Comparative evaluation of innovative diagnostic tests for latent and active TВ infection in children

2014 ◽  
Vol 5 (3) ◽  
pp. 46-50 ◽  
Author(s):  
Marina Eduardovna Lozovskaya ◽  
Vyacheslav Valeryevich Belushkov ◽  
Olga Petrovna Gurina ◽  
Yelena Borisovna Vasilyeva ◽  
Lyudmila Vladimirovna Klochkova
Keyword(s):  
Test System ◽  
Mantoux Test ◽  
In Vitro Tests ◽  
Lower Sensitivity ◽  
Additional Information ◽  
Wide Clinical Application ◽  
Latent Tb ◽  
Latent Tb Infection ◽  
Quantiferon Test

The results of three new tests based on antigens CFP-10 аnd ESAT-6 were studied in 50 children: intradermal Diaskintest and tests in vitro QUANTIFERON and Tubinferon. Diaskintest and Tubinferon are developed and produced in the Russian Federation. QUANTIFERON test is performed by Cellestis (Australia). 20 children had active TB, 22 children had latent TB infection, and 8 were not infected by M. tuberculosis. It has been established that while the frequency of coincidence of the results is high (66 %), in some situations, tests may react differently and give additional information when used together in difficult diagnostic cases. In vitro tests were more sensitive compared with Diaskinintest in immunopathological conditions. Tubinferon test had higher sensitivity in latent TB infection (40.9 %) compared to Diaskintest (22.7 %) and QUANTIFERON test (31,8 %), but lower sensitivity in developed tuberculosis - 60, 80 and 85 % respectively. All three tests are more informative in TB disease than in latent TB infection. An important advantage of the Tubinferon test is the ability to evaluate in vitro postvaccinal allergy, due to presence of the sample with the tuberculin. This can be used in the differential diagnostic between tuberculosis and generalized BCG infection, including children with HIV. Mantoux test may be more effective then Diaskintest in children who was not vaccinated with BCG. Tubinferon test system deserves wide clinical application, further study and development.

scholarly journals Tuberculosis infection in children with negative reactions to the diaskintest

2019 ◽  
Vol 10 (3) ◽  
pp. 37-44
Author(s):  
Yulia A. Yarovaya ◽  
Marina E. Lozovskaya ◽  
Ludmila V. Klochkova ◽  
Elena B. Vasilieva ◽  
Gennadii A. Stepanov ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
Tuberculosis Infection ◽  
Mantoux Test ◽  
In Vitro Tests ◽  
Number Of Patients ◽  
Duration Of Infection ◽  
Group 2 ◽  
Tuberculosis Disease ◽  
Quantiferon Test

Variants of the course of tuberculosis infection in 54 children from two to 14 years old, negatively reacting to a sample with an allergen tuberculosis recombinant (Diaskintest) were analyzed. There were 3 groups: 1st – 27 children infected with Mycobacterium tuberculosis (MBT), 50. 0% of cases; 2nd – 16 children with newly diagnosed residual post-tuberculosis changes (OPTI), 29. 6% of cases; 3rd group – 11 patients with active tuberculosis, 20. 4% of cases. Methods of examination: intradermal Mantoux test with 2TE and Diaskintest, according to the testimony of a number of patients in vitro tests: QuantiFERON test (QFT), – SPOT test.TV, multispiral computed tomography, bacteriological, molecular genetic methods of investigation on MBT. The method of mass tuberculin diagnostics revealed 70. 4 ± 8. 8% of children of the 1st group, 93. 8 ± 4. 7% of the 2nd group and 54. 6 ± 15. 0% of children of the 3rd group. The duration of infection with MBT in children was different and was less than 1 year in children of the 1st and 2nd groups – 51. 9 ± 9. 6% and 43. 8 ± 12. 4% of cases, respectively, which was significantly more frequent than in patients of the 3rd group (18. 2 ± 11. 6% of cases). Tuberculosis disease occurred in the form of complicated forms of the primary period-in 45. 5 ± 15. 0 % of cases, uncomplicated forms – in 27. 3 ± 3. 4% of cases, generalized lesions – in 27. 3 ± 13. 4% of cases. Diagnosis of a specific lesion occurred equally in the manifest phases of inflammation: infiltration, infiltration and decay (45. 5 ± 15. 0% of cases), and in the phase of ongoing reverse development (incomplete calcination – in 45. 5 ± 15. 0% of cases), one child had a combination of infiltration and calcination phases (9. 1 ± 8. 7% of cases). Residual posttuberculosis changes in children of group 2 were more often formed in the form of calcifications in the organs of the thoracic cavity – in 87. 5% of cases, in 12. 5 ± 8. 3% of patients OPTI was formed by the formation of seals. Conclusion: in children with negative reactions to the Diaskintest requires individual comprehensive diagnosis of tuberculosis infection.

