scholarly journals Use of Rv0222-Rv2657c-Rv1509 Fusion Protein to Improve the Accuracy of an Antibody ELISA for Extra-Pulmonary Tuberculosis in Humans

Pathogens ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 828
Author(s):  
Yingyu Chen ◽  
Pan Ge ◽  
Kailun Zhang ◽  
Jie Xiang ◽  
Li Zhang ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Fusion Proteins ◽  
Lower Sensitivity ◽  
Epidemic Disease ◽  
Serological Tests ◽  
Extra Pulmonary Tuberculosis ◽  
Pulmonary Tb ◽  
Antibody Elisa ◽  
Extra Pulmonary ◽  
Low Sensitivity

(1) Background: Tuberculosis (TB) in humans is a serious chronic epidemic disease caused by Mycobacterium tuberculosis (M. tb). The diagnosis of TB, especially extra-pulmonary TB (EPTB), is difficult. Isolation of M. tb from culture has a low sensitivity in patients with TB and an even lower sensitivity in cases of EPTB. Although Xpert MTB/RIF assays and serological tests are more sensitive than the above tests, they still lack sensitivity for EPTB diagnosis. (2) Methods: To improve the accuracy of TB diagnosis, a Rv0222-Rv2657c-Rv1509 fusion protein based iELISA was constructed, the diagnosis of TB, pulmonary TB (PTB) and EPTB was then evaluated. Sera of 40 TB patients including 14 with PTB, 14 with EPTB and 12 with no information about the form of TB, and five pneumonia patients were investigated. (3) Results: The sensitivity of the ELISA in TB, PTB and EPTB patients was 80% (95% CI: 64.4, 90.9%), 85.7% (95% CI: 57.2, 98.2%) and 92.8% (95% CI: 66.1, 99.8%), respectively, with a specificity of 70% (95% CI: 53.5, 83.4%). Both the sensitivity and specificity with this fusion protein were higher than for CFP10/ESAT6 (used as reference antigen) fusion protein (71.4%; 95% CI: 41.9, 91.6%, and 67.5%; 95% CI: 50.9, 81.4%), respectively, in cases of EPTB. All pneumonia patients’ sera tested negative in both ELISAs. (4) Conclusion: use of these new fusion proteins as antigens in serological assays has the potential to improve the diagnosis of all forms of TB in humans, especially EPTB.

Usefulness of GeneXpert MTB/RIF in the diagnosis of extra-pulmonary tuberculosis

2021 ◽  
Vol 11 (2) ◽  
pp. 121-124
Author(s):  
Nasreen Islam ◽  
Mohammad Delwar Hossain ◽  
Muhammad Abdur Rahim ◽  
Jamal Uddin Ahmed ◽  
Muhammad Kamrul Amin ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Pleural Fluid ◽  
Past History ◽  
Gastric Lavage ◽  
Extra Pulmonary Tuberculosis ◽  
Pulmonary Tb ◽  
Male Predominance ◽  
Bacteriological Diagnosis ◽  
Extra Pulmonary ◽  
Tracheal Aspirates

Background: Diagnosis of extra-pulmonary tuberculosis (TB) is often delayed because of diverse clinical presentations and difficulties in establishing the bacteriological diagnosis. This study aimed to evaluate usefulness of GeneXpert MTB/RIF in the diagnosis of extra-pulmonary TB in Bangladeshi patients. Methods: This cross-sectional study was done in BIRDEM General Hospital, Dhaka, Bangladesh from 2013 to 2016 as a part of Bangladesh Diabetic Somiti (BADAS)-USAID-TB Care-II project. Representative samples from 590 clinically suspected extra-pulmonary TB cases were tested for GeneXpert MTB/RIF along with conventional methods. Results: Total patients were 590 [mean age 43.9 (range 1-95) years] with male predominance (326, 55.3%). Most (513, 86.9%) patients were diabetic and new (574, 97.3%) TB suspects; while 16 (2.7%) patients had past history of TB. Common samples were pleural fluid (125, 21.2%), urine (110, 18.6%), cerebrospinal fluid (CSF) (91, 15.4%), pus (82, 13.9%), tracheal aspirates (57, 9.7%), ascitic fluid (45, 7.6%), gastric lavage (31, 5.3%), broncho-alveolar lavage (BAL) (18, 3.1%), lymph node aspirates (11, 1.9%) and synovial fluid (8, 1.4%). Among 590 samples, 68 (11.5%) were positive for Mycobacterium tuberculosis. Diagnostic yield was common for lymph nodes (4/7, 57.1%), pus (25/82, 30.5%), BAL (4/18, 22.2%), tracheal aspirates (8/57, 14.0%), urine (7/110, 6.4%), CSF (6/91, 6.6%) and pleural fluid (7/125, 5.6%). Of the 68 GeneXpert MTB/RIF positive samples, 52 (76.1%) were rifampicin sensitive, 16 (23.9%) showed intermediate sensitivity and none of the samples was resistant to rifampicin. Conclusions: GeneXpert MTB/RIF appeared as useful tool for diagnosing extra-pulmonary TB. Birdem Med J 2021; 11(2): 121-124

