Diagnostic performance in active TB of QFT-Plus assay and co-expression of CD25/CD134 in response to new antigens of Mycobacterium tuberculosis

  • Ilaria SauzulloEmail author
  • Fabio Mengoni
  • Claudia Mascia
  • Paolo Pavone
  • Giulia Savelloni
  • Anna Paola Massetti
  • Miriam Lichtner
  • Vincenzo Vullo
  • Claudio M. Mastroianni
Original Investigation


The new QuantiFERON-TB Gold Plus employs modified peptides optimized to elicit an IFNγ response from CD8+ cytotoxic T lymphocytes in addition to CD4+ T cells. With a view to improve the difficult identification of TB cases, we assessed the combination of two specific immunological markers comprising IFNγ secretion and T cells co-expression of CD25 and CD134 in response to Mycobacterium tuberculosis-specific antigens. A total of 34 subjects with suspected TB and 10 age-matched HD were prospectively enrolled. Assessing the performance of QFT-Plus in terms of the TB1 and TB2 results, we found that in TB patients, the quantitative IFNγ value in TB2 was similar to that in TB1, and we did not find any differences irrespective of the disease (pulmonary or extra-pulmonary). The flow cytometric CD25/CD134 assay, allowed a more accurate differentiation between M. tuberculosis-infected and uninfected patients, with a better combination of sensitivity and specificity, especially by evaluation of CD4+ T-cell subset. All individuals with negative QFT-Plus results displayed a positive CD25/CD134 response. Overall, a positive correlation was found between T cells co-expressing CD25/CD134 and IFNγ levels in response to both QFT-Plus TB antigen tubes, as well as between the QFT-Plus TB1 and TB2 tubes. We demonstrated that both TB1 and TB2 induce a higher expression of CD25+CD134+ markers on CD4+ T cells among infected TB subjects, compared to the lower degree of CD8+ T cells, mainly induced to TB2 stimulation. We suggest that a combined use of classic QFT-Plus and specific CD25/CD134 response may be a useful means in the diagnostic workup for active TB.


QFT-Plus Tuberculosis Flow cytometry CD25/CD134 



The authors gratefully acknowledge Qiagen for providing the QFT-Plus kits, and they acknowledge Parni Nijhawan for editing the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Ilaria Sauzullo
    • 1
    Email author return OK on get
  • Fabio Mengoni
    • 1
  • Claudia Mascia
    • 1
  • Paolo Pavone
    • 1
  • Giulia Savelloni
    • 1
  • Anna Paola Massetti
    • 1
  • Miriam Lichtner
    • 1
    • 2
  • Vincenzo Vullo
    • 1
  • Claudio M. Mastroianni
    • 1
  1. 1.Department of Public Health and Infectious Diseases‘Sapienza’ UniversityRomeItaly
  2. 2.Infectious Diseases UnitSapienza UniversityLatinaItaly

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