scholarly journals Recombinant in vitro test T-SPOT.TB as a screening method for early diagnosis of tuberculosis infection

2020 ◽  
Vol 98 (4) ◽  
pp. 48-52
Author(s):  
E. P. Eremenko ◽  
E. A. Borodulina ◽  
I. A. Sergeeva ◽  
D. A. Kudlay ◽  
B. E. Borodulin
Keyword(s):  
Screening Method ◽  
Latent Tuberculosis ◽  
Tuberculosis Infection ◽  
Mantoux Test ◽  
Skin Tests ◽  
In Vitro Tests ◽  
In Vitro Test ◽  
Healthy Children ◽  
Vitro Test

In addition to standard skin tests (Mantoux test with 2 TU PPD-L and diaskintest) for the diagnosis of tuberculosis infection, in vitro tests are used. One of these tests is T-SPOT.TB being more widely used in recent years.The objective: to evaluate the effectiveness of T-SPOT.TB test for early detection of tuberculosis infection in children and adolescents in Samara Region.Subjects and methods. From 2016 to 2019, results of T-SPOT.TB tests performed in 596 children aged 2 to 17 years inclusive were analyzed; those children had no immunodiagnosis of tuberculosis infection using skin tests since their parents refused to have it.Results. It was found out that the major reason for refusing skin tests was the “fear” of visiting a TB dispensary if the result had been positive — 38.43% (n = 229). The latent tuberculosis infection according to the results of T-SPOT.TB among children with concomitant pathology made 2.6%, among healthy children – 0.7%.Conclusion. T-SPOT.TB test may be used as an alternative method for diagnosis of tuberculosis infection, should the parent refuse to have skin tests. In children with concomitant pathology, T-SPOT.TB test can serve as a leading method for immunodiagnosis of tuberculosis.The authors state that they have no conflict of interests.

scholarly journals Multiple-endpoint in vitro carcinogenicity test in human cell line TK6 distinguishes carcinogens from non-carcinogens and highlights mechanisms of action

2020 ◽  
Author(s):  
Katherine E. Chapman ◽  
Eleanor C. Wilde ◽  
Fiona M. Chapman ◽  
Jatin R. Verma ◽  
Ume-Kulsoom Shah ◽  
...  
Keyword(s):  
Cell Line ◽  
Human Cell Line ◽  
Test System ◽  
In Vitro Tests ◽  
Correct Identification ◽  
In Vitro Test ◽  
Human Lymphoblastoid Cell ◽  
Carcinogenic Potential

