Abstract
Longitudinal studies regarding the reproducibility of Interferon-gamma release assay (IGRA) T-SPOT.TB for the diagnosis of Mycobacterium tuberculosis (M. tb) infection in serial testing are limited. We retrospectively analysed results of serially tested subjects in a medical laboratory in Germany over a time period of 14 years. From October 2004 to December 2018, a total of 5440 subjects were identified with a second T-SPOT.TB test after a median time interval of 258 days (interquartile range [IQR] 62–665). Consistently negative (n = 4520) or positive results (n = 682) were observed in 5202 (95.6%) subjects, indicating a high degree of concordance in serial testing (κ = 0.83). Test conversions occurred in 101 of 4621 (2.2%) subjects with initially negative tests. Of 819 subjects with initially positive test results, 137 (16.7%) had a test reversion which was associated with low spot numbers of the first test. Of 529 subjects retested within 1 year, only 60 (11.3%) displayed a test reversion. In subjects retested after more than 1 year, 77 of 290 (26.6%) tests reverted. This significantly higher rate of test reversions after more than 1 year was age-dependent and only observed in subjects above the age of 40 years. In the medical laboratory, the T-SPOT.TB test demonstrates a high reproducibility in serial testing.
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Data availability
The data that support the findings of this study are available from the corresponding author [TM] upon reasonable request.
References
Lalvani A, Pareek M (2010) A 100 year update on diagnosis of tuberculosis infection. Br Med Bull 93:69–84. https://doi.org/10.1093/bmb/ldp039
Pai M, Zwerling A, Menzies D (2008) Systematic review: t-cell-based assays for the diagnosis of latent tuberculosis infection: an update. Ann Intern Med 149:177–184. https://doi.org/10.7326/0003-4819-149-3-200808050-00241
Getahun H, Matteelli A, Abubakar I, Aziz MA, Baddeley A, Barreira D et al (2015) Management of latent Mycobacterium tuberculosis infection: WHO guidelines for low tuberculosis burden countries. Eur Respir J 46:1563–1576. https://doi.org/10.1183/13993003.01245-2015
European Centre for Disease Prevention and Control, editor (2011) Use of interferon-gamma release assays in support of TB diagnosis: ad hoc scientific panel opinion. [Luxembourg: Publ. Off.]. https://doi.org/10.2900/38588
Ringshausen FC, Schablon A, Nienhaus A. Interferon-gamma release assays for the tuberculosis serial testing of health care workers: a systematic review. Journal of occupational medicine and toxicology (London, England) 2012 26;7:6. doi: https://doi.org/10.1186/1745-6673-7-6
Tagmouti S, Slater M, Benedetti A, Kik SV, Banaei N, Cattamanchi A, et al. Reproducibility of interferon gamma (IFN-γ) release assays. A systematic review. Ann Am Thorac Soc 2014;11:1267–1276. doi: https://doi.org/10.1513/AnnalsATS.201405-188OC
Banaei N, Gaur RL, Pai M (2016) Interferon gamma release assays for latent tuberculosis: what are the sources of variability? J Clin Microbiol 54:845–850. https://doi.org/10.1128/JCM.02803-15
Wilkinson KA, Kon OM, Newton SM, Meintjes G, Davidson RN, Pasvol G, et al. (2006) Effect of treatment of latent tuberculosis infection on the T cell response to Mycobacterium tuberculosis antigens. J Infect Dis 1(193):354–9. https://doi.org/10.1086/499311
van Zyl-Smit RN, Zwerling A, Dheda K, Pai M (2009) Within-subject variability of interferon-g assay results for tuberculosis and boosting effect of tuberculin skin testing: a systematic review. PLoS One 4:e8517. https://doi.org/10.1371/journal.pone.0008517
Lewinsohn DM, Leonard MK, LoBue PA, Cohn DL, Daley CL, Desmond E et al (2017) Official American thoracic society/infectious diseases society of America/Centers for disease control and prevention clinical practice guidelines: diagnosis of tuberculosis in adults and children. Clin Infect Dis 64:111–115. https://doi.org/10.1093/cid/ciw778