scholarly journals Extra pulmonary tuberculosis manifestation in head and neck and its diagnosis with recent tools like cartridge based nucleic acid amplification test

2021 ◽  
Vol 7 (5) ◽  
pp. 829
Author(s):  
Richa Vatsyayan ◽  
Kavita Sachdeva
Keyword(s):  
Nucleic Acid ◽  
Pulmonary Tuberculosis ◽  
Head And Neck ◽  
Clinical Manifestations ◽  
Nucleic Acid Amplification Test ◽  
Nucleic Acid Amplification ◽  
Extra Pulmonary Tuberculosis ◽  
Pulmonary Tb ◽  
Extra Pulmonary ◽  
Sensitivity Specificity

<p class="abstract"><strong>Background:</strong> The study aimed to assess the clinical manifestations of extra-pulmonary tuberculosis (head and neck) in diagnosed pulmonary tuberculosis (TB) patients and without pulmonary TB and to determine the sensitivity and specificity of cartridge based nucleic acid amplification test (CBNAAT) in the diagnosis of extra-pulmonary TB.</p><p class="abstract"><strong>Methods:</strong> The study was conducted as an observational study at the Department of Otorhinolaryngology, tertiary care center, Jabalpur (Madhya Pradesh) for a period of 18 months i.e. from 01 March 2019 to 31 August 2020 on 30 patients presenting with lesions in ear, nose and throat (ENT), head and neck region. All necessary investigations with TB workup were done. Sputum samples were analyzed by CBNAAT on Xpert- <em>Mycobacterium tuberculosis</em> complex/resistance to rifampin (MTB/RIF). Patients were treated accordingly. Appropriate surgical intervention was done in necessary cases. Using statistical analysis, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy was calculated for CBNAAT.  </p><p class="abstract"><strong>Results:</strong> Neck swelling and ear discharge were the most common features observed in 66.7% of cases. In cases with pulmonary TB, systemic features were the most predominant features whereas in cases with extra-pulmonary TB, local features were predominant. Sensitivity, specificity, PPV and NPV of CBNAAT were documented to be 85%, 50%, 89.5% and 40% respectively.</p><p class="abstract"><strong>Conclusions:</strong> TB of the head and neck are commonly encountered in clinical practice. The patients usually present with variable clinical manifestations depending upon the site of involvement. The middle ear is the most common site affected by head and neck tuberculosis. CBNAAT is a sensitive tool for the diagnosis of extra-pulmonary TB with and without associated pulmonary TB.</p>

scholarly journals Pulmonary involvement in patients presenting with extra-pulmonary tuberculosis: thinking beyond a normal chest x-ray

2014 ◽  
Vol 6 (1) ◽  
pp. 64 ◽  
Author(s):  
Samantha Herath ◽  
Christopher Lewis
Keyword(s):  
Public Health ◽  
Pulmonary Tuberculosis ◽  
Pulmonary Involvement ◽  
Extra Pulmonary Tuberculosis ◽  
Pulmonary Tb ◽  
X Ray ◽  
Normal Chest ◽  
Chest X Ray ◽  
Extra Pulmonary ◽  
Hiv Negative

