28 negative smear, 7 (25%) and 8 (28.6%) smears were PCR and RTPCR positive respectively. Further, in peripheral blood based PCR study, out of 37 patients whose sputum were positive by AFB smear, only 25 (67.6%) of them were positive for PCR. Peripheral blood based PCR and RT-PCR essays were negative for all AFB smear negative cases. In suspected tuberculous pleuritis, 8 (16%), 9 (18%) patients reveal positive result for both of conventional and real time PCR respectively. Also, all twenty spontaneous sputum samples (controls)were negative for PCR. The study concluded that PCR provides a sensitive and specific means for laboratory diagnosis of pulmonary tuberculosis and qualitative real time PCR is a more confirmatory test but the molecular diagnosis of the tuberculosis should be based on the combined analysis of baciloscopia, clinical manifestations and therapeutic proves. Further, it was concluded that PCR and RT-PCR based- peripheral blood leukocytes is of little value for specific diagnosis of pulmonary tuberculosis. Regarding tuberculous pleuritis, PCR test revealed low sensitivity and high specificity using pleural fluid. The sensitivity was higher in cases in which the bacillary load was high-in acid fast bacilli-positive samples.

Elevated Plasma Matrix Metalloproteinase 8 Associates With Sputum Culture Positivity in Pulmonary Tuberculosis.

Current methods for tuberculosis treatment monitoring are suboptimal. We evaluated plasma matrix metalloproteinase (MMP) and procollagen III N-terminal propeptide concentrations before and during tuberculosis treatment as biomarkers. Plasma MMP-1, MMP-8, and MMP-10 concentrations significantly decreased during treatment. Plasma MMP-8 was increased in sputum Mycobacterium tuberculosis culture–positive relative to culture-negative participants, before (median, 4993 pg/mL [interquartile range, 2542–9188] vs 698 [218–4060] pg/mL, respectively; P = .004) and after (3650 [1214–3888] vs 720 [551–1321] pg/mL; P = .008) 6 months of tuberculosis treatment. Consequently, plasma MMP-8 is a potential biomarker to enhance tuberculosis treatment monitoring and screen for possible culture positivity.

Portable immunosensor directly and rapidly detects Mycobacterium tuberculosis in sputum.

Tuberculosis (TB) remains a public health problem that cannot be ignored. The portable and efficient detection of Mycobacterium tuberculosis (MTB) is important for the effective control of this disease. However, current detection techniques do not meet the requirements for MTB detection in the actual environment and often require cumbersome detection steps that are time consuming and inflexible. In this study, a portable immunosensor to detect MTB in sputum was prepared and then subjected to interface characterizations, such as scanning electron microscopy, hydrophilic angle test, and fluorescence characterization. The source and gate voltage of the device were optimized and tested using a non-contact photoresponse. The results showed that the sensitivity of the sensor to luminance increases with the decrease in source voltage. The gate voltage can substantially improve the response of the immunosensor to the normalized current of protein and amplify the signal at least 1.6 times. The optimal voltage detection conditions of source voltage (0.3 V) and gate voltage (0.1 V) were also determined. Several common proteins present in simulated saliva were used for anti-interference tests, and the sensor exhibited good specificity. Finally, the dilution gradient of an actual TB sputum sample was optimized. In the absence of preconditioning, a double-blind experiment was used to distinguish between the sputum from patients with TB and healthy individuals to shorten the TB detection time to a few minutes. Compared with the hospital's conventional detection method using cultures, the proposed method can complete the detection in a shorter time. This study provides a new strategy for the portable diagnosis of TB.

Prevalence of Mycobacterium tuberculosis in Sputum and Reported Symptoms Among Clinic Attendees Compared With a Community Survey in Rural South Africa.