Landis JR, Koch GG (1977) An application of hierarchical kappa-type statistics in the assessment of majority agreement among multiple observers. Biometrics 33:363–374
King TC, Upfal M, Gottlieb A, Adamo P, Bernacki E, Kadlecek CP et al (2015) T-SPOT.TB interferon-γ release assay performance in healthcare worker screening at nineteen U.S. hospitals. Am J Respir Crit Care Med 192:367–373. https://doi.org/10.1164/rccm.201501-0199OC
Teranishi S, Kobayashi N, Aoki A, Katakura S, Yamamoto M, Koizumi H et al (2019) Reproducibility of the T-SPOT.TB test for screening Mycobacterium tuberculosis infection in Japan. J Infect Chemother. https://doi.org/10.1016/j.jiac.2019.08.006
Dorman SE, Belknap R, Graviss EA, Reves R, Schluger N, Weinfurter P et al (2014) Interferon-γ release assays and tuberculin skin testing for diagnosis of latent tuberculosis infection in healthcare workers in the United States. Am J Respir Crit Care Med 189:77–87. https://doi.org/10.1164/rccm.201302-0365OC
Janetzki S, Schaed S, Blachere NEB, Ben-Porat L, Houghton AN, Panageas KS (2004) Evaluation of Elispot assays: influence of method and operator on variability of results. J Immunol Methods 291:175–183. https://doi.org/10.1016/j.jim.2004.06.008
Janetzki S, Panageas KS, Ben-Porat L, Boyer J, Britten CM, Clay TM, et al. Results and harmonization guidelines from two large-scale international Elispot proficiency panels conducted by the Cancer Vaccine Consortium (CVC/SVI). Cancer Immunol Immunother 2008;57:303–315. doi: https://doi.org/10.1007/s00262-007-0380-6
Tuuminen T, Tavast E, Väisänen R, Himberg J-J, Seppälä I. Assessment of imprecision in gamma interferon release assays for the detection of exposure to Mycobacterium tuberculosis. Clin Vaccine Immunol 2010;17:596–601. doi: https://doi.org/10.1128/CVI.00320-09
Oxford Immunotec. T-SPOT.TB test 96-Well Plate Format (TB.200) PACKAGE INSERT [Internet]. 2009 [cited 2012 1]; http://www.oxfordimmunotec.com
FDA, U.S. Food and Drug Administration. PM A P070006: FDA summary of safety and effectiveness data [Internet]. FDA, U.S. Food and Drug Administration; 2008. http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cftopic/pma/pma.cfm?num=p070006
Rego K, Pereira K, MacDougall J, Cruikshank W (2018) Utility of the T-SPOT®.TB test’s borderline category to increase test resolution for results around the cut-off point. Tuberculosis (Edinb) 108:178–185. https://doi.org/10.1016/j.tube.2017.12.005
Zhang H-C, Ruan Q-L, Wu J, Zhang S, Yu S-L, Wang S, et al. Serial T-SPOT.TB in household contacts of tuberculosis patients: a 6-year observational study in China. Int. J Tuberc Lung Dis 2019;23:989–995. doi: https://doi.org/10.5588/ijtld.18.0252
Ewer K, Millington KA, Deeks JJ, Alvarez L, Bryant G, Lalvani A (2006) Dynamic antigen-specific T-cell responses after point-source exposure to Mycobacterium tuberculosis. Am J Respir Crit Care Med 174:831–839. https://doi.org/10.1164/rccm.200511-1783OC
Dutta NK, Karakousis PC (2014) Latent tuberculosis infection: myths, models, and molecular mechanisms. Microbiol Mol Biol Rev 78:343–371. https://doi.org/10.1128/MMBR.00010-14
Clifford V, He Y, Zufferey C, Connell T, Curtis N (2015) Interferon gamma release assays for monitoring the response to treatment for tuberculosis: a systematic review. Tuberculosis (Edinb) 95:639–650. https://doi.org/10.1016/j.tube.2015.07.002
Perry S, Parsonnet J (2005) Commentary: H. pylori infection in early life and the problem of imperfect tests. Int J Epidemiol 34:1356–1358. https://doi.org/10.1093/ije/dyi243
Perry S, Sanchez L, Yang S, Agarwal Z, Hurst P, Parsonnet J (2008) Reproducibility of QuantiFERON-TB gold in-tube assay. Clin Vaccine Immunol 15:425–432. https://doi.org/10.1128/CVI.00398-07
Brodhun B, Altmann D, Hauer B, Haas W. Bericht zur Epidemiologie der Tuberkulose in Deutschland für 2017. 2018; www.rki.de/tuberkulosedoi:https://doi.org/10.17886/rkipubl-2018-002
