Diagnosis Of Active Tuberculosis Research Articles

COMPARATIVE ASSESSMENT OF DIFFERENT DIAGNOSTIC TECHNIQUES IN THE IDENTIFICATION OF MYCOBACTERIUM TUBERCULOSIS IN SUSPECTED CASES OF TUBERCULOSIS

Statement of the Problem: Tuberculosis remains a serious public-health threat in developing countries though it has been eradicated in some advanced countries. This disease constitutes a significant threat to global health, being the second highest cause of morbidity and mortality resulting from infectious agents. Prompt diagnosis of active TB facilitates timely therapeutic intervention and minimizes community transmission. Aim: This study aimed at determining a ‘Point of Care’ diagnostic tool for pulmonary tuberculosis (PTB) by comparing the efficiency of four different PTB diagnostic tools for different age groups. Methodology: Zeihl Nelson (ZN) staining, culture, Gene xpert (GX) and Lipoarabinomanan (LAM) assay were employed in this study The culture method was used for confirmation. Sputum and urine samples were collected from each of 100 patients symptomatically diagnosed of PTB. Findings: Fifty-seven percent of the population was male while 43% were female. Mycobacterium tuberculosis was isolated from 9 (9%) of 100 patients. Similarly, GX detected Mycobacterium tuberculosis in 9 (9%) of the patients while the rate of detection using LAM was 10% and with ZN it was 7%. Gene xpert produced no true or false positive and negative result, LAM had one false positive result and ZN had two false negative results. The maximum time frame to generate result was 25 minutes for LAM, two hours for Gene xpert, eight weeks for culture and two days for ZN. Two positive isolates were observed at the same frequency for age group 21-30 and 31- 40 while age groups 1-10, 10-20, 41-50, 50-60 and above has 1 positive result each. Gene xpert had 98.11% sensitivity while LAM had 96.23% and ZN had 86.79%. The choice of ‘Point of Care’ diagnostic tool is of great concern to clinicians and the general public. Conclusion & Significance: This study identified LAM assay as suitable ‘Point of Care’ diagnostic and an add-on tool for PTB diagnosis because of its relatively high sensitivity and short maximum time frame to generate result compare to other three diagnostic techniques.
 
 

Mycobacterium tuberculosis latency-associated antigen Rv1733c SLP improves the accuracy of differential diagnosis of active tuberculosis and latent tuberculosis infection.

Background:Differential diagnosis of active tuberculosis (ATB) and latent tuberculosis infection (LTBI) has been a challenge for clinicians in high TB burden countries. The purpose of this study was to improve the accuracy of differential diagnosis of ATB and LTBI by using fluorescent immunospot (FluoroSpot) assay to detect specific Th1 cell immune responses. The novel mycobacterium tuberculosis (MTB) latency-associated antigens Rv1733c and synthetic long peptides derived from Rv1733c (Rv1733c SLP) were used based on virulence factors early secreting antigen target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10).Methods:Fifty-seven ATB cases, including 20 pathogen-confirmed ATB and 37 clinically diagnosed ATB, and 36 LTBI cases, were enrolled between January and December 2017. FluoroSpot assay was used to detect the interferon γ (IFN-γ) and interleukin 2 (IL-2) secreted by the specific T cells after being stimulated with MTB virulence factors ESAT-6 and CFP-10, MTB latency-associated antigens Rv1733c and Rv1733c SLP. The receiver operating characteristic (ROC) curve was used to define the best cutoff value of latency-associated antigens in the use of differentiating ATB and LTBI. The sensitivity, specificity, predictive value, and likelihood ratio of ESAT-6 and CFP-10-FluoroSpot combined with latency-associated antigen in the differential diagnosis of ATB and LTBI were also calculated.Results:Following the stimulation with Rv1733c and Rv1733c SLP, the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP had the largest area under the ROC curve, which was 0.766. With a cutoff value of 1 (spot-forming cells [SFCs]/2.5 × 105 peripheral blood mononuclear cells) for frequency, the sensitivity and specificity of distinguishing ATB from LTBI were 72.2% and 73.7%, respectively. ESAT-6 and CFP-10-FluoroSpot detected the frequency and proportion of single IFN-γ-secreting T cells; the sensitivity and specificity of distinguishing ATB from LTBI were 82.5% and 66.7%, respectively. Combined with the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP on the basis of ESAT-6 and CFP-10-FluoroSpot, the sensitivity and specificity increased to 84.2% and 83.3%, respectively.Conclusion:Rv1733c SLP, combined with ESAT-6 and CFP-10, might be used as a candidate antigen for T cell-based tuberculosis diagnostic tests to differentiate ATB from LTBI.