BackgroundTuberculosis (TB) case finding efforts typically target symptomatic people attending health facilities. We compared the prevalence of Mycobacterium tuberculosis (Mtb) sputum culture-positivity among adult clinic attendees in rural South Africa with a concurrent, community-based estimate from the surrounding demographic surveillance area (DSA).MethodsClinic: Randomly selected adults (≥18 years) attending 2 primary healthcare clinics were interviewed and requested to give sputum for mycobacterial culture. Human immunodeficiency virus (HIV) and antiretroviral therapy (ART) status were based on self-report and record review. Community: All adult (≥15 years) DSA residents were invited to a mobile clinic for health screening, including serological HIV testing; those with ≥1 TB symptom (cough, weight loss, night sweats, fever) or abnormal chest radiograph were asked for sputum.ResultsClinic: 2055 patients were enrolled (76.9% female; median age, 36 years); 1479 (72.0%) were classified HIV-positive (98.9% on ART) and 131 (6.4%) reported ≥1 TB symptom. Of 20/2055 (1.0% [95% CI, .6–1.5]) with Mtb culture-positive sputum, 14 (70%) reported no symptoms. Community: 10 320 residents were enrolled (68.3% female; median age, 38 years); 3105 (30.3%) tested HIV-positive (87.4% on ART) and 1091 (10.6%) reported ≥1 TB symptom. Of 58/10 320 (0.6% [95% CI, .4–.7]) with Mtb culture-positive sputum, 45 (77.6%) reported no symptoms. In both surveys, sputum culture positivity was associated with male sex and reporting >1 TB symptom.ConclusionsIn both clinic and community settings, most participants with Mtb culture-positive sputum were asymptomatic. TB screening based only on symptoms will miss many people with active disease in both settings.

Aptasensor for the Detection of Mycobacterium tuberculosis in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker.

A portable electrochemical aptamer-antibody based sandwich biosensor has been designed and successfully developed using an aptamer bioreceptor immobilized onto a screen-printed electrode surface for Mycobacterium tuberculosis (M. tuberculosis) detection in clinical sputum samples. In the sensing strategy, a CFP10-ESAT6 binding aptamer was immobilized onto a graphene/polyaniline (GP/PANI)-modified gold working electrode by covalent binding via glutaraldehyde linkage. Upon interaction with the CFP10-ESAT6 antigen target, the aptamer will capture the target where the nano-labelled Fe3O4/Au MNPs conjugated antibody is used to complete the sandwich format and enhance the signal produced from the aptamer–antigen interaction. Using this strategy, the detection of CFP10-ESAT6 antigen was conducted in the concentration range of 5 to 500 ng/mL. From the analysis, the detection limit was found to be 1.5 ng/mL, thereby demonstrating the efficiency of the aptamer as a bioreceptor. The specificity study was carried out using bovine serum albumin (BSA), MPT64, and human serum, and the result demonstrated good specificity that is 7% higher than the antibody–antigen interaction reported in a previous study. The fabricated aptasensor for M. tuberculosis analysis shows good reproducibility with an relative standard deviation (RSD) of 2.5%. Further analysis of M. tuberculosis in sputum samples have shown good correlation with the culture method with 100% specificity and sensitivity, thus making the aptasensor a promising candidate for M. tuberculosis detection considering its high specificity and sensitivity with clinical samples.

Pathological and DNA-based Detection of Tracheobronchial Tuberculosis in China.

To compare the DNA-based sputum evaluation with histopathology for the diagnosis of tracheo-bronchial tuberculosis (TBTB). Case series. Place and Duration of Study: Tianjin Haihe Hospital, Tianjin Institute of Respiratory Diseases, Tianjin, China, from January to June 2017. TBTB patients, who underwent bronchoscopy during the study period, were included. Their bronchoscopy presentations, histopathology, and induced sputum deoxyribonucleic acid (DNA) and culture results were analysed. Results were expressed as frequency percentage. There were 110 subjects. Most patients were young females with coughing. The main TBTB subtype was fibrostenotic, while the most common findings were single level lesions and lobar bronchi. The diagnostic rates were 40.43%, 67.86%, and 68.54% for DNA-based analysis of induced sputum, histopathology, and induced sputum culture, respectively. Only in the edematous-hyperemic subtype was the positivity rate of DNA-based analysis of induced sputum higher than in histopathology. In the fibrostenotic subtype, the histopathologic results were superior for diagnosis. A combination of induced sputum and biopsy using DNA-based methods is a superior and exact method to diagnose TBTB. DNA-based analysis of induced sputum for mycobacterium tuberculosis can be used for preliminary impressions. Key Words: Tracheo-bronchial tuberculosis, Mycobacterium tuberculosis, Pathology, Multiplex polymerase chain reaction, Sputum.

