TSitologiya i