Abstract Current in vitro genotoxicity tests can produce misleading positive results, indicating an inability to effectively predict a compound’s subsequent carcinogenic potential in vivo. Such oversensitivity can incur unnecessary in vivo tests to further investigate positive in vitro results, supporting the need to improve in vitro tests to better inform risk assessment. It is increasingly acknowledged that more informative in vitro tests using multiple endpoints may support the correct identification of carcinogenic potential. The present study, therefore, employed a holistic, multiple-endpoint approach using low doses of selected carcinogens and non-carcinogens (0.001–770 µM) to assess whether these chemicals caused perturbations in molecular and cellular endpoints relating to the Hallmarks of Cancer. Endpoints included micronucleus induction, alterations in gene expression, cell cycle dynamics, cell morphology and bioenergetics in the human lymphoblastoid cell line TK6. Carcinogens ochratoxin A and oestradiol produced greater Integrated Signature of Carcinogenicity scores for the combined endpoints than the “misleading” in vitro positive compounds, quercetin, 2,4-dichlorophenol and quinacrine dihydrochloride and toxic non-carcinogens, caffeine, cycloheximide and phenformin HCl. This study provides compelling evidence that carcinogens can successfully be distinguished from non-carcinogens using a holistic in vitro test system. Avoidance of misleading in vitro outcomes could lead to the reduction and replacement of animals in carcinogenicity testing.

scholarly journals In vitro Mycobacterial Growth Inhibition in South Korean Adults With Latent TB Infection

2019 ◽  
Vol 10 ◽  
Author(s):  
Hyejon Lee ◽  
Jungho Kim ◽  
Young Ae Kang ◽  
Deok Ryun Kim ◽  
Bora Sim ◽  
...  
Keyword(s):  
Growth Inhibition ◽  
South Korean ◽  
Korean Adults ◽  
Latent Tb ◽  
Latent Tb Infection ◽  
Mycobacterial Growth

scholarly journals Effect of isoniazid on antigen-specific interferon-γ secretion in latent tuberculosis

2014 ◽  
Vol 45 (2) ◽  
pp. 473-482 ◽  
Author(s):  
Martha Torres ◽  
Lourdes García-García ◽  
Pablo Cruz-Hervert ◽  
Heinner Guio ◽  
Claudia Carranza ◽  
...  
Keyword(s):  
Latent Tuberculosis ◽  
Peptide Pool ◽  
Interferon Γ ◽  
Treatment Regimes ◽  
Latent Tb ◽  
Ifn Γ ◽  
New Treatment ◽  
Latent Tb Infection ◽  
Culture Filtrate Protein

Treatment of persons with latent tuberculosis (TB) infection at greatest risk of reactivation is an important component of TB control and elimination strategies. Biomarkers evaluating the effectiveness of treatment of latent TB infection have not yet been identified. This information would enhance control efforts and assist the evaluation of new treatment regimes.We designed a two-group, two-arm, randomised clinical study of tuberculin skin test-positive participants: 26 with documented contact with TB patients and 34 with non-documented contact. Participants in each group were randomly assigned to the immediate- or deferred-isoniazid treatment arms. Assays ofin vitrointerferon (IFN)-γ secretion in response to recombinant Rv1737 and overlapping synthetic peptide pools from various groups of immunodominant proteins were performed.During isoniazid therapy, a significant increase from baseline in the proportion of IFN-γ responders to the 10-kDa culture filtrate protein, Rv2031, Rv0849, Rv1986, Rv2659c, Rv2693c and the recombinant Rv1737 protein was observed (p⩽0.05). The peptide pool of Rv0849 and Rv1737 recombinant proteins induced the highest percentage of IFN-γ responders after isoniazid therapy.Thein vitroIFN-γ responses to these proteins might represent useful markers to evaluate changes associated with treatment of latent TB infection.

scholarly journals Characterization of Host and Microbial Determinants in Individuals with Latent Tuberculosis Infection Using a Human Granuloma Model

mBio ◽  
2015 ◽  
Vol 6 (1) ◽  
Author(s):  
Evelyn Guirado ◽  
Uchenna Mbawuike ◽  
Tracy L. Keiser ◽  
Jesus Arcos ◽  
Abul K. Azad ◽  
...  
Keyword(s):  
Immune Response ◽  
Mycobacterium Tuberculosis ◽  
Immune Status ◽  
Host Immune Response ◽  
Granuloma Formation ◽  
Content Type ◽  
Transcriptional Signature ◽  
Latent Tb ◽  
Latent Tb Infection