ИЗУЧЕНИЕ РЕЗИСТЕНТНОСТИ К ПРОТИВОТУБЕРКУЛЕЗНЫМ СРЕДСТВАМ У БОЛЬНЫХ, НАХОДЯЩИХСЯ НА СТАЦИОНАРНОМ ЛЕЧЕНИИ В ОТДЕЛЕНИИ РЕЗИСТЕТНЫХ ФОРМ ТУБЕРКУЛЕЗА ЛЕГКИХ

A comparative study of resistance to anti-tuberculosis drugs in patients with newly diagnosed pulmonary tuberculosis and in patients with chronic forms of tuberculosis who were treated in an anti-tuberculosis hospital was carried out. We examined 142 patients treated in the department of resistant forms of tuberculosis. A complete bacteriological study of sputum for Mycobacterium tuberculosis was carried out with the determination of sensitivity to anti-tuberculosis drugs during the cultivation of microorganisms on a liquid nutrient medium in a system with automated registration of bacterial growth and on a dense nutrient medium of Lowenstein-Jensen using the method of absolute concentrations. Methods of PCR analysis were also used. According to the anamnesis, two groups of patients were formed: 82 patients with newly diagnosed tuberculosis and 60 patients with a chronic course of the disease or with a relapse of the disease. According to the age structure, patients under the age of 50 prevailed in both groups. At the same time, in the first group there was a significant number of patients aged 18-29 years - 20.7%. In the second group, the number of patients aged 18-29 years was significantly less - 5%. Despite the fact that all patients were assigned to the category of multidrug-resistant pathogen and were treated in the unit of resistant forms of tuberculosis, it was important to assess the level of Mycobacterium tuberculosis resistance separately for patients in each of the selected groups. The largest differences between groups were found in the assessment of second-line drug resistance. Patients of the second group showed high resistance to fluoroquinolones - 30% and amikacin - 41.6%. In patients with newly diagnosed pulmonary tuberculosis (first group), resistance to fluoroquinolones was 9.7%, and to amikacin - 17.1%. Capreomycin was the most effective in both groups. The resistance of the causative agent of tuberculosis in patients of the first group was 4.9%, in the second - 11.6%.

Diagnostic accuracy of a novel point-of-care urine lipoarabinomannan assay for the detection of tuberculosis among adult outpatients in Zambia: a prospective cross-sectional study.

BackgroundA novel, rapid, point-of-care urine-based lipoarabinomannan assay (Fujifilm SILVAMP TB LAM (“FujiLAM”)) has previously demonstrated substantially higher sensitivity for tuberculosis (TB) compared with the commercially available Determine TB LAM assay using biobanked specimens. However, FujiLAM has not been prospectively evaluated using fresh urine specimens. Therefore, we determined the diagnostic accuracy of FujiLAM among HIV-positive and HIV-negative outpatients with presumptive TB in Zambia.MethodsAdult (≥18 years old) presumptive TB patients presenting to two outpatient public health facilities in Lusaka were included. All patients submitted sputa samples for smear microscopy, Xpert MTB/RIF and mycobacterial culture, and urine samples for the FujiLAM assay. Microbiologically confirmed TB was defined by the detection of Mycobacterium tuberculosis in sputum using culture; this served as the reference standard to assess the diagnostic accuracy of FujiLAM.Results151 adults with paired sputum microbiological tests and urine FujiLAM results were included; 45% were HIV-positive. Overall, 34 out of 151 (23%) patients had culture-confirmed pulmonary TB. The overall sensitivity and specificity of FujiLAM was 77% (95% CI 59–89%) and 92% (95% CI 86–96%), respectively. FujiLAM's sensitivity among HIV-positive patients was 75% (95% CI 43–95%) compared with 75% (95% CI 51–91%) among HIV-negative patients. The sensitivity of FujiLAM in patients with smear-positive, confirmed pulmonary TB was 87% (95% CI 60–98%) compared with 68% (95% CI 43–87%) among patients with smear-negative, confirmed pulmonary TB.ConclusionsFujiLAM demonstrated high sensitivity for the detection of TB among both HIV-positive and HIV-negative adults, and also demonstrated good specificity despite the lack of systematic extrapulmonary sampling to inform a comprehensive microbiological reference standard.

