Screening for tuberculosis infections (TBI) using the tuberculin skin test or interferon-gamma release assays (IGRA) is crucial in controlling the global TB burden. This study evaluates the performance of a new IGRA for the detection of T-cell responses against Mycobacterium tuberculosis. Blood samples from 34 adults with tuberculosis disease (TB) and from 30 children with TB, TBI or without TB were analyzed using the prototype Quan-T-Cell TB (EUROIMMUN). The pediatric samples were additionally measured using the established QuantiFERON-TB Gold Plus assay (Qiagen). Clinical performance and inter-assay concordance were analyzed. The prototype Quan-T-Cell TB yielded positivity rates of 88.2% and 100% in adults with TB and children with TBI, respectively, at a specificity of 93.8%. Comparison between the two IGRAs showed positive, negative and overall agreement rates of 100%, 93.8% and 96.3%, respectively, with a kappa score of 0.924 indicating almost perfect agreement. Our study shows promising results of the new prototype Quan-T-Cell TB, as reflected by high concordance with the final diagnosis in adults and children and performance comparable to that of the QuantiFERON IGRA. In individual cases, the data suggest that the prototype Quan-T-Cell TB may be even more consistent with TBI-related clinical findings. Unlike the QuantiFERON assay, the Quan-T-Cell TB has a predefined borderline range, which is advantageous as it may help to differentiate non-specific variation near the cut-off, and fewer sample tubes are required per analysis. The new Quan-T-Cell TB may therefore be a good alternative to the established QuantiFERON IGRA for TBI screening. Further assay optimization is underway, including evaluation studies based on larger patient and control cohorts.

Department of Pediatrics 1st Faculty of Medicine Charles University and Thomayer University Hospital Prague Czech Republic

Department of Respiratory Medicine 1st Faculty of Medicine Charles University and Thomayer University Hospital Prague Czech Republic

Institute for Experimental Immunology affiliated to EUROIMMUN Medizinische Labordiagnostika AG Lübeck Germany

Laboratory of Immunology Thomayer University Hospital Prague Czech Republic

Zobrazit více v PubMed

Andersen P, Munk ME, Pollock JM, Doherty TM (2000) Specific immune-based diagnosis of tuberculosis. Lancet 356(9235):1099–1104. 10.1016/S0140-6736(00)02742-2 PubMed

Behr MA, Kaufmann E, Duffin J, Edelstein PH, Ramakrishnan L (2021) Latent tuberculosis: two centuries of confusion. Am J Respir Crit Care Med 204(2):142–148. 10.1164/rccm.202011-4239PP PubMed PMC

Buonsenso D, Delogu G, Perricone C, Grossi R, Careddu A, De MF, Palucci I, Sanguinetti M, Valentini P, Sali M (2020) Accuracy of QuantiFERON-TB Gold Plus Test for diagnosis of Mycobacterium tuberculosis infection in children. J Clin Microbiol 58(6):e00272-20. 10.1128/jcm.00272-20 PubMed PMC

Buonsenso D, Seddon JA, Esposito S, Barcellini L (2023) QuantiFERON-TB Gold Plus performance in children: a narrative review. Pediatr Infect Dis J 42(5):e158-165. 10.1097/INF.0000000000003877 PubMed PMC

Centers for Disease Control and Prevention (CDC) (2020) Tuberculin Skin Testing Fact Sheet. Available online:https://www.cdc.gov/tb/publications/factsheets/testing/Tuberculin_Skin_Testing_Information_for_Health_Care_Providers.pdf. Accessed 19 Nov 2024

Cohen A, Mathiasen VD, Schön T, Wejse C (2019) The global prevalence of latent tuberculosis: a systematic review and meta-analysis. Eur Respir J 54(3):1900655. 10.1183/13993003.00655-2019 PubMed

Cole B, Nilsen DM, Will L, Etkind SC, Burgos M, Chorba T (2020) Essential Components of a public health tuberculosis prevention, control, and elimination program: recommendations of the advisory council for the elimination of tuberculosis and the national tuberculosis controllers association. MMWR Recomm Rep 69(7):1–27. 10.15585/mmwr.rr6907a1 PubMed PMC