Sustaining Togetherness; Family Engagement in Tuberculosis Prevention in Children Living in Tuberculosis Households in Indonesia

BACKGROUND: Preventing tuberculosis (TB) in children needs the family to be engaged; however, limited studies about how the family engages in TB prevention. AIM: This aimed of study was to develop the process of family engagement in TB prevention for children with household contact of TB patient. METHODS: Guided by Straussian grounded theory, we developed a substantive theory from 14 primary caregivers in West Java-Indonesia as the key participants of families whose child ≤5 years old received TB screening and preventive therapy, and have had an adult in the household with active TB diagnosis. Data were collected by in-depth interviews and analyzed by coding procedures. RESULTS: Sustaining togetherness aimed to maintain the children’s health, which underpinned three main stages that consisted of “moving from fear to realization,” “making the family’s collective decision,” and “holding on together.” CONCLUSION: Implications for nursing practice in providing optimum care of preventing TB among children through engagement of family.

Clinical Features and Risk Factors for Active Tuberculosis in Takayasu Arteritis: A Single-Center Case-Control Study.

BackgroundsTakayasu arteritis (TAK) is a chronic, granulomatous vasculitis correlated with tuberculosis (TB). The two diseases share similar pathological characteristics and clinical manifestations which increase the difficulty to diagnose. Active tuberculosis (ATB) has implications for treatment strategies in TAK patients. Therefore, the investigation of clinical features and potential risk factors of ATB in TAK patients is vital.MethodsThe study reviewed hospitalized patients diagnosed with TAK in our hospital from 2008, to 2021. TAK patients with ATB were enrolled as the case group. The control group was randomly selected in a 3:1 ratio. The clinical characteristics of TAK patients with and without ATB were compared. Multivariate logistic regression analysis was performed to determine risk factors for ATB in TAK patients.ResultsWe reviewed 1,789 patients and ultimately identified 30 (1.7%) ATB cases. TAK patients with ATB were more prone to develop symptoms including fever (p=0.001), fatigue (p=0.003), cough (p=0.037), expectoration (p<0.001), weight loss (p=0.003), and night sweating (p<0.001). Increased level of hypersensitive C reactive protein (hsCRP, p=0.001), decreased level of albumin (p=0.031), and higher positive rate of T-SPOT.TB test (p<0.001) were observed in the case group. Multivariate logistic regression analysis revealed that hsCRP >8 mg/L (OR 9.108; 95% CI, 1.096–75.711; p=0.041) and positive T-SPOT.TB result (OR 68.669; 95% CI, 7.291–646.738; p<0.001) were risk factors for ATB in TAK patients. The proportion of patients undergoing subsequent surgery for Takayasu arteritis was lower in patients with ATB (p<0.001).ConclusionOur study suggested that the diagnosis of ATB should be considered when TAK patients experienced symptoms including fever, fatigue, weight loss, etc. hsCRP >8 mg/L and positive T-SPOT.TB result were identified as independent risk factors for ATB in TAK patients.

Gaps in the tuberculosis preventive therapy care cascade in children in contact with TB

ABSTRACT Background Young children (<5 years) and children living with HIV in contact with an adult with tuberculosis (TB) should receive TB preventive therapy (TPT), but uptake is low. Aims To determine gaps in the uptake of and adherence to TPT in child TB contacts under routine primary care clinic conditions. Methods A cohort of child TB contacts (age <5 years or living with HIV <15 years) was followed at a primary care clinic in Johannesburg, South Africa. Results Of 170 child contacts with 119 adult TB cases, only 45% (77/170) visited the clinic for TPT eligibility screening, two of whom had already initiated TPT at another clinic. Of the 75 other children, 18/75 (24%) commenced TB treatment and 56/75 (75%) started TPT. Health-care workers followed the guidelines, with 96% (64/67) of children screened for symptoms of TB and 97% (36/37) of those symptomatic assessed for TB, but microbiological testing was low (9/36, 25%) and none had microbiologically confirmed tuberculosis. Only half (24/46, 52%) of the children initiating TPT completed the 6-month course. Neither sociodemographic determinants (age, sex) nor clinical factors (HIV status, TB source, time to TPT initiation) was associated with non-adherence to TPT. Conclusion Most child contacts of an adult TB case do not visit the clinic, and half of those initiating TPT did not adhere to the full 6-month course. These programme failures result in missed opportunities for early diagnosis of active TB and prevention of progression to disease in young and vulnerable children.