Tuberculosis and COVID-19 co-infection: A case series from Iran

Aims and objectives: Because the world is mobilized with all its might to fight the COVID-19 epidemic, control of common but deadly diseases such as tuberculosis may be neglected. With a conservative estimate and modeling working, we can show that if too much attention to the COVID-19 continuing it can leads to a 25% reduction in global TB diagnosis by 3 months and TB mortality will increase by 13% and this brings us back to the level of TB mortality we had 5 years ago. Between 2020 and 2025, 1.4 million TB deaths could be recorded as a direct consequence of COVID-19. Therefore, ensuring the continuation of basic services and operations to deal with long-standing health problems like tuberculosis or other similar diseases to protect the lives of people is very important. Methods: This study is a case series has been performed in three hospital centers in Tehran where respiratory and infectious patients were treated. We report on eleven cases of patients who were either unaware of their tuberculosis or of their COVID-19 and after visiting a medical center, they found out that they have both of them. Results: Of the eleven patients in this study, seven were male and four were female, with a mean age of 56.6 years (minimum age 27 years and maximum age 91 years). Five patients were previously diagnosed with tuberculosis and six patients were initially admitted with a diagnosis of COVID-19. Seven were Afghans and four were Iranians. Ten patients had respiratory tuberculosis and one had both pulmonary and spinal tuberculosis. Fever, productive cough, and hemoptysis were present in all of them. After clinical suspicion, spiral lung CT-scan, COVID-19 RT-PCR assay, smear and culture of sputum for mycobacterium tuberculosis, were performed and finally simultaneous tuberculosis and COVID-19 co-infection was diagnosed. Two patients died in the first week of treatment after being admitted to the intensive care unit. However, both patients also had diabetes mellitus and hypertension. Nine other cases were transferred to the tuberculosis unit after 7 to 10 days of treatment in the COVID-19 ward. Conclusions: Tuberculosis and COVID-19 are both common infections of the respiratory system and have common signs and symptoms and these similarities may cause overlapping the two diseases. While experience with COVID-19 infection in TB patients is still limited, it is predicted that people with TB and COVID-19 may have poorer treatment outcomes, especially if TB treatment is postponed.

Comparison of staining techniques– Ziehl Neelsen stain, Gabbet’s Stain, Fluorochrome stain for detection of mycobacterium tuberculosis in sputum

In developing countries, Ziehl-Neelsen (Z-N) sputum smear microscopy is widely used.The present prospective study was under taken to see the efficacy of Ziehl-Neelson method versus Gabbett and Fluorescent staining in the detection of mycobacteria in sputum sample. 80 sputum samples were collected from suspected tuberculosis patients attending RNTCP OPD at, Nerul, Navi Mumbai. Triplicate smears were prepared from all these samples and stained by the Z-N, Gabbet’s, and Fluorescent stain. Number of smears positive for Acid Fast Bacilli by, Z-N method 13(16%), Gabbett’s method 7(9%), Fluorescent method 26(33%). Combination of two staining methods yielded following results: Z- N + Fluorescent26 (33%) and Z- N + Gabbett 13(16%). The conclusion from this study is that when smear examination by Z-N is done, there are possibilities of false negative results which can be reduced by using Fluorescent staining. The rate of positivity can be improved by combination of Z-N + Fluorescent staining. So, it may be concluded that combination of ZN and Fluorescent Microscopy may be considered for diagnosis of pulmonary tuberculosis in any diagnostic set up along with culture which is the gold standard. Keywords: Fluorescent, Gabbett’s, Mycobacterium tuberculosis, Revised National Tuberculosis Control Programme (RNTCP), Zehil-Neelson (Z-N).