Doležalová K, Hadlova P, Ibrahimova M, Golias J, Baca L, Kopecka E, Sukholytka M, Koziar Vasakova M (2024a) Flow cytometry-based method using diversity of cytokine production differentiates between Mycobacterium tuberculosis infection and disease. Tuberculosis (Edinb) 147. 10.1016/j.tube.2024.102518 PubMed

Doležalová K, Koziar Vašáková M, Pohunek P (2024b) New recommendations in the care of pediatric and adolescent patients with tuberculosis. Official opinion of the Children’s Pulmonology Society ÈLS JEP and the Society for Pneumology and Phthisiology ÈLS JEP. Ces slov Pediat 79(Suppl 1):S25–35. 10.55095/CSPediatrie2024/031

European Centre for Disease Prevention and Control (ECDC) (2011) Use of interferon-gamma release assays in support of TB diagnosis. Available online: https://www.ecdc.europa.eu/sites/default/files/media/en/publications/Publications/1103_GUI_IGRA.pdf. Accessed 19 Nov 2024

Farhat M, Greenaway C, Pai M, Menzies D (2006) False-positive tuberculin skin tests: what is the absolute effect of BCG and non-tuberculous mycobacteria? Int J Tuberc Lung Dis 10(11):1192–1204. https://www.ingentaconnect.com/content/iuatld/ijtld/2006/00000010/00000011/art00003. Accessed 19 Nov 2024 PubMed

Ferrara G, Losi M, Meacci M, Meccugni B, Piro R, Roversi P, Bergamini BM, D’Amico R, Marchegiano P, Rumpianesi F, Fabbri LM, Richeldi L (2005) Routine hospital use of a new commercial whole blood interferon-gamma assay for the diagnosis of tuberculosis infection. Am J Respir Crit Care Med 172(5):631–635. 10.1164/rccm.200502-196OC PubMed

Fox A, Jeffries DJ, Hill PC, Hammond AS, Lugos MD, Jackson-Sillah D, Donkor SA, Owiafe PK, McAdam KP, Brookes RH (2007) ESAT-6 and CFP-10 can be combined to reduce the cost of testing for Mycobacterium tuberculosis infection, but CFP-10 responses associate with active disease. Trans R Soc Trop Med Hyg 101(7):691–698. 10.1016/j.trstmh.2007.03.001 PubMed

Getahun H, Matteelli A, Abubakar I, Aziz MA, Baddeley A, Barreira D, Den BS, Borroto Gutierrez SM, Bruchfeld J, Burhan E, Cavalcante S, Cedillos R, Chaisson R, Chee CB, Chesire L et al (2015) Management of latent Mycobacterium tuberculosis infection: WHO guidelines for low tuberculosis burden countries. Eur Respir J 46(6):1563–1576. 10.1183/13993003.01245-2015 PubMed PMC

Gopalaswamy R, Dusthackeer VN, Kannayan S, Subbian S (2021) Extrapulmonary tuberculosis—an update on the diagnosis, treatment and drug resistance. J Respir 1(2):141–164. 10.3390/jor1020015

Greveson K, Murray C, Toumpanakis C, Epstein O, Hamilton M, Lipman M (2011) Interferon gamma-release assays for detecting latent tuberculosis infection in patients scheduled for anti-TNFalpha therapy. Frontline Gastroenterol 2(1):26–31. 10.1136/fg.2010.000356 PubMed PMC

Hamada Y, Den BS, Cirillo DM, Penn-Nicholson A, Ruhwald M, Menzies D, Oxlade O, Falzon D, Kanchar A, Korobitsyn A, Zignol M, Matteelli A, Kasaeva T (2021) Framework for the evaluation of new tests for tuberculosis infection. Eur Respir J 58(2):2004078. 10.1183/13993003.04078-2020 PubMed PMC

Herrera V, Perry S, Parsonnet J, Banaei N (2011) Clinical application and limitations of interferon-gamma release assays for the diagnosis of latent tuberculosis infection. Clin Infect Dis 52(8):1031–1037. 10.1093/cid/cir068 PubMed

Huang CT, Ruan SY, Tsai YJ, Kuo PH, Ku SC, Lee PL, Kuo LC, Hsu CL, Huang CK, Yang CY, Chien YC, Wang JY, Yu CJ (2016) Effects of acute critical illnesses on the performance of interferon-gamma release assay. Sci Rep 6:19972. 10.1038/srep19972 PubMed PMC

