7 results on '"Ruhwald, M."'
Search Results
2. Diagnostic benefits of adding EspC, EspF and Rv2348-B to the QuantiFERON Gold In-tube antigen combination.
- Author
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Villar-Hernández R, Blauenfeldt T, García-García E, Muriel-Moreno B, De Souza-Galvão ML, Millet JP, Sabriá F, Sánchez-Montalvá A, Ruiz-Manzano J, Pilarte J, Jiménez MA, Centeno C, Martos C, Molina-Pinargote I, González-Díaz YD, Santiago J, Cantos A, Casas I, Guerola RM, Prat C, Andersen P, Latorre I, Ruhwald M, and Domínguez J
- Subjects
- Adult, Case-Control Studies, Early Diagnosis, Female, Humans, Interferon-gamma Release Tests, Male, Middle Aged, Sensitivity and Specificity, Spain, Tuberculin Test, Tuberculosis immunology, Antigens, Bacterial immunology, Latent Tuberculosis diagnosis, Mycobacterium tuberculosis immunology, Tuberculosis diagnosis
- Abstract
Interferon (IFN)-γ release assays (IGRAs) are used to diagnose latent tuberculosis (TB) infection (LTBI). To improve the accuracy of these tests, different approaches, such as alternative cytokine detection and using different antigens, are considered. Following this purpose, this study aims to evaluate the addition of EspC, EspF and Rv2348-B to those present in the QuantiFERON-TB Gold In-Tube (QFN-G-IT). We included 115 subjects: 74 active TB patients, 17 LTBI individuals and 24 healthy controls. Whole blood samples were collected in QFN-G-IT and in-house tubes containing different combinations of EspC, EspF and Rv2348-B, together with ESAT-6, CFP-10, and TB7.7. After overnight incubation at 37 ºC, plasma was harvested and IFN-γ quantified. IFN-γ levels in the QFN-G-IT and in-house tubes correlated very good (Spearman Rho(r) > 0.86). In-house antigen combinations distinguished healthy individuals from those with active TB and LTBI (specificities and sensitivities higher than 87.5% and 96.3%, respectively [AUC > 0.938]). Adding EspC, EspF and Rv2348-B, increased the sensitivity of the test, being the addition of EspC and Rv2348-B the combination that yielded a higher sensitivity with no specificity loss. Addition of these antigens could improve diagnosis in patients with impaired or immature immune response who are at high risk of developing TB.
- Published
- 2020
- Full Text
- View/download PDF
3. Use of IP-10 detection in dried plasma spots for latent tuberculosis infection diagnosis in contacts via mail.
- Author
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Villar-Hernández R, Latorre I, De Souza-Galvão ML, Jiménez MA, Ruiz-Manzano J, Pilarte J, García-García E, Muriel-Moreno B, Cantos A, Altet N, Millet JP, González-Díaz Y, Molina-Pinargote I, Prat C, Ruhwald M, and Domínguez J
- Subjects
- Adult, Chemokine CXCL10 blood, Contact Tracing, Cytokines blood, Enzyme-Linked Immunosorbent Assay, Female, Humans, Interferon-gamma Release Tests methods, Latent Tuberculosis blood, Male, Postal Service, ROC Curve, Sensitivity and Specificity, Dried Blood Spot Testing methods, Latent Tuberculosis diagnosis
- Abstract
The aim of this study was to test the use of IP-10 detection in dried plasma from contact studies individuals (contacts of smear positive patients), by comparing it with IP-10 and IFN-γ detection in direct plasma, to establish IP-10 detection in DPS as a useful assay for LTBI diagnosis. Whole blood samples were collected from 80 subjects: 12 with active tuberculosis (TB), and 68 from contact studies. The amount of IFN-γ produced by sensitized T cells was determined in direct plasma by QuantiFERON Gold In-Tube test. IP-10 levels were determined in direct and dried plasma by an in-house ELISA. For dried plasma IP-10 determination, two 25 µl plasma drops were dried in Whatman903 filter paper and sent by mail to the laboratory. Regarding TB patients, 100.0%, 91.7% and 75.0% were positive for IFN-γ detection and IP-10 detection in direct and dried plasma, respectively. In contacts, 69.1%, 60.3% and 48.5% had positive results after IFN-γ and IP-10 in direct and dried plasma, respectively. The agreement among in vitro tests was substantial and IP-10 levels in direct and dried plasma were strongly correlated (r = 0.897). In conclusion, IP-10 detection in dried plasma is a simple and safe method that would help improve LTBI management.
