Sensitivity of a new commercial enzyme-linked immunospot assay (T SPOT-TB) for diagnosis of tuberculosis in clinical practice

  • T. Meier
  • H.-P. Eulenbruch
  • P. Wrighton-Smith
  • G. Enders
  • T. Regnath
Article

Abstract

Diagnosis of active and latent tuberculosis (TB) remains a challenge; however, over the last few years, a new approach based on detecting Mycobacterium tuberculosis-specific T cells has shown much promise. In particular, there is substantial published evidence showing that the detection of ESAT-6- and CFP-10-specific T cells using the ex vivo enzyme-linked immunospot technique is a marked improvement over the existing tuberculin skin test. This technique, which detects gamma interferon-producing T cells, is now available as the commercial assay T SPOT-TB (Oxford Immunotec, Oxford, UK). In the present study, the usefulness of the T SPOT-TB test for diagnosis of TB in “real-world” clinical practice was investigated. Ninety patients of a southern German referral centre for TB with confirmed or suspected TB were randomly selected for this study. The results of the T SPOT-TB test were compared with the results of conventional diagnostic tools. The T SPOT-TB test detected 70 of 72 patients with pulmonary or extrapulmonary TB, indicating a sensitivity of 97.2% (95% confidence interval, 90.3–99.7). For 45 of these patients, tuberculin skin test (TST) results were also available. Only 40 (89%) of these 45 patients were positive in the TST compared to all 45 (100%) in the T SPOT-TB test (p=0.056). Among 12 of 90 patients for whom active TB disease was ruled out, the T SPOT-TB test was negative for 11 (92%), allowing the rapid exclusion of TB in patients suspected to have active TB disease. The T SPOT-TB test is a sensitive assay for detection of TB and represents a useful addition to the diagnostic algorithm available for detecting TB in low-incidence settings.

Notes

Acknowledgements

The authors are grateful to all patients and the staff of Klinikum Schillerhöhe who took part in this study. We thank Martina Richter for excellent technical assistance performing the T SPOT-TB test. T SPOT-TB reagents were supplied by Oxford Immunotec Limited. We are grateful to Ajit Lalvani for helpful discussions. The experiments performed comply with the current laws of Germany.