Infantino M, Tsalouchos A, Russo E, Laudicina S, Grossi V, Lari B, Benucci M, Stacchini L, Amedei A, Casprini P, Villalta D, Dattolo PC, Manfredi M (2022) Assessing T-cell immunity in kidney transplant recipients with absent antibody production after a 3rd dose of the mRNA-1273 vaccine. Int J Mol Sci 23(20):12333. 10.3390/ijms232012333 PubMed PMC

Kim WS, Choi JI, Cheon JE, Kim IO, Yeon KM, Lee HJ (2006) Pulmonary tuberculosis in infants: radiographic and CT findings. AJR Am J Roentgenol 187(4):1024–1033. 10.2214/AJR.04.0751 PubMed

Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33(1):159–174. 10.2307/2529310 PubMed

Latorre I, de Souza-Galvao M, Ruiz-Manzano J, Lacoma A, Prat C, Altet N, Ausina V, Dominguez J (2010) Evaluating the non-tuberculous mycobacteria effect in the tuberculosis infection diagnosis. Eur Respir J 35(2):338–342. 10.1183/09031936.00196608 PubMed

Latorre I, Tytarenko N, Díaz S, Zapf D, Grage-Griebenow E, Saschenbrecker S, de Souza-Galvao M, Casas X, Molina I, Forcada N, Santiago J, Stojanovic Z, Domínguez J (2024) Detection of T-cell responses in tuberculosis patients using a new prototype interferon-gamma release assay. ESCMID Global Abstract Book 2024: 34th Congress of the European societey of clinical microbiology and infectious diseases, Barcelona, Spain, 27-30 April 2024. CMI Communications 1(Suppl):100013, Abstract P0640|03042. 10.1016/j.cmicom.2024.100013

Lewinsohn DM, Leonard MK, LoBue PA, Cohn DL, Daley CL, Desmond E, Keane J, Lewinsohn DA, Loeffler AM, Mazurek GH, O’Brien RJ, Pai M, Richeldi L, Salfinger M, Shinnick TM 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(2):111–115. 10.1093/cid/ciw778 PubMed PMC

Lin PL, Flynn JL (2018) The end of the binary era: revisiting the spectrum of tuberculosis. J Immunol 201(9):2541–2548. 10.4049/jimmunol.1800993 PubMed PMC

Lin SY, Desmond EP (2014) Molecular diagnosis of tuberculosis and drug resistance. Clin Lab Med 34(2):297–314. 10.1016/j.cll.2014.02.005 PubMed

Mancuso JD, Mody RM, Olsen CH, Harrison LH, Santosham M, Aronson NE (2017) The long-term effect of Bacille Calmette-Guerin vaccination on tuberculin skin testing: a 55-year follow-up study. Chest 152(2):282–294. 10.1016/j.chest.2017.01.001 PubMed

Mazurek GH, Jereb J, Vernon A, LoBue P, Goldberg S, Castro K (2010) Updated guidelines for using interferon gamma release assays to detect mycobacterium tuberculosis infection - United States, 2010. MMWR Recomm Rep 59(RR05):1–25.https://www.cdc.gov/mmwr/preview/mmwrhtml/rr5905a1.htm . Accessed 19 Nov 2024 PubMed

Mazurek GH, LoBue PA, Daley CL, Bernardo J, Lardizabal AA, Bishai WR, Iademarco MF, Rothel JS (2001) Comparison of a whole-blood interferon gamma assay with tuberculin skin testing for detecting latent Mycobacterium tuberculosis infection. JAMA 286(14):1740–1747. 10.1001/jama.286.14.1740 PubMed

Meier NR, Volken T, Geiger M, Heininger U, Tebruegge M, Ritz N (2019) Risk factors for indeterminate interferon-gamma release assay for the diagnosis of tuberculosis in children-a systematic review and meta-analysis. Front Pediatr 7:208. 10.3389/fped.2019.00208 PubMed PMC

Metcalfe JZ, Cattamanchi A, McCulloch CE, Lew JD, Ha NP, Graviss EA (2013) Test variability of the QuantiFERON-TB gold in-tube assay in clinical practice. Am J Respir Crit Care Med 187(2):206–211. 10.1164/rccm.201203-0430OC PubMed PMC