INTRODUCTION: Recognition of pulmonary involvement in extra-pulmonary tuberculosis (TB) may be an important public health issue, as smear-negative pulmonary TB is responsible for about 17% of new infections. Pulmonary TB can be present despite a normal chest x-ray (CXR), even in human immunodeficiency virus (HIV)–negative patients. In this retrospective clinical audit, we reviewed a case series of HIV-negative patients with extra-pulmonary tuberculosis to identify the proportion with concurrent pulmonary TB despite an unremarkable CXR. METHODS: Clinical notes, microbiology results and CXR reports were reviewed from consecutive patients treated at Auckland City Hospital for extra-pulmonary TB from January 2007 to July 2010. RESULTS: Of the sample of 103 patients with extra-pulmonary TB, the majority of patients were born in an Asian country (n=70; 68%). The commonest presentation of extra-pulmonary TB was lymphadenopathy (n=51; 50%), followed by pleural (n=24; 23%) and bone (n=6; 6%) disease. Extra-pulmonary TB was diagnosed by biopsy or excision of the extra-pulmonary site in the majority (n=74; 72%), and by sputum testing alone in 26 (25%). The majority had CXR abnormalities (n=76; 74%). In the group with a normal CXR (n=27), 55% (n=15) had sputum cultures performed. In total, 18% (n=5) of patients with extra-pulmonary TB and a normal CXR had pulmonary TB, of whom two were smear positive. DISCUSSION: In patients with extra-pulmonary TB, sputum testing should be considered to detect concurrent pulmonary TB even if a CXR is normal, especially in immunosuppressed or symptomatic patients. This may aid diagnosis and determine infectivity and consequent public health action. KEYWORDS: Chest x-ray; sputum; tuberculosis; tuberculosis, pulmonary

scholarly journals A study of extra-pulmonary tuberculosis and its outcome

2017 ◽  
Vol 4 (1) ◽  
pp. 209 ◽  
Author(s):  
Ravikumar P. ◽  
Priyadarshini Bai G.
Keyword(s):  
Alcohol Abuse ◽  
Pulmonary Tuberculosis ◽  
Common Site ◽  
Extra Pulmonary Tuberculosis ◽  
Tuberculosis Patients ◽  
Pulmonary Tb ◽  
Medical College ◽  
Extra Pulmonary ◽  
One Year ◽  
Tuberculosis Disease

Background: Pulmonary tuberculosis being the predominant manifestation of the disease extra-pulmonary sites can also involve as a result of dissemination from a chief focus. Extra- pulmonary tuberculosis is more common in HIV cases. The present study aims to determine the presentation and outcome of patients with extra-pulmonary tuberculosis treated with DOTS and to assess any difference in outcome of treatment in HIV positive extra-pulmonary tuberculosis.Methods: Data was collected from cases of tuberculosis patients diagnosed and treated under DOTS at Sri Siddhartha Medical College for the period of one year (during 2015). We evaluated extra-pulmonary cases and recorded sites of involvement in order of frequency. We also studied treatment outcome by recording as per definitions given by the WHO and also evaluated any difference in outcome of extra-pulmonary tuberculosis disease with HIV co-infection.Results: Extra-pulmonary cases accounted for 30.5% of total TB cases. Among 224 cases of extra-pulmonary TB studied, 136 (60.7%) were males and 88 (39.3%) were females. Most common site of extra-pulmonary tuberculosis was pleura (29.9%) followed by meninges (22.5%), abdomen (19.6%) and lymph node (10.7%) tuberculosis. Among these patients 82.2% completed treatment, 7.5% were defaulted, 9.9% died and 0.4% treatment failure. The most common reason for default was irregular treatment (29.5%) followed by alcohol abuse (23.5%). Among 8 HIV reactive patients, 5 patients completed treatment and the remaining 3 patients died during the course of treatment.Conclusions: Extra-pulmonary Tuberculosis accounts for 30.5% of the total cases studied. Pleura is most common site of extra-pulmonary TB in our study. Treatment irregularities and alcohol abuse are the two most common reasons for default.   Co-infection with HIV seems to have a poor outcome on patients with extra-pulmonary TB and needs to be studied in large number of samples.