References

  1. 1.
    Corbett EL, Watt CJ, Walker N, Maher D, Williams BG, Raviglione MC, Dye C (2003) The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. Arch Intern Med 163:1009–1021CrossRefPubMedGoogle Scholar
  2. 2.
    Jasmer RM, Nahid P, Hopewell PC (2002) Clinical practice. Latent tuberculosis infection. N Engl J Med 347:1860–1866CrossRefPubMedGoogle Scholar
  3. 3.
    Huebner RE, Schein MF, Bass JB Jr (1993) The tuberculin skin test. Clin Infect Dis 17:968–975PubMedGoogle Scholar
  4. 4.
    Delgado JC, Tsai EY, Thim S, Baena A, Boussiotis VA, Reynes JM, Sath S, Grosjean P, Yunis EJ, Goldfeld AE (2002) Antigen-specific and persistent tuberculin anergy in a cohort of pulmonary tuberculosis patients from rural Cambodia. Proc Natl Acad Sci USA 99:7576–7581PubMedGoogle Scholar
  5. 5.
    Wang L, Turner MO, Elwood RK, Schulzer M, FitzGerald JM (2002) A meta-analysis of the effect of Bacille Calmette Guerin vaccination on tuberculin skin test measurements. Thorax 57:804–809CrossRefPubMedGoogle Scholar
  6. 6.
    Behr MA, Wilson MA, Gill WP, Salamon H, Schoolnik GK, Rane S, Small PM (1999) Comparative genomics of BCG vaccines by whole-genome DNA microarray. Science 284:1520–1523CrossRefPubMedGoogle Scholar
  7. 7.
    Andersen P, Munk ME, Pollock JM, Doherty TM (2000) Specific immune-based diagnosis of tuberculosis. Lancet 356:1099–1104CrossRefPubMedGoogle Scholar
  8. 8.
    Pai M, Riley LW, Colford JM Jr (2004) Interferon-gamma assays in the immunodiagnosis of tuberculosis: a systematic review. Lancet Infect Dis 4:761–776CrossRefPubMedGoogle Scholar
  9. 9.
    Dheda K, Udwadia ZF, Huggett JF, Johnson MA, Rook GA (2005) Utility of the antigen-specific interferon-gamma assay for the management of tuberculosis. Curr Opin Pulm Med 11:195–202CrossRefPubMedGoogle Scholar
  10. 10.
    Barnes PF (2001) Diagnosing latent tuberculosis infection: the 100-year upgrade. Am J Respir Crit Care Med 163:807–808PubMedGoogle Scholar
  11. 11.
    Chapman AL, Munkanta M, Wilkinson KA, Pathan AA, Ewer K, Ayles H, Reece WH, Mwinga A, Godfrey-Faussett P, Lalvani A (2002) Rapid detection of active and latent tuberculosis infection in HIV-positive individuals by enumeration of Mycobacterium tuberculosis-specific T cells. AIDS 16:2285–2293PubMedGoogle Scholar
  12. 12.
    Ewer K, Deeks J, Alvarez L, Bryant G, Waller S, Andersen P, Monk P, Lalvani A (2003) Comparison of T-cell-based assay with tuberculin skin test for diagnosis of Mycobacterium tuberculosis infection in a school tuberculosis outbreak. Lancet 361:1168–1173CrossRefPubMedGoogle Scholar
  13. 13.
    Lalvani A, Nagvenkar P, Udwadia Z, Pathan AA, Wilkinson KA, Shastri JS, Ewer K, Hill AV, Mehta A, Rodrigues C (2001) Enumeration of T cells specific for RD1-encoded antigens suggests a high prevalence of latent Mycobacterium tuberculosis infection in healthy urban Indians. J Infect Dis 183:469–477CrossRefPubMedGoogle Scholar
  14. 14.
    Lalvani A, Pathan AA, Durkan H, Wilkinson KA, Whelan A, Deeks JJ, Reece WH, Latif M, Pasvol G, Hill AV (2001) Enhanced contact tracing and spatial tracking of Mycobacterium tuberculosis infection by enumeration of antigen-specific T cells. Lancet 357:2017–2021CrossRefPubMedGoogle Scholar
  15. 15.
    Lalvani A (2003) Spotting latent infection: the path to better tuberculosis control. Thorax 58:916–918CrossRefPubMedGoogle Scholar
  16. 16.
    Richeldi L, Ewer K, Losi M, Hansell DM, Roversi P, Fabbri LM, Lalvani A (2004) Early diagnosis of subclinical multidrug-resistant tuberculosis. Ann Intern Med 140:709–713PubMedGoogle Scholar
  17. 17.
    Richeldi L, Ewer K, Losi M, Bergamini BM, Roversi P, Deeks J, Fabbri LM, Lalvani A (2004) T cell-based tracking of multidrug resistant tuberculosis infection after brief exposure. Am J Respir Crit Care Med 170:288–295CrossRefPubMedGoogle Scholar
  18. 18.
    Lalvani A, Pathan AA, McShane H, Wilkinson RJ, Latif M, Conlon CP, Pasvol G, Hill AV (2001) Rapid detection of Mycobacterium tuberculosis infection by enumeration of antigen-specific T cells. Am J Respir Crit Care Med 163:824–828PubMedGoogle Scholar