Nemes E, Rozot V, Geldenhuys H, Bilek N, Mabwe S, Abrahams D, Makhethe L, Erasmus M, Keyser A, Toefy A, Cloete Y, Ratangee F, Blauenfeldt T, Ruhwald M, Walzl G et al (2017) Optimization and interpretation of serial QuantiFERON testing to measure acquisition of Mycobacterium tuberculosis infection. Am J Respir Crit Care Med 196(5):638–648. 10.1164/rccm.201704-0817OC PubMed PMC

Nolt D, Starke JR (2021) Tuberculosis infection in children and adolescents: testing and treatment. Pediatrics 148(6):e2021054663. 10.1542/peds.2021-054663 PubMed

Pai M, Denkinger CM, Kik SV, Rangaka MX, Zwerling A, Oxlade O, Metcalfe JZ, Cattamanchi A, Dowdy DW, Dheda K, Banaei N (2014) Gamma interferon release assays for detection of Mycobacterium tuberculosis infection. Clin Microbiol Rev 27(1):3–20. 10.1128/cmr.00034-13 PubMed PMC

Pai M, Menzies D (2007) Interferon-gamma release assays: what is their role in the diagnosis of active tuberculosis? Clin Infect Dis 44(1):74–77. 10.1086/509927 PubMed

Petruccioli E, Vanini V, Chiacchio T, Cuzzi G, Cirillo DM, Palmieri F, Ippolito G, Goletti D (2017) Analytical evaluation of QuantiFERON- Plus and QuantiFERON- Gold In-tube assays in subjects with or without tuberculosis. Tuberculosis (Edinb) 106:38–43. 10.1016/j.tube.2017.06.002 PubMed

Qiagen (2024a) QuantiFERON-TB Gold Plus (QFT-Plus) blood collection tubes instructions for use. Available online: https://www.qiagen.com/us/resources/resourcedetail?id=efe732d5-4b93-477f-983b-7390d0a14637&lang=en. Accessed 19 Nov 2024

Qiagen (2024b) QuantiFERON-TB Gold Plus (QFT-Plus) ELISA package insert. Available online: https://www.qiagen.com/us/resources/resourcedetail?id=ac068fc7-a994-4443-ac7c-dda43ce2bc5e&lang=en. Accessed 19 Nov 2024

Redelman-Sidi G, Sepkowitz KA (2013) IFN-gamma release assays in the diagnosis of latent tuberculosis infection among immunocompromised adults. Am J Respir Crit Care Med 188(4):422–431. 10.1164/rccm.201209-1621CI PubMed PMC

Santoro-Lopes G, Subramanian AK, Molina I, Aguado JM, Rabagliatti R, Len O (2018) Tuberculosis recommendations for solid organ transplant recipients and donors. Transplantation 102(Suppl 2):S60–S65. 10.1097/TP.0000000000002014 PubMed

Santos JA, Durate R, Nunes C (2020) Host factors associated to false negative and indeterminate results in an interferon-gamma release assay in patients with active tuberculosis. Pulmonology 26(6):353–362. 10.1016/j.pulmoe.2019.11.001 PubMed

Schablon A, Nienhaus A, Ringshausen FC, Preisser AM, Peters C (2014) Occupational screening for tuberculosis and the use of a borderline zone for interpretation of the IGRA in German healthcare workers. PLoS One 9(12):e115322. 10.1371/journal.pone.0115322 PubMed PMC

Selwyn PA, Hartel D, Lewis VA, Schoenbaum EE, Vermund SH, Klein RS, Walker AT, Friedland GH (1989) A prospective study of the risk of tuberculosis among intravenous drug users with human immunodeficiency virus infection. N Engl J Med 320(9):545–550. 10.1056/NEJM198903023200901 PubMed

Sester M, Sotgiu G, Lange C, Giehl C, Girardi E, Migliori GB, Bossink A, Dheda K, Diel R, Dominguez J, Lipman M, Nemeth J, Ravn P, Winkler S, Huitric E et al (2011) Interferon-gamma release assays for the diagnosis of active tuberculosis: a systematic review and meta-analysis. Eur Respir J 37(1):100–111. 10.1183/09031936.00114810 PubMed