Automated machine learning for endemic active tuberculosis prediction from multiplex serological data

Serological diagnosis of active tuberculosis (TB) is enhanced by detection of multiple antibodies due to variable immune responses among patients. Clinical interpretation of these complex datasets requires development of suitable algorithms, a time consuming and tedious undertaking addressed by the automated machine learning platform MILO (Machine Intelligence Learning Optimizer). MILO seamlessly integrates data processing, feature selection, model training, and model validation to simultaneously generate and evaluate thousands of models. These models were then further tested for generalizability on out-of-sample secondary and tertiary datasets. Out of 31 antigens evaluated, a 23-antigen model was the most robust on both the secondary dataset (TB vs healthy) and the tertiary dataset (TB vs COPD) with sensitivity of 90.5% and respective specificities of 100.0% and 74.6%. MILO represents a user-friendly, end-to-end solution for automated generation and deployment of optimized models, ideal for applications where rapid clinical implementation is critical such as emerging infectious diseases.

Association of Urinary Interferon Gamma Protein-10 Levels and Low Levels of Cluster of Differentiation 4 Serum in Patients with Tuberculosis-Human Immunodeficiency Virus Coinfection

BACKGROUND: Tuberculosis (TB) infection caused by Mycobacterium tuberculosis was the most frequent opportunistic infection in human immunodeficiency virus (HIV) patients that lead to increasing of morbidity and mortality. Very low cluster of differentiation 4 (CD4) levels causing problem in TB/HIV diagnostic because unspecific clinical manifestation. Urine interferon gamma protein-10 (IP10) examination is a method that is relatively easier and safer to do, so it can be used as an alternative method in establishing the diagnosis TB/HIV. AIM: The aim of this study was to investigate the association between urinary IP-10 and level of CD4 serum in patients with TB/HIV coinfection. MATERIALS AND METHODS: This study was conducted at the Department of Internal Medicine, Faculty of Medicine, Dr. M. Djamil Hospital, Padang, Indonesia, involved 30 patients with active TB/HIV coinfection whose urine and blood serum were collected to evaluate IP-10 and CD4 level. RESULTS: There is a significant relationship between urine IP10 levels as a biomarker for the diagnosis of active pulmonary TB in HIV patients and serum CD4 levels in TB-HIV coinfected patients with moderate correlation strength. CONCLUSION: Further research is needed with a larger sample size to see the effect of low serum CD4 levels on the sensitivity of urine IP10 diagnosis.

Cell population data in identifying active tuberculosis and community-acquired pneumonia.

ObjectiveLeukocyte morphological parameters known as CPD (cell population data) is detected by hematology analyzer UniCel DxH800 with VCS technology. This study aimed to investigate the diagnostic efficacy of morphological changes in CPD parameters in distinguishing active tuberculosis from community-acquired pneumonia.MethodsFrom October 2018 to February 2019, 88 patients with active tuberculosis, 78 patients with community-acquired pneumonia, and 89 healthy controls were enrolled in this study. CPD was obtained using Unicel DxH800 analyzer for all whole blood samples, one-way ANOVA (non-parametric) and area analysis under ROC curve were performed.ResultsThe neutrophil mean conductivity (NMC), monocyte mean volume (MMV), monocyte mean conductivity (MMC), lymphocyte percentage (LY%), and monocyte percentage (MO%) were significantly higher in the active tuberculosis group than in the community-acquired pneumonia group. The white blood cell (WBC) count and neutrophil percentage (NE%) were significantly lower in the active tuberculosis group than in the community-acquired pneumonia group. The analysis of the area under the ROC curve proved that WBC count, neutrophil percentage (NE%), lymphocyte percentage (LY%), and monocyte percentage (MO%) did not achieve a higher area under the curve (AUC: 0.63, 0.71, 0.62, and 0.7, respectively). However, the AUC of NMC, MMV, and MMC in the CPD parameters was 0.951, 0.877, 0.98, respectively, and the simultaneous measurement of the three parameters was 0.99. The sensitivity and specificity were 98.5% and 91.1%, respectively.ConclusionThe combined diagnosis of NMC, MMV, and MMC could assist the clinical diagnosis of active tuberculosis and community-acquired pneumonia.