CORRELATION OF CYTOMORPHOLOGY WITH ZN AND AR POSITIVITY ON FNAC IN THE DIAGNOSIS OF EXTRA PULMONARY TUBERCULOSIS

2021 ◽  
pp. 7-8
Author(s):  
Sana Umar ◽  
Syed Shamshad Ahmad ◽  
Kafil Akhtar ◽  
Zuber Ahmad
Keyword(s):  
Pulmonary Tuberculosis ◽  
Diagnostic Yield ◽  
Extrapulmonary Tuberculosis ◽  
Invasive Procedure ◽  
Epithelioid Cell ◽  
Chi Square ◽  
Extra Pulmonary Tuberculosis ◽  
Epithelioid Cell Granuloma ◽  
Extra Pulmonary ◽  
Low Sensitivity

BACKGROUND: FNAC is a minimally invasive procedure with a signicant diagnostic role in extrapulmonary tuberculosis. ZN Stain for demonstration of mycobacterium tuberculosis is extensively used. However, it has low sensitivity. Fluorochrome stain like Auramine- Rhodamine (AR) appear to be more likely to detect tubercular bacilli than ZN stain and also reduces the examination time. To study the OBJECTIVES: correlation of cytomorphology of extrapulmonary tuberculosis(EPTB) obtained on FNAC with ZN and AR positivity. MATERIALMETHODS: A total of 250 patients were taken, that were referred to the Department of Pathology, Jawaharlal Nehru Medical College, AMU, Aligarh from October 2015 to November 2017. Samples were collected by FNA and smears prepared were stained with H&E, ZN and AR stain. Smears were observed for positivity of Acid Fast Bacilli (AFB). Culture was taken as gold standard. RESULTS: The most common site of extra pulmonary tuberculosis was cervical lymph node seen in 76.4% cases and the most common cytomorphological pattern on FNAC was epithelioid cell granuloma with necrosis seen in 86.4% cases. The sensitivity of AR stain in picking up AFB was found to be 63.4% and the specicity was 81.9%, whereas ZN stain had a low sensitivity of 45.3% but had a high specicity of 87.9%. Statistically signicant difference between the two stains was seen on applying chi square test (p<0.001). Cohen's kappa coefcient for ZN vs AR stain was 0.65 and the strength of agreement between the two stains was substantial CONCLUSION: AR stain is more sensitive than ZN stain in diagnosing extrapulmonary tuberculosis and on combining it with cytomorphology it can help increase the diagnostic yield.

Validity of a Serological Diagnostic Kit for SARS-CoV-2 Available in Iran

2020 ◽  
Vol 23 (9) ◽  
pp. 629-632
Author(s):  
Hamid Reza Shamsollahi ◽  
Mostafa Amini ◽  
Shaban Alizadeh ◽  
Saharnaz Nedjat ◽  
Ali Akbari-Sari ◽  
...  
Keyword(s):  
Diagnostic Techniques ◽  
Effective Control ◽  
Molecular Technique ◽  
Size Estimation ◽  
Medical Sciences ◽  
Serological Tests ◽  
Standard Case ◽  
Epidemic Size ◽  
Low Sensitivity ◽  
Negative Controls

Background: The Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemic broke out in December 2019 and is now characterized as a pandemic. Effective control of this infectious disease requires access to diagnostic techniques, for both case finding and epidemic size estimation. The molecular technique is routinely used worldwide. Although it is the "standard" case detection and management method, it has its own shortcomings. Thus, some easy-to-use rapid serological tests have been developed. Methods: One hundred and fourteen positive RT-PCR-diagnosed patients were tested by VivaDiag Kit, a brand of rapid serological kits available in hospitals affiliated to Tehran University of Medical Sciences (TUMS), Tehran, Iran. Frozen serum specimens taken from healthy people in summer and fall 2019 were also tested as negative controls. Results: Test sensitivity was 47.9% (95% confidence interval [CI]: 38.8-56.9) for IgM and 47.0% (95% CI: 38.0–56.0) for IgG. There was no difference between IgG and IgM seropositivity except in one case. Specificity was calculated as 99.0% (95% CI: 96.4–99.9) for IgM and of 100.0% (95% CI: 0.98.2–100.0) for IgG. Sensitivity was higher in men and older participants. Conclusion: This test can be used for epidemiological investigations, especially for the estimation of the level of infection in the community, after it is properly corrected for sensitivity and specificity. The low sensitivity could be attributed to the technical limitations of the kit or low levels of antibodies after infection. The different sensitivity in age and sex groups supports the hypothesis that different people show different immune responses to this virus.

