Comparison of an In-house and a Commercial RD1-based ELISPOT-IFN-  Assay for the Diagnosis of Mycobacterium tuberculosis Infection

Mantegani, Paola; Piana, Federica; Codecasa, Luigi; Galli, Laura; Scarpellini, Paolo; Lazzarin, Adriano; Cirillo, Daniela María; Fortis, Claudio

doi:10.3121/cmr.4.4.266

Cited by 15 publications

(12 citation statements)

References 20 publications

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“…TB assays. An in house version of the ELISpot assay has been used in many previous published comparison studies [21, 42–46]. A further limitation of our study was that not every child deemed uninfected had a thorough work-up to exclude TB.…”

Section: Discussionmentioning

confidence: 99%

Detection of tuberculosis in HIV-infected children using an enzyme-linked immunospot assay

Davies

Connell

Johannisen

et al. 2009

Aids

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Objective To evaluate an enzyme-linked immunospot assay (ELISpot) for the diagnosis of tuberculosis (TB) in HIV-infected children with suspected TB and to compare the performance of ELISpot with the tuberculin skin test (TST). Methods IFN-γ responses to Mycobacterium tuberculosis (MTB)-specific antigens were measured by ELISpot in HIV-infected children with suspected TB. HIV-infected and HIV-uninfected children without TB were used for comparison. Results Results were available for 188 children, of which, 139 (74%) were HIV-infected. Of these, 22 were classified as having definite TB, 24 probable TB, 14 possible TB and 128 not having TB. The median (range) age of patients was 20 (10 - 54.1) months. Ninety one percent of ELISpot tests yielded interpretable results. Median IFN-γ responses to early-secreted antigenic target-6 and culture filtrate protein-10 were higher in children with definite or probable TB compared to children without TB (p<0.002). In HIV-infected children with an interpretable result, the ELISpot was positive in 14/21 (66%) with definite TB. A significantly higher proportion of HIV-infected children with definite or probable TB had a positive ELISpot compared to a positive TST (25/39 (64%) vs. 10/34 (29%), p=0.005). In contrast to TST, results from ELISpot were not affected by young age or severe immunosuppression. In HIV-infected children without active TB disease, 27% had a positive ELISpot suggesting latent TB infection. Conclusions ELISpot is more sensitive than TST for the detection of active TB in HIV-infected children. However the sensitivity of current ELISpot assays is not sufficiently high to be used as rule out test for TB.

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Section: Discussionmentioning

confidence: 99%

Detection of tuberculosis in HIV-infected children using an enzyme-linked immunospot assay

Davies

Connell

Johannisen

et al. 2009

Aids

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“…The specificity of TST may be not adequate due to cross-reactions not only with BCG vaccination, but also with non-tuberculous mycobacteria which appears to be rising, especially among young children [ 21 ]. It is important to note that false-positive TST reactions may lead to unnecessary treatment [ 16 , 22 ]. Chemoprophylaxis of TB infected persons is an important component of TB control strategy; however, due to the significant risk of hepatotoxicity and other side effects of anti-TB treatment, chemoprophylaxis cannot be prescribed only on the basis of the TST [ 8 , 10 , 23 ], especially in children.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, we have also analyzed the T SPOT TB responses in relation to TST using different cutoffs, because a TST cutoff ≥5 mm for contacts who have been previously BCG vaccinated is questionable [ 3 , 16 ]. All children with culture-confirmed-TB showed a TST cutoff ≥10 mm.…”

Section: Discussionmentioning

confidence: 99%

“…The responses were considered as positive when the number of spots in the test wells (against at least one of the two tested antigens) was ≥6, if the negative control had 0-5 spots, or ≥2× the number of spots in the negative control wells, if the negative control had 6-10 spots. This cutoff was predefined by the manufacturer's instructions and previous publications [ 15 , 16 ].…”

Section: Methodsmentioning

confidence: 99%

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T-cell-based diagnosis of tuberculosis infection in children in Lithuania: a country of high incidence despite a high coverage with bacille Calmette-Guerin vaccination