ABSTRACTGranulomas sit at the center of tuberculosis (TB) immunopathogenesis. Progress in biomarkers and treatment specific to the human granuloma environment is hindered by the lack of a relevant and tractable infection model that better accounts for the complexity of the host immune response as well as pathogen counterresponses that subvert host immunity in granulomas. Here we developed and characterized anin vitrogranuloma model derived from human peripheral blood mononuclear cells (PBMCs) and autologous serum. Importantly, we interrogated this model for its ability to discriminate between host and bacterial determinants in individuals with and without latent TB infection (LTBI). By the use of this model, we provide the first evidence that granuloma formation, bacterial survival, lymphocyte proliferation, pro- and anti-inflammatory cytokines, and lipid body accumulation are significantly altered in LTBI individuals. Moreover, we show a specific transcriptional signature ofMycobacterium tuberculosisassociated with survival within human granuloma structures depending on the host immune status. Our report provides fundamentally new information on how the human host immune status and bacterial transcriptional signature may dictate early granuloma formation and outcome and provides evidence for the validity of the granuloma model and its potential applications.IMPORTANCEIn 2012, approximately 1.3 million people died from tuberculosis (TB), the highest rate for any single bacterial pathogen. The long-term control of TB requires a better understanding ofMycobacterium tuberculosispathogenesis in appropriate research models. Granulomas represent the characteristic host tissue response to TB, controlling the bacilli while concentrating the immune response to a limited area. However, complete eradication of bacteria does not occur, sinceM. tuberculosishas its own strategies to adapt and persist. Thus, theM. tuberculosis-containing granuloma represents a unique environment for dictating both the host immune response and the bacterial response. Here we developed and characterized anin vitrogranuloma model derived from blood cells of individuals with latent TB infection that more accurately defines the human immune response and metabolic profiles ofM. tuberculosiswithin this uniquely regulated immune environment. This model may also prove beneficial for understanding other granulomatous diseases.

Prospective evaluation of latent tuberculosis with interferon-γ release assays in drug and alcohol abusers

2009 ◽  
Vol 137 (9) ◽  
pp. 1342-1347 ◽  
Author(s):  
I. RIVAS ◽  
I. LATORRE ◽  
A. SANVISENS ◽  
J. DOMÍNGUEZ ◽  
J. TOR ◽  
...  
Keyword(s):  
Latent Tuberculosis ◽  
Interferon Γ ◽  
Alcohol Abusers ◽  
High Prevalence ◽  
History Of ◽  
Latent Tb ◽  
Latent Tb Infection ◽  
Drug And Alcohol ◽  
Good Agreement

SUMMARYIn vitro tests have been developed for the diagnosis of tuberculosis (TB) infection. The objective was to analyse latent TB infection in drug and alcohol abusers through two interferon-γ techniques. One hundred and thirty-nine patients were admitted between February 2006 and May 2007. Mean age was 39·8 years [31% HIV positive]. The enzyme immunoassay (EIA) and enzyme-linked immunospot (ELISPOT) interferon-γ assays were positive in 34% of patients with an agreement of 83% (κ=0·63). Tuberculin skin test (TST) was positive in 29% of patients and the agreement of TST with EIA and ELISPOT interferon-γ assays was 85% (κ=0·62) and 83% (κ=0·57), respectively. Almost 50% of patients with history of TB had a positive in vitro test. In conclusion, we observed a high prevalence of latent TB and good agreement between the new in vitro tests that otherwise may continue to be positive long after developing TB disease.