- Published
- 2019
- Full Text
- View/download PDF
4. IP-10 dried blood spots assay monitoring treatment efficacy in extrapulmonary tuberculosis in a low-resource setting.
- Author
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Hoel IM, Jørstad MD, Marijani M, Ruhwald M, Mustafa T, and Dyrhol-Riise AM
- Subjects
- Adolescent, Adult, Aged, Antitubercular Agents therapeutic use, Biomarkers blood, Coinfection blood, Enzyme-Linked Immunosorbent Assay, Female, HIV Infections blood, HIV Infections complications, Humans, Longitudinal Studies, Male, Middle Aged, Plasma chemistry, Treatment Outcome, Tuberculosis complications, Tuberculosis drug therapy, Tuberculosis economics, Young Adult, Chemokine CXCL10 blood, Dried Blood Spot Testing economics, Dried Blood Spot Testing methods, Tuberculosis blood
- Abstract
Treatment efficacy is difficult to evaluate in extrapulmonary tuberculosis (EPTB) patients. Interferon-γ inducible protein (IP-)10 has been suggested as a biomarker for response to treatment. We have investigated if IP-10 from dried plasma spots (DPS) or dried blood spots (DBS) can be used in treatment monitoring of EPTB patients in a low-resource setting of Zanzibar. IP-10 levels in plasma, DPS and DBS samples collected before, during (2 months) and after TB treatment of 36 EPTB patients (6 culture and/or Xpert MTB/RIF positive and 30 clinically diagnosed) and 8 pulmonary tuberculosis (PTB) patients, were quantified by an enzyme-linked immunosorbent assay. There was a high positive correlation between IP-10 measured in plasma and DPS and DBS, respectively. We found a significant decline in IP-10 levels from baseline to end of treatment in plasma, DPS and DBS, both in EPTB and PTB patients. The declines were observed already after 2 months in HIV negative patients. In conclusion, the DPS/DBS IP-10 assay allows for easy and manageable monitoring in low-resource settings and our findings suggest that IP-10 may serve as a biomarker for treatment efficacy in EPTB patients, albeit further studies in cohorts of patients with treatment failure and relapse are needed.
- Published
- 2019
- Full Text
- View/download PDF
5. Optimisation of a murine splenocyte mycobacterial growth inhibition assay using virulent Mycobacterium tuberculosis.
- Author
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Jensen C, Lindebo Holm L, Svensson E, Aagaard C, and Ruhwald M
- Subjects
- Animals, Cytokines metabolism, Female, Mice, Mycobacterium bovis immunology, Mycobacterium tuberculosis pathogenicity, Spleen metabolism, T-Lymphocytes immunology, T-Lymphocytes metabolism, Tuberculosis metabolism, Vaccination, Virulence, Mycobacterium tuberculosis immunology, Spleen immunology, Spleen microbiology, Tuberculosis immunology, Tuberculosis microbiology
- Abstract
In the absence of a validated correlate of protection or robust animal models for human tuberculosis, Mycobacterial growth inhibition assays (MGIAs) aim to assess vaccines ability to inhibit mycobacterial growth in-vitro. We optimised a reproducible murine splenocyte MGIA based on in-vitro infection with virulent Mycobacterium tuberculosis (M.tb) Erdman. We identified splenocyte viability as a problem in state-of-art MGIA protocols, which can be improved by simple changes in culture conditions (viability increase from 21% to 46% at last day of culture). The growth inhibitory potential in mice immunised with either BCG, H56:CAF01 or H56:CAF01 administered side-by-side with BCG was significantly better compared to placebo in all groups (0.3 log
10 CFU [±0.2, p = 0.049], 0.5 [±0.2, p = 0.016] and 0.6 [±0.1, p = 0.0007], respectively) corresponding to the levels of in-vivo protection. Unexpectedly the CAF01 adjuvant control group also induced significant growth inhibition of 0.3 log10 CFU (±0.2, p = 0.047). Finally, we explored vaccine-associated T cell effector functions. Despite presence of high levels of vaccine-specific T cells, we found no increase in CD4+ T cell number or cytokine expression profile, nor a difference in cytokine levels in the supernatant after four days culture with or without M.tb. Spontaneous IFN-γ release correlated with growth inhibition levels (p = 0.02), however the cellular source was not found.- Published