Hansted

Andriuškevičienė

Sakalauskas³

et al. 2009

BMC Pulm Med

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BackgroundLithuania is a country with a high incidence of tuberculosis (TB), despite a high coverage with bacille Calmette-Guerin (BCG) vaccination. Until now the only method used to detect latent TB infection was the tuberculin skin test (TST). However, TST may have a cross reactivity to the BCG vaccine and to environmental mycobacteria. The aim of this study was to conduct assessments of the diagnostic accuracy of the T-cell based test (T SPOT TB) for TB in children who had previously been BCG vaccinated and compare these with the results of the TST.MethodsBetween January 2005 and February 2007, children with bacteriologically confirmed TB, children having contacts with a case of infectious pulmonary TB and children without any known risk for TB were tested with both the TST and T SPOT TB.ResultsThe TST and T SPOT TB tests were positive for all patients in the „culture-confirmed TB“ group. Whereas, in the „high risk for TB“ group, the TST was positive for 60%, but the T SPOT TB test, only for 17.8%. Meanwhile the results for the „low risk for TB“ group were 65.4% and 9.6%, respectively. A correlation between the TST and T SPOT TB was obtained in the "culture-confirmed TB" group where the TST ≥15 mm (r = 0.35, p < 0.001).ConclusionThe T-cell based method is more objective than the TST for identifying latent TB infection in children who had been previously BCG vaccinated. This method could be useful in countries like Lithuania where there is a high incidence of TB despite a high coverage with BCG vaccination. It may also help to avoid unnecessary chemoprophylaxis when TST reactions are false-positive.

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“…During the past several years, a large number of studies based on CMI responses have been performed to identify mycobacterial antigens of diagnostic potential [18][19][20][21][22] . These studies have shown that secreted antigens of M. tuberculosis are the primary targets of cellular immune responses in TB patients.…”

Section: Discussionmentioning

confidence: 99%

Cell-Mediated Immune Responses to Complex and Single Mycobacterial Antigens in Tuberculosis Patients with Diabetes

et al. 2008

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Objective: To evaluate cell-mediated immune (CMI) response in diabetic and non-diabetic tuberculosis (TB) patients and healthy subjects in response to complex, fractionated and single antigens of Mycobacteriumtuberculosis. Material and Methods: Peripheral blood mononuclear cells (PBMC) were obtained from patients suffering from pulmonary TB and type II diabetes (n = 7), pulmonary TB without diabetes (n = 10) and healthy subjects without TB and diabetes (n = 10). PBMC were assessed for CMI responses in antigen-induced proliferation assays in response to complex mycobacterial antigens (whole cells, cell walls and culture filtrate of M. tuberculosis), a battery of naturally purified or recombinant produced secreted (ESAT6, MPT59, MPT64 and MTB38) and cytosolic (MTB10, MTB70, ML10, ML28, ML36, ML65 and MB65) mycobacterial antigens and fractionated culture filtrate proteins (fractions F1–F10) of M. tuberculosis. Results:The majority (>70%) of diabetic and non-diabetic TB patients and healthy subjects responded to the complex antigens of M. tuberculosis. However, among the single antigens, ESAT6 was most frequently recognized by TB patients with and without diabetes, but least recognized by healthy subjects. The secreted antigens MPT59 and MPT64 were recognized by all the groups, whereas the cytosolic antigens were recognized best by healthy subjects. When tested with fractionated secreted proteins present in the culture filtrate of M. tuberculosis, the best responses in both diabetic and non-diabetic TB patients were obtained with fractions containing low-molecular-weight proteins. Conclusions: Diabetic and non-diabetic TB patients respond frequently to secreted low-molecular-weight ESAT6 antigen of M. tuberculosis, indicating that this antigen may be useful in the diagnosis of TB in both the groups.

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Comparison of an In-house and a Commercial RD1-based ELISPOT-IFN- Assay for the Diagnosis of Mycobacterium tuberculosis Infection

Cited by 15 publications

References 20 publications

Detection of tuberculosis in HIV-infected children using an enzyme-linked immunospot assay

Detection of tuberculosis in HIV-infected children using an enzyme-linked immunospot assay

T-cell-based diagnosis of tuberculosis infection in children in Lithuania: a country of high incidence despite a high coverage with bacille Calmette-Guerin vaccination

Cell-Mediated Immune Responses to Complex and Single Mycobacterial Antigens in Tuberculosis Patients with Diabetes

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