Pulmonary tuberculosis at Patan Hospital, Nepal: One year audit

BackgroundGlobally, Tuberculosis (TB) is one of the top 10 causes of death. In Nepal, poverty and malnutrition aggravate the burden of TB. To identify Mycobacterium tuberculosis sputum is the best sample to identify the bacterium which is helpful for diagnosis. The aim of this study is to identify the situation, burden and challenges of pulmonary tuberculosis in low-middle income country like Nepal. MethodA retrospective-audit with reliable-secondary-data of one year was collected (n = 4131). Descriptive-analysis was performed using frequency, percentage and analytical using chi-square-test. Level of significance was set at p < 0.05. Ethical Approval was obtained from IRC-PAHS. ResultThe prevalence of notified/suspected cases was highest among the patients having >60 years of age 1344(32.54%) and least among the patients with ≤15 years of age 239(5.79%). The male had 1.67 times more smear-positive pulmonary TB cases. Among the AFB-positive cases, smear 3+ was seen in most of the cases 69(38.54%) followed by smear 1+ and smear 2+ in 56(31.28%) and 54(30.16%) respectively. ConclusionThe prevalence of smear-positive pulmonary TB case is higher in male. Smear 3+ is seen in most of the followed by smear 1+ and smear 2+. The Burden of Pulmonary TB is more among adult and old-age-people and its control is a challenge for developing and low-middle income countries like Nepal.

GeneXpert MTB/RIF and Mycobacterium tuberculosis Sputum Culture in Establishing the Diagnosis of Pulmonary Tuberculosis and Rifampicin Resistance in Suspected Childhood Pulmonary Tuberculosis in Soetomo Hospital

The diagnosis of childhood tuberculosis remains a challenge worldwide. The GeneXpert MTB/RIF test, a rapid Mycobacteria tuberculosis diagnostic tool, was recommended for use in children. No pediatric studies of GeneXpert MTB/RIF assessing pulmonary tuberculosis within a hospital setting has been done in Indonesia. We evaluated the performance of the GeneXpert MTB/RIF test compared with sputum culture on Lowenstein-Jensen (LJ) for the diagnosis of childhood pulmonary tuberculosis. This study was conducted in pediatric respirology inpatient and outpatient Dr. Soetomo Hospital, a tertiary care facility in Surabaya between June and August 2015 with a cross-sectional design. We consecutively enrolled 27 children aged 3 months to 14 years who had history of close contact with adult tuberculosis patients and showed symptoms of pulmonary tuberculosis. Sputum collection was performed by induced sputum and three examination methods were performed (microscopic, GeneXpert MTB/RIF and sputum culture) simultaneously followed by a drug sensitivity test for specimens detected with MTB growth. The GeneXpert MTB/RIF test had a sensitivity of 100% (95% CI 100-100) and a specificity of 95% (95% CI 85-100). The positive predictive value for diagnosing pulmonary TB was 89% (95% CI 68-100), the negative predictive value was 100% (95% CI 100-100) and positive likelihood ratio was 20 (95% CI 2.82-128). The GeneXpert MTB/RIF test on one sputum sample rapidly and correctly identified all children with culture-confirmed pulmonary tuberculosis with high specificity. Similar results were obtained between GeneXpert MTB/RIF and sputum culture based on age groups and clinical manifestations. Rifampicin resistance were both detected in GeneXpert MTB/RIF and MTB sensitivity test