- 2017
- Full Text
- View/download PDF
6. Introducing the ESAT-6 free IGRA, a companion diagnostic for TB vaccines based on ESAT-6.
- Author
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Ruhwald M, de Thurah L, Kuchaka D, Zaher MR, Salman AM, Abdel-Ghaffar AR, Shoukry FA, Michelsen SW, Soborg B, Blauenfeldt T, Mpagama S, Hoff ST, Agger EM, Rosenkrands I, Aagard C, Kibiki G, El-Sheikh N, and Andersen P
- Subjects
- Adult, Algorithms, Case-Control Studies, Cohort Studies, Computer Simulation, Female, Humans, Male, Middle Aged, Peptides immunology, ROC Curve, Reproducibility of Results, Tuberculosis immunology, Antigens, Bacterial immunology, Bacterial Proteins immunology, Interferon-gamma Release Tests methods, Tuberculosis Vaccines immunology
- Abstract
There is a need for an improved vaccine for tuberculosis. ESAT-6 is a cardinal vaccine antigen with unique properties and is included in several vaccine candidates in development. ESAT-6 is also the core antigen in the IFN-γ release assays (IGRA) used to diagnose latent infection, rendering IGRA tests unspecific after vaccination. This challenge has prompted the development of a companion diagnostic for ESAT-6 based vaccines, an ESAT-6 free IGRA. We screened a panel of seven potential new diagnostic antigens not recognized in BCG vaccinated individuals. Three highly recognized antigens EspC, EspF and Rv2348c were identified and combined with CFP10 in an ESAT-6 free antigen cocktail. The cocktail was prepared in a field-friendly format, lyophilized with heparin in ready-to-use vacutainer tubes. The diagnostic performance of the ESAT-6 free IGRA was determined in a cross-validation study. Compared IGRA, the ESAT-6 free IGRA induced a comparable magnitude of IFN-γ release, and the diagnostic performance was on par with Quantiferon (sensitivity 84% vs 79%; specificity 99% vs 97%). The comparable performance of the ESAT-6 free IGRA to IGRA suggests potential as companion diagnostic for ESAT-6 containing vaccines and as adjunct test for latent infection.
- Published
- 2017
- Full Text
- View/download PDF
7. IP-10 measured by Dry Plasma Spots as biomarker for therapy responses in Mycobacterium Tuberculosis infection.
- Author
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Tonby K, Ruhwald M, Kvale D, and Dyrhol-Riise AM
- Subjects
- Adult, Aged, Aged, 80 and over, Antitubercular Agents therapeutic use, Biomarkers blood, Cohort Studies, Female, Humans, Male, Middle Aged, Mycobacterium tuberculosis isolation & purification, Mycobacterium tuberculosis pathogenicity, Tuberculosis drug therapy, Tuberculosis microbiology, Chemokine CXCL10 blood, Dried Blood Spot Testing, Enzyme-Linked Immunosorbent Assay, Tuberculosis diagnosis
- Abstract
Tuberculosis (TB) has huge impact on human morbidity and mortality and biomarkers to support rapid TB diagnosis and ensure treatment initiation and cure are needed, especially in regions with high prevalence of multi-drug resistant TB. Soluble interferon gamma inducible protein 10 (IP-10) analyzed from dry plasma spots (DPS) has potential as an immunodiagnostic marker in TB infection. We analyzed IP-10 levels in plasma directly and extracted from DPS in parallel by ELISA from 34 clinically well characterized patients with TB disease before and throughout 24 weeks of effective anti-TB chemotherapy. We detected a significant decline of IP-10 levels in both plasma and DPS already after two weeks of therapy with good correlation between the tests. This was observed both in pulmonary and extrapulmonary TB. In conclusion, plasma IP-10 may serve as an early biomarker for anti-TB chemotherapy responses and the IP-10 DPS method has potential to be developed into a point-of care test for use in resource-limited settings. Further studies must be performed to validate the use of IP-10 DPS in TB high endemic countries.
- Published
- 2015
- Full Text
- View/download PDF
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