Shafeque A, Bigio J, Hogan CA, Pai M, Banaei N (2020) Fourth-generation QuantiFERON-TB Gold Plus: what is the evidence? J Clin Microbiol 58(9):e01950-19. 10.1128/jcm.01950-19 PubMed PMC

Soler-Garcia A, Gamell A, Perez-Porcuna T, Soriano-Arandes A, Santiago B, Tortola T, Ruiz-Serrano MJ, Korta Murua JJ, Bustillo-Alonso M, Garrote-Llanos MI, Rodriguez-Molino P, Piqueras AI, Tagarro A, Monsonis M, Tebruegge M et al (2022) Performance of QuantiFERON-TB Gold Plus assays in children and adolescents at risk of tuberculosis: a cross-sectional multicentre study. Thorax 77(12):1193–1201. 10.1136/thoraxjnl-2021-217592 PubMed

Tagmouti S, Slater M, Benedetti A, Kik SV, Banaei N, Cattamanchi A, Metcalfe J, Dowdy D, van Zyl SR, Dendukuri N, Pai M, Denkinger C (2014) Reproducibility of interferon gamma (IFN-gamma) release assays. A systematic review. Ann Am Thorac Soc 11(8):1267–1276. 10.1513/AnnalsATS.201405-188OC PubMed PMC

Theel ES, Hilgart H, Breen-Lyles M, McCoy K, Flury R, Breeher LE, Wilson J, Sia IG, Whitaker JA, Clain J, Aksamit TR, Escalante P (2018) Comparison of the QuantiFERON-TB Gold Plus and QuantiFERON-TB Gold In-Tube interferon gamma release assays in patients at risk for tuberculosis and in health care workers. J Clin Microbiol 56(7):e00614-18. 10.1128/jcm.00614-18 PubMed PMC

van Zyl-Smit RN, Pai M, Peprah K, Meldau R, Kieck J, Juritz J, Badri M, Zumla A, Sechi LA, Bateman ED, Dheda K (2009) Within-subject variability and boosting of T-cell interferon-gamma responses after tuberculin skin testing. Am J Respir Crit Care Med 180(1):49–58. 10.1164/rccm.200811-1704OC PubMed

Vašáková M (2013) Tuberculosis in the Czech Republic. Current status. Diagnosis, treatment and prevention. Vnitr Lek 59(4):284–289. https://pubmed.ncbi.nlm.nih.gov/23711054/. Accessed 19 Nov 2024 PubMed

Venkatappa TK, Punnoose R, Katz DJ, Higgins MP, Banaei N, Graviss EA, Belknap RW, Ho CS (2019) Comparing QuantiFERON-TB Gold Plus with other tests to diagnose Mycobacterium tuberculosis infection. J Clin Microbiol 57(11):e00985-19. 10.1128/jcm.00985-19 PubMed PMC

Wikell A, Jonsson J, Dyrdak R, Henningsson AJ, Eringfalt A, Kjerstadius T, Halldin E, Baqir H, Kholod V, Sturegard E, Bruchfeld J, Schon T (2021) The impact of borderline Quantiferon-TB Gold Plus results for latent tuberculosis screening under routine conditions in a low-endemicity setting. J Clin Microbiol 59(12):e0137021. 10.1128/jcm.01370-21 PubMed PMC

World Health Organization (WHO) (2023) WHO Global Tuberculosis Report 2023. Available online: https://www.who.int/teams/global-tuberculosis-programme/tb-reports/global-tuberculosis-report-2023. Accessed 19 Nov 2024

Yi L, Sasaki Y, Nagai H, Ishikawa S, Takamori M, Sakashita K, Saito T, Fukushima K, Igarashi Y, Aono A, Chikamatsu K, Yamada H, Takaki A, Mori T, Mitarai S (2016) Evaluation of QuantiFERON-TB Gold Plus for detection of Mycobacterium tuberculosis infection in Japan. Sci Rep 6:30617. 10.1038/srep30617 PubMed PMC

Zhou G, Luo Q, Luo S, Chen H, Cai S, Guo X, He J, Xia Y, Li H, Zhou Y, Zhang Y, Song C (2023) Indeterminate results of interferon gamma release assays in the screening of latent tuberculosis infection: a systematic review and meta-analysis. Front Immunol 14:1170579. 10.3389/fimmu.2023.1170579 PubMed PMC