Bienek DR, Chang CK (2009) Evaluation of an interferon-gamma release assay, T-SPOT.TB, in a population with a low prevalence of tuberculosis. Int J Tuberc Lung Dis 13:1416–1421
Talbot EA, Harland D, Wieland-Alter W, Burrer S, Adams LV (2012) Specificity of the tuberculin skin test and the T-SPOT.TB assay among students in a low-tuberculosis incidence setting. J Am Coll Heal 60:94–96. https://doi.org/10.1080/07448481.2011.580029
Meier T, Eulenbruch H-P, Wrighton-Smith P, Enders G, Regnath T (2005) Sensitivity of a new commercial enzyme-linked immunospot assay (T SPOT-TB) for diagnosis of tuberculosis in clinical practice. Eur J Clin Microbiol Infect Dis 24:529–536. https://doi.org/10.1007/s10096-005-1377-8
Linton PJ, Dorshkind K (2004) Age-related changes in lymphocyte development and function. Nat Immunol 5:133–139. https://doi.org/10.1038/ni1033
Bae W, Park KU, Song EY, Kim SJ, Lee YJ, Park JS, et al. Comparison of the sensitivity of QuantiFERON-TB gold in-tube and T-SPOT.TB according to patient age. PLoS One 2016;11:e0156917. doi: https://doi.org/10.1371/journal.pone.0156917
Johnston RN, Ritchie RT, Murray IH (1963) Declining tuberculin sensitivity with advancing age. Br Med J 21(2):720–724. https://doi.org/10.1136/bmj.2.5359.720
Grzybowski S, Allen EA (1964) The challenge of tuberculosis in decline. A study based on the epidemiology of tuberculosis in Ontario, Canada. Am Rev Respir Dis 90:707–720. https://doi.org/10.1164/arrd.1964.90.5.707
Karam F, Mbow F, Fletcher H, Senghor CS, Coulibaly KD, LeFevre AM et al (2008) Sensitivity of IFN-gamma release assay to detect latent tuberculosis infection is retained in HIV-infected patients but dependent on HIV/AIDS progression. PLoS One 3:e1441. https://doi.org/10.1371/journal.pone.0001441
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TM designed the study, performed data and statistical analysis. Both authors contributed to writing the manuscript.
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TM had received honoraria from Oxford Immunotec for speaking at symposia and teaching practical T-SPOT.TB test processing. ME does not have any conflict of interest. Oxford Immunotec had no role in study design, data collection and analysis, decision to publish or manuscript preparation.
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The Ethics Committee of the Landesärztekammer Baden-Württemberg approved the study (reference F-2019-059).
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Appendix
Appendix
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Expected conversions % =(1 − NPV) × SE + NPV × (1 − SP) [26]
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Expected reversions % =PPV × (1 − SE) + (1 − PPV) × SP [26]
$$ {\displaystyle \begin{array}{c}\mathrm{NPV}=\frac{\mathrm{SP}\times \left(1-\mathrm{PRE}\right)}{\mathrm{SP}\times \left(1-\mathrm{PRE}\right)+\left(1-\mathrm{SE}\right)\times \mathrm{PRE}}\\ {}\mathrm{PPV}=\frac{\mathrm{PRE}\times \mathrm{SE}}{\mathrm{PRE}\times \mathrm{SE}+\left(1-\mathrm{PRE}\right)\times \left(1-\mathrm{SP}\right)}\end{array}} $$
where NPV, PPV, SE, SP and PRE are the negative predictive value, positive predictive value, sensitivity, specificity and prevalence of TB infection, respectively.
The rate of 15.1% initially positive subjects is assumed to reflect the prevalence of TB infection in our study population.
Calculation example for the observed frequency of 1.3% conversions:
For sensitivity = 0.970, the specificity is 0.992.
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Meier, T., Enders, M. High reproducibility of the interferon-gamma release assay T-SPOT.TB in serial testing. Eur J Clin Microbiol Infect Dis 40, 85–93 (2021). https://doi.org/10.1007/s10096-020-03997-3
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DOI: https://doi.org/10.1007/s10096-020-03997-3