  19. 19.
    Ferlinz R (1996) Richtlinien zur Tuberkulindiagnostik. Dtsche Ärztebl 93:1199–1201Google Scholar
  20. 20.
    Russell ND, Hudgens MG, Ha R, Havenar-Daughton C, McElrath MJ (2003) Moving to human immunodeficiency virus type 1 vaccine efficacy trials: defining T cell responses as potential correlates of immunity. J Infect Dis 187:226–242CrossRefPubMedGoogle Scholar
  21. 21.
    Smith JG, Liu X, Kaufhold RM, Clair J, Caulfield MJ (2001) Development and validation of a gamma interferon ELISPOT assay for quantitation of cellular immune responses to varicella-zoster virus. Clin Diagn Lab Immunol 8:871–879CrossRefPubMedGoogle Scholar
  22. 22.
    Pathan AA, Wilkinson KA, Klenerman P, McShane H, Davidson RN, Pasvol G, Hill AV, Lalvani A (2001) Direct ex vivo analysis of antigen-specific IFN-gamma-secreting CD4 T cells in Mycobacterium tuberculosis-infected individuals: associations with clinical disease state and effect of treatment. J Immunol 167:5217–5225PubMedGoogle Scholar
  23. 23.
    Carrara S, Vincenti D, Petrosillo N, Amicosante M, Girardi E, Goletti D (2004) Use of a T cell-based assay for monitoring efficacy of antituberculosis therapy. Clin Infect Dis 38:754–756CrossRefPubMedGoogle Scholar
  24. 24.
    Lalvani A (2004) Counting antigen-specific T cells: a new approach for monitoring response to tuberculosis treatment? Clin Infect Dis 38:757–759CrossRefPubMedGoogle Scholar
  25. 25.
    Liu XQ, Dosanjh D, Varia H, Ewer K, Cockle P, Pasvol G, Lalvani A (2004) Evaluation of T-cell responses to novel RD1- and RD2-encoded Mycobacterium tuberculosis gene products for specific detection of human tuberculosis infection. Infect Immun 72:2574–2581CrossRefPubMedGoogle Scholar
  26. 26.
    Liebeschuetz S, Bamber S, Ewer K, Deeks J, Pathan AA, Lalvani A (2004) Diagnosis of tuberculosis in South African children with a T-cell-based assay: a prospective cohort study. Lancet 364:2196–2203CrossRefPubMedGoogle Scholar
  27. 27.
    Mahairas GG, Sabo PJ, Hickey MJ, Singh DC, Stover CK (1996) Molecular analysis of genetic differences between Mycobacterium bovis BCG and virulent M. bovis. J Bacteriol 178:1274–1282PubMedGoogle Scholar
  28. 28.
    Harboe M, Oettinger T, Wiker HG, Rosenkrands I, Andersen P (1996) Evidence for occurrence of the ESAT-6 protein in Mycobacterium tuberculosis and virulent Mycobacterium bovis and for its absence in Mycobacterium bovis BCG. Infect Immun 64:16–22PubMedGoogle Scholar
  29. 29.
    Arend SM, van Meijgaarden KE, de Boer K, de Palou EC, van Soolingen D, Ottenhoff TH, van Dissel JT (2002) Tuberculin skin testing and in vitro T cell responses to ESAT-6 and culture filtrate protein 10 after infection with Mycobacterium marinum or M. kansasii. J Infect Dis 186:1797–1807CrossRefPubMedGoogle Scholar
  30. 30.
    Pollock JM, Andersen P (1997) The potential of the ESAT-6 antigen secreted by virulent mycobacteria for specific diagnosis of tuberculosis. J Infect Dis 175:1251–1254PubMedGoogle Scholar
  31. 31.
    Lein AD, von Reyn CF, Ravn P, Horsburgh CR Jr, Alexander LN, Andersen P (1999) Cellular immune responses to ESAT-6 discriminate between patients with pulmonary disease due to Mycobacterium avium complex and those with pulmonary disease due to Mycobacterium tuberculosis. Clin Diagn Lab Immunol 6:606–609PubMedGoogle Scholar
  32. 32.
    Doherty TM, Demissie A, Olobo J, Wolday D, Britton S, Eguale T, Ravn P, Andersen P (2002) Immune responses to the Mycobacterium tuberculosis-specific antigen ESAT-6 signal subclinical infection among contacts of tuberculosis patients. J Clin Microbiol 40:704–706CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • T. Meier
    • 1
  • H.-P. Eulenbruch
    • 2
  • P. Wrighton-Smith
    • 3
  • G. Enders
    • 1
  • T. Regnath
    • 1
  1. 1.Medizinisch-Diagnostisches Gemeinschaftslabor Prof. Gisela Enders und PartnerStuttgartGermany
  2. 2.Klinik SchillerhöheLVA WürttembergGerlingenGermany
  3. 3.Oxford Immunotec LimitedAbingdonUK

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