Utility of interferon gamma/tumor necrosis factor alpha FluoroSpot assay in differentiation between active tuberculosis and latent tuberculosis infection: a pilot study

BackgroundThe differential diagnosis of active tuberculosis (ATB) and latent tuberculosis infection (LTBI) remains challenging in clinical practice. We aimed to evaluate the diagnostic accuracy of the IFN-γ/TNF-α FluoroSpot assay for differentiating ATB from LTBI.MethodsWe conducted a pilot study of case-control design, using the FluoroSpot assay to simultaneously detect IFN-γ and TNF-α secretion at the single-cell level. The frequencies of antigen-specific single TNF-α-, total TNF-α-, single IFN-γ-, total IFN-γ- and dual IFN-γ/TNF-α-secreting T cells were detected. The optimal cutoffs value of frequencies for differentiating ATB from LTBI were determined according to receiver operating characteristic curve analysis. The sensitivity, specificity, predictive values (PV) and likelihood ratios (LR) of the FluoroSpot assay were calculated.ResultsThirty patients diagnosed microbiologically with ATB, 36 healthcare workers with LTBI and 36 healthy controls were enrolled. After stimulated by ESAT-6 or CFP-10 peptides, the median frequencies of single TNF-α-, total TNF-α-, single IFN-γ-, total IFN-γ- and dual IFN-γ/TNF-α-secreting T cells in ATB patients were all significantly higher than those in LTBI and HC groups (P < 0.01). The frequencies of total IFN-γ-secreting T cells detected by FluoroSpot assay correlated significantly with those of T-SPOT.TB (r = 0.910 for ESAT-6, P < 0.001, r = 0.845 for CFP-10, P < 0.001). After stimulated by ESAT-6 peptides, with total TNF-α-secreting T cells frequencies at a cut off value of 21 iSFCs/250,000 PBMCs, the sensitivity, specificity, PLR, NLR, PPV, NPV of IFN-γ/TNF-α FluoroSpot assay in differentiating ATB from LTBI were 96.7% (95%CI, 82.8–99.9%), 94.3% (95%CI, 80.8–99.3%), 16.92 (95%CI, 4.40–65.08), 0.04 (95%CI, 0.01–0.24), 93.6% (95%CI,78.6–99.2%) and 97.1% (95%CI, 84.7–99.9%), respectively. With the frequencies of total TNF-α- and total IFN-γ-secreting T cells stimulated by ESAT-6 peptides combined, the specificity was increased to 97.1%, and the positive likelihood ratio to 31.5. The combination with CFP-10 might not improve the diagnostic accuracy of the ESAT-6 for differentiating ATB from LTBI.ConclusionsIFN-γ/TNF-α FluoroSpot assay might have potential to help differentiate ATB from LTBI, but the findings need to be further verified by cross-sectional or prospective cohort studies.

German S3-Guideline on the treatment of Psoriasis vulgaris, adapted from EuroGuiDerm - Part 2: Treatment monitoring and specific clinical or comorbid situations.

German S3-Guideline on the treatment of Psoriasis vulgaris, adapted from EuroGuiDerm - Part 2: Treatment monitoring and specific clinical or comorbid situations.