scholarly journals RGD4C peptide mediates anti-p21Ras scFv entry into tumor cells and produces an inhibitory effect on the human colon cancer cell line SW480

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Chen-Chen Huang ◽  
Fang-Rui Liu ◽  
Qiang Feng ◽  
Xin-Yan Pan ◽  
Shu-Ling Song ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Cell Membrane ◽  
Fusion Protein ◽  
Tumor Cell ◽  
Tumor Cells ◽  
Fusion Proteins ◽  
Inhibitory Effect ◽  
Endosomal Escape ◽  
Antitumor Effects ◽  
Sw480 Cells

Abstract Background We prepared an anti-p21Ras scFv which could specifically bind with mutant and wild-type p21Ras. However, it cannot penetrate the cell membrane, which prevents it from binding to p21Ras in the cytoplasm. Here, the RGD4C peptide was used to mediate the scFv penetration into tumor cells and produce antitumor effects. Methods RGD4C-EGFP and RGD4C-p21Ras-scFv recombinant expression plasmids were constructed to express fusion proteins in E. coli, then the fusion proteins were purified with HisPur Ni-NTA. RGD4C-EGFP was used as reporter to test the factors affecting RGD4C penetration into tumor cell. The immunoreactivity of RGD4C-p21Ras-scFv toward p21Ras was identified by ELISA and western blotting. The ability of RGD4C-p21Ras-scFv to penetrate SW480 cells and colocalization with Ras protein was detected by immunocytochemistry and immunofluorescence. The antitumor activity of the RGD4C-p21Ras-scFv was assessed with the MTT, TUNEL, colony formation and cell migration assays. Chloroquine (CQ) was used an endosomal escape enhancing agent to enhance endosomal escape of RGD4C-scFv. Results RGD4C-p21Ras-scFv fusion protein were successfully expressed and purified. We found that the RGD4C fusion protein could penetrate into tumor cells, but the tumor cell entry of was time and concentration dependent. Endocytosis inhibitors and a low temperature inhibited RGD4C fusion protein endocytosis into cells. The change of the cell membrane potential did not affect penetrability. RGD4C-p21Ras-scFv could penetrate SW480 cells, effectively inhibit the growth, proliferation and migration of SW480 cells and promote this cells apoptosis. In addition, chloroquine (CQ) could increase endosomal escape and improve antitumor activity of RGD4C-scFv in SW480 cells. Conclusion The RGD4C peptide can mediate anti-p21Ras scFv entry into SW480 cells and produce an inhibitory effect, which indicates that RGD4C-p21Ras-scFv may be a potential therapeutic antibody for the treatment of ras-driven cancers.

scholarly journals Seasonality in extra-pulmonary tuberculosis notifications in Germany 2004-2014- a time series analysis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Tanja Charles ◽  
Matthias Eckardt ◽  
Basel Karo ◽  
Walter Haas ◽  
Stefan Kröger
Keyword(s):  
Time Series ◽  
Time Series Analysis ◽  
Disease Onset ◽  
Age Groups ◽  
Age Group ◽  
Pulmonary Tb ◽  
Series Analysis ◽  
Disease Site ◽  
Extra Pulmonary

Abstract Background Seasonality in tuberculosis (TB) has been found in different parts of the world, showing a peak in spring/summer and a trough in autumn/winter. The evidence is less clear which factors drive seasonality. It was our aim to identify and evaluate seasonality in the notifications of TB in Germany, additionally investigating the possible variance of seasonality by disease site, sex and age group. Methods We conducted an integer-valued time series analysis using national surveillance data. We analysed the reported monthly numbers of started treatments between 2004 and 2014 for all notified TB cases and stratified by disease site, sex and age group. Results We detected seasonality in the extra-pulmonary TB cases (N = 11,219), with peaks in late spring/summer and troughs in fall/winter. For all TB notifications together (N = 51,090) and for pulmonary TB only (N = 39,714) we did not find a distinct seasonality. Additional stratified analyses did not reveal any clear differences between age groups, the sexes, or between active and passive case finding. Conclusion We found seasonality in extra-pulmonary TB only, indicating that seasonality of disease onset might be specific to the disease site. This could point towards differences in disease progression between the different clinical disease manifestations. Sex appears not to be an important driver of seasonality, whereas the role of age remains unclear as this could not be sufficiently investigated.

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