An Investigation of the Assay of Heparin Based on Its Activity in Vivo

1967 ◽  
Vol 18 (01/02) ◽  
pp. 150-160 ◽  
Author(s):  
W. E Rezansoff ◽  
L. B Jaques
Keyword(s):  
Lipolytic Activity ◽  
Area Under The Curve ◽  
Test System ◽  
Assay Method ◽  
In Vitro Tests ◽  
Clotting Time ◽  
Oil Emulsion ◽  
Effective Doses

SummaryThe potency of different heparin preparations was investigated in vivo, by measuring the Lee and White clotting time, lipolytic activity and partial thromboplastin time of blood samples drawn at intervals after the intravenous injection of heparin into the anesthetized dog. The blood clotting time in log minutes, the thromboplastin time in log seconds, and the plasma lipolytic activity measured as the decrease in optical density of a synthetic cocuont oil emulsion per unit of time after incubation with postheparin plasma, were plotted against time after injection. A linear relation was obtained between the dose of heparin and the response measured as the area under the curve.The in vivo responses for 6 heparin preparations at 3 dose levels were compared with those for a reference heparin. The potency of each relative to the reference was estimated as the ratio of equally effective doses. The results obtained were compared with assay values reported for in vitro tests - U.S.P. , Howell, colorimetric (Lovibond, Beckman DK-2 spectrophotometer and microelectrophoresis on agarose) . There was no consistent case of one assay methodgiving results greatly different from the others and no assay method consistently described the activity of the heparin preparation in another test system, in vitro or in vivo.

Comparison of Screening Procedures for Mycobacterium tuberculosis Infection Among Patients with Inflammatory Diseases

2009 ◽  
Vol 36 (9) ◽  
pp. 1876-1884 ◽  
Author(s):  
BOLETTE SOBORG ◽  
MORTEN RUHWALD ◽  
MERETE LUND HETLAND ◽  
SØREN JACOBSEN ◽  
AASE BENGAARD ANDERSEN ◽  
...  
Keyword(s):  
Risk Factors ◽  
Mycobacterium Tuberculosis ◽  
Inflammatory Diseases ◽  
Interferon Γ ◽  
Mycobacterium Tuberculosis Infection ◽  
Tb Treatment ◽  
Latent Tb ◽  
Latent Tb Infection ◽  
High Degree

Objective.To test if Mycobacterium tuberculosis screening results differ among patients with inflammatory disease depending on whether the QuantiFeron TB-Gold test (QFT) or tuberculin skin test (TST) is used; and to evaluate if a possible difference is influenced by the presence of risk factors or immunosuppression.Methods.The interferon-γ response to in vitro stimulation of M. tuberculosis-specific antigens was measured with QFT and results were compared with TST. Associations to bacillus Calmette-Guerin (BCG) vaccination, risk factors, and immunosuppression were analyzed for both tests.Results.QFT and TST results were available for 294/302 and 241/302 patients, respectively; 234 had results from both tests. Twenty-one (7%) tested positive with QFT and 45 (19%) with TST. A positive QFT was associated with risk factors for M. tuberculosis infection: i.e., birth or upbringing in a TB-endemic area [risk ratio (RR) = 7.8, 95% CI 1.5–18.2, p < 0.001], previous TB treatment (RR 4.7, 95% CI 1.6–13.5, p = 0.005), and any latent TB infection risk factor (RR 4.7, 95% CI 2.1–11.0, p = 0.0002). Treatment with corticosteroids increased the risk for an inconclusive QFT result (RR 4.2, 95% CI 1.6–10.7, p = 0.04) and decreased the risk for a positive TST result (RR 0.4, 95% CI 0.1–1.0, p = 0.04). Agreement between the tests was low (kappa 0.2, 95% CI 0.02–0.3, p = 0.002).Conclusion.The study documented a high degree of discordant positive QFT and TST results. A positive QFT was more closely associated with risk factors for M. tuberculosis infection than the TST. The use of corticosteroids affected test outcome by increasing the risk for an inconclusive QFT result and decreasing the risk for a positive TST result.

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