The Diagnostic Value of Blood and Urine IP-10 Test in Children Having Active Tuberculosis or Latent Tuberculosis Infection

Abstract Objective This study aimed to investigate interferon-gamma-inducible protein-10 (IP-10) values in serum and urine in pediatric patients in the diagnosis of active tuberculosis (TB) or latent TB infection (LTBI). It also aimed to investigate whether it can be used as a biomarker to distinguish between active TB and LTBI. Methods Our study comprised active TB (25 patients), LTBI (25 patients), and the “infected” group (50 patients) formed by combining the two groups. As the control group, 37 healthy children were included in the study. TB skin test, plasma IP-10, and urine IP-10 measurements were performed in all patients included in the study. An additional QuantiFERON-TB Gold In-Tube (QFT-GIT) test was performed on patients evaluated as active TB or LTBI. Results Plasma IP-10 levels of the patients in the active TB, LTBI, and the “infected” groups were significantly higher than the control group (p = 0.022, p = 0.028, and p = 0.007, respectively). Urine IP-10 was successful in distinguishing the active TB and “infected” groups from the control group (p = 0.007 and p = 0.047, respectively). Also, in the combined use of the tests, when QFT-GIT and urine IP-10 were positive together, active TB and LTBI could be distinguished (p = 0.044). Urine IP-10 levels were found to be significantly higher in those with pulmonary TB than those with extrapulmonary TB (p = 0.012). Conclusion Our findings suggest that IP-10 can be used as a useful biomarker in the diagnosis of active TB in children.

Differentiating between active and latent tuberculosis with chest computed tomography

PurposeThe purpose of this study was to evaluate the capabilities of chest computed tomography (CT) in distinguishing between active and latent tuberculosis in patients positive for interferon-gamma release assay (IGRA) testing, and to compare the performance of CT with that of quantitative IGRA testing in a low incidence setting. Materials and methodsPatients with latent or active tuberculosis define by an IGRA positive test were retrospectively recruited. Sensitivity, specificity and accuracy were determined for CT variables and quantitative IGRA results. Final diagnosis of active tuberculosis was based on clinical data and microbiological culture. Univariable and multivariable analyses were performed using logistic regression model to identify CT variables associated with the diagnosis of active tuberculosis. ResultsA total of 92 patients with positive IGRA results who underwent CT examination were included. There were 54 men and 38 women with a mean age of 53.5±18 (SD) years (range: 40–68 years). Of them, 22 patients (24%) had positive Mycobacterium tuberculosis culture and 70 (76%) had latent tuberculosis. Among CT variables, consolidation had the greatest sensitivity (77%; 95%CI: 60–95%) and “tree-in-bud” the greatest specificity (97%; 95% CI: 93–100%) for the diagnosis of active tuberculosis. At univariable analysis “tree-in-bud”, splenic calcification and non-calcified lung nodules were the significant variables independently associated with active tuberculosis. At multivariable analysis, the adjusted odds ratio of “tree-in-bud” was 42.91 (95% CI: 5.62–327.42). Using an optimal threshold of 51 spots, quantitative IGRA yielded 64% sensitivity (95% CI: 44–84%) and 61% specificity (95% CI: 50–73%) for the diagnosis of active tuberculosis. ConclusionsIn a low incidence setting, chest CT, especially when “tree-in-bud” pattern is present, is superior to quantitative IGRA testing to identify patients with active tuberculosis among those with positive IGRA testing.

The Evaluation and Validation of Blood-Derived Novel Biomarkers for Precise and Rapid Diagnosis of Tuberculosis in Areas With High-TB Burden

Tuberculosis (TB) remains a highly contagious public health threat. Precise and prompt diagnosis and monitoring of treatment responses are urgently needed for clinics. To pursue novel and satisfied host blood-derived biomarkers, we streamlined a bioinformatic pipeline by integrating differentially expressed genes, a gene co-expression network, and short time-series analysis to mine the published transcriptomes derived from whole blood of TB patients in the GEO database, followed by validating the diagnostic performance of biomarkers in both independent datasets and blood samples of Chinese patients using quantitative real-time PCR (qRT-PCR). We found that four genes, namely UBE2L6 (Ubiquitin/ISG15-conjugating enzyme E2 L6), BATF2 (Basic leucine zipper transcriptional factor ATF-like), SERPING1 (Plasma protease C1 inhibitor), and VAMP5 (Vesicle-associated membrane protein 5), had high diagnostic value for active TB. The transcription levels of these four gene combinations can reach up to 88% sensitivity and 78% specificity (average) for the diagnosis of active TB; the highest sensitivity can achieve 100% by parallel of BATF2 and VAMP5, and the highest specificity can reach 89.5% through a combination of SERPIG1, UBE2L6, and VAMP5, which were significantly higher than 75.3% sensitivity and 69.1% specificity by T-SPOT.TB in the same patients. Quite unexpectedly, the gene set can assess the efficacy of anti-TB response and differentiate active TB from Latent TB infection. The data demonstrated these four biomarkers might have great potency and advantage over IGRAs in the diagnosis of TB.

The TBAg/PHA ratio in T-SPOT.TB assay has high prospective value in the diagnosis of active tuberculosis: a multicenter study in China

BackgroundThe positive rate of pathogenic examination about tuberculosis is low. It is still difficult to achieve early diagnosis for some TB patients. The value of Interferon-gamma release assays (IGRA) in the diagnosis of active tuberculosis remains controversial. The purpose of this multicenter prospective study was to verify and validate the role of TBAg/PHA ratio (TB-specific antigen to phytohaemagglutinin) of T-SPOT.TB assay in diagnosing ATB.MethodsWe prospectively enrolled 2390 suspected pulmonary tuberculosis patients with positive T-SPOT assay results from three tertiary hospitals.ResultsA total of 1549 ATB (active tuberculosis) patients (including 1091 confirmed and 458 probable ATB) and 724 non-tuberculosis (non-TB) patients with positive T-SPOT results were included. The results of this study showed that ESAT-6 and CFP-10 in the T-SPOT.TB assay were significantly higher in the ATB group compared with the non-TB group, while PHA was lower in the ATB group. Results of ESAT-6, CFP-10 and PHA show a certain diagnostic performance, but moderate sensitivity and specificity. The TBAg/PHA ratio, a further calculation of ESAT-6, CFP-10 and PHA in T-SPOT.TB assay showed improved performance in the diagnosis of active Tuberculosis. If using the threshold value of 0.2004, the specificity and sensitivity of TBAg/PHA ratio in distinguishing ATB from non-TB were 92.3% and 74.4%, PPV was 95.4, PLR was 9.6.ConclusionBy recalculating the results of T-SPOT.TB Assay, the TBAg/PHA ratio shows high prospect value in the diagnosis of active tuberculosis in high prediction areas.

The Role of High Resolution Computed Tomography in the Diagnosis of Suspected Sputum Smear Negative Active Pulmonary TB

Objective: To evaluate the diagnostic accuracy of high resolution CT (HrCT) for confirmatory evaluation of patients having active pulmonary TB (PTB) taking Acid-fast bacillus (AFB) culture reporting as standard parameter. Methods: In this cross-sectional (validation) analysis we included 200 patients having chest X-rays and clinical evidence of active TB and sputum smear (SS) –ve results. These patients were recruited from March-2019 to December-2019. Data of patients regarding baseline hematology investigations, sputum smear findings, and presenting symptoms was obtained. Sputum samples were obtained again for each patients, and sent for AFB culture reporting. The HrCT was performed in the radiology unit using a 64 slices CT machine. Results: The mean age of included patients was 53.61 ± 11.96 years. There 81 (40.5%) female and 119 (59.5%) male patients. The proportion of true positive (TP) cases was 79 (39.5%), true negative cases was 96 (48.0%), false positive (FP) cases 11 (5.5%) and false negative (FN) was 14 (7.0%). The HrCT was found to have sensitivity of 84.9%, specificity 89.7%, PPV 87.8% and NPV 87.3%. Conclusion: HrCT is a reliable imaging method for accurately diagnosis the active Pul. TB with a good sensitivity and specificity. In our view, HrCT can be used as a firstline imaging evaluation test for diagnosing pul. TB in suspected patients. Keywords: Pulmonary TB, Microbial culture, high resolution CT.Objective: To evaluate the diagnostic accuracy of high resolution CT (HrCT) for confirmatory evaluation of patients having active pulmonary TB (PTB) taking Acid-fast bacillus (AFB) culture reporting as standard parameter. Methods: In this cross-sectional (validation) analysis we included 200 patients having chest X-rays and clinical evidence of active TB and sputum smear (SS) –ve results. These patients were recruited from March-2019 to December-2019. Data of patients regarding baseline hematology investigations, sputum smear findings, and presenting symptoms was obtained. Sputum samples were obtained again for each patients, and sent for AFB culture reporting. The HrCT was performed in the radiology unit using a 64 slices CT machine. Results: The mean age of included patients was 53.61 ± 11.96 years. There 81 (40.5%) female and 119 (59.5%) male patients. The proportion of true positive (TP) cases was 79 (39.5%), true negative cases was 96 (48.0%), false positive (FP) cases 11 (5.5%) and false negative (FN) was 14 (7.0%). The HrCT was found to have sensitivity of 84.9%, specificity 89.7%, PPV 87.8% and NPV 87.3%. Conclusion: HrCT is a reliable imaging method for accurately diagnosis the active Pul. TB with a good sensitivity and specificity. In our view, HrCT can be used as a firstline imaging evaluation test for diagnosing pul. TB in suspected patients. Keywords: Pulmonary TB, Microbial culture, high resolution CT.

Screening of 20 Mycobacterium tuberculosis sRNAs in plasma for detection of active pulmonary tuberculosis

BackgroundMycobacterium tuberculosis (MTB) sRNAs are abundant. However, the level of MTB sRNA in peripheral blood remains elusive. MethodsTwenty MTB sRNAs annotated in the reference genome of H37Rv were detected in the plasma of 170 active pulmonary tuberculosis patients and 124 healthy people by qRT-PCR detection system. The differential expression of sRNAs were analyzed in two groups. The value of sRNAs for diagnosis of active tuberculosis were evaluated by ROC curve analysis. ResultsEight of the 20 sRNAs (MTS2823, MTS0997, MTS1338, ASdes, G2, C8, mcr15 and MTS1082) were found in at least 50% of the samples detected. The relative expression levels of MTS2823, MTS0997, MTS1338 and ASdes in plasma of tuberculosis patients were statistically higher than those in healthy controls. ROC curve analysis showed that the AUC of MTS0997, MTS1338, MTS2823 and ASdes were 0.8935 (95% CI 0.8109–0.9760), 0.8722 (95% CI 0.7862–0.9581), 0.8208 (95% CI 0.7246–0.9170) and 0.5792 (95% CI 0.4240–0.7344), respectively. The AUC value of combination of MTS0997, MTS1338 and MTS2823 was 0.914 (95% CI 0.8281–0.9926). ConclusionsMTB sRNAs MTS2823, MTS0997 and MTS1338 have the potential to be plasma biomarkers for active pulmonary tuberculosis.

Diagnosis Based on Detection of CXCL10 in Urine as Biomarker for The Determining Diagnosis of Active Lung Tuberculosis

Tuberculosis diagnosis is an important component in decreasing TB incidence and prevalence. Because of the difficulty to collect sputum in some cases, urine specimens are used as it is easier to garner. One of the biomarkers in urine that can be used to diagnose pulmonary TB is IP-10, which can be represented by the CXCL10 gene. The study aims to determine the accuracy of diagnosis based on detection of the CXCL10 gene in urine as a biomarker for the patients with suspected pulmonary TB in Dr. Soetomo Hospital in Surabaya from November 2019 until March 2020. Thus, this is an observative laboratory research with a cross-sectional study. CXCL10 gene was examined using PCR for 36 urine samples, and then, the data, together with the medical records of clinical manifestations of pulmonary TB, GeneXpert MTB /RIF, blood count, and thorax radiograph, were processed using IBM SPSS Statistics 26. The results of the GeneXpert MTB/RIF and thorax radiograph criteria show positive results of pulmonary TB, which were 44.4% and 69.4% respectively. CXCL10 gene was not found in all urine of healthy people (negative), while 2.8% (1/36 samples) positive CXCL10 gene was found in a patient with positive GeneXpert, also with negative clinical manifestations and urine culture. In this study, the accuracy of diagnosis based on detection of the CXCL10 gene in urine for diagnosis of active pulmonary TB was 2.8%. Future research is needed to improve the methods, among them are bigger size of urine samples and clearer medical history of patients.

Blood transcriptomic biomarkers for tuberculosis screening: time to redefine our target populations?

Blood transcriptomic biomarkers for tuberculosis screening: time to redefine our target populations?

Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR.

Tuberculosis (TB) remains a major global threat and diagnosis of active TB ((ATB) both extra-pulmonary (EPTB), pulmonary (PTB)) and latent TB (LTBI) infection remains challenging, particularly in high-burden countries which still rely heavily on conventional methods. Although molecular diagnostic methods are available, e.g., Cepheid GeneXpert, they are not universally available in all high TB burden countries. There is intense focus on immune biomarkers for use in TB diagnosis, which could provide alternative low-cost, rapid diagnostic solutions. In our previous gene expression studies, we identified peripheral blood leukocyte (PBL) mRNA biomarkers in a non-human primate TB aerosol-challenge model. Here, we describe a study to further validate select mRNA biomarkers from this prior study in new cohorts of patients and controls, as a prerequisite for further development. Whole blood mRNA was purified from ATB patients recruited in the UK and India, LTBI and two groups of controls from the UK (i) a low TB incidence region (CNTRLA) and (ii) individuals variably-domiciled in the UK and Asia ((CNTRLB), the latter TB high incidence regions). Seventy-two mRNA biomarker gene targets were analyzed by qPCR using the Roche Lightcycler 480 qPCR platform and data analyzed using GeneSpring™ 14.9 bioinformatics software. Differential expression of fifty-three biomarkers was confirmed between MTB infected, LTBI groups and controls, seventeen of which were significant using analysis of variance (ANOVA): CALCOCO2, CD52, GBP1, GBP2, GBP5, HLA-B, IFIT3, IFITM3, IRF1, LOC400759 (GBP1P1), NCF1C, PF4V1, SAMD9L, S100A11, TAF10, TAPBP, and TRIM25. These were analyzed using receiver operating characteristic (ROC) curve analysis. Single biomarkers and biomarker combinations were further assessed using simple arithmetic algorithms. Minimal combination biomarker panels were delineated for primary diagnosis of ATB (both PTB and EPTB), LTBI and identifying LTBI individuals at high risk of progression which showed good performance characteristics. These were assessed for suitability for progression against the standards for new TB diagnostic tests delineated in the published World Health Organization (WHO) technology product profiles (TPPs).

HIV-Infected Patients Developing Tuberculosis Disease Show Early Changes in the Immune Response to Novel Mycobacterium tuberculosis Antigens.

Background: In individuals living with HIV infection the development of tuberculosis (TB) is associated with rapid progression from asymptomatic TB infection to active TB disease. Sputum-based diagnostic tests for TB have low sensitivity in minimal and subclinical TB precluding early diagnosis. The immune response to novel Mycobacterium tuberculosis in-vivo expressed and latency associated antigens may help to measure the early stages of infection and disease progression and thereby improve early diagnosis of active TB disease.Methods: Serial prospectively sampled cryopreserved lymphocytes from patients of the Swiss HIV Cohort Study developing TB disease (“cases”) and matched patients with no TB disease (“controls”) were stimulated with 10 novel Mycobacterium tuberculosis antigens. Cytokine concentrations were measured in cases and controls at four time points prior to diagnosis of TB: T1-T4 with T4 being the closest time point to diagnosis.Results: 50 samples from nine cases and nine controls were included. Median CD4 cell count at T4 was 289/ul for the TB-group and 456/ul for the control group. Viral loads were suppressed in both groups. At T4 Rv2431c-induced and Rv3614/15c-induced interferon gamma-induced protein (IP)-10 responses and Rv2031c-induced and Rv2346/Rv2347c-induced tumor necrosis factor (TNF)-α responses were significantly higher in cases compared to controls (p < 0.004). At T3 - being up to 2 years prior to TB diagnosis - Rv2031c-induced TNF-α was significantly higher in cases compared to controls (p < 0.004). Area under the receiver operating characteristics (AUROC) curves resulted in an AUC > 0.92 for all four antigen-cytokine pairs.Conclusion: The in vitro Mycobacterium tuberculosis-specific immune response in HIV-infected individuals that progress toward developing TB disease is different from those in HIV-infected individuals that do not progress to developing TB. These differences precede the clinical diagnosis of active TB up to 2 years, paving the way for the development of immune based diagnostics to predict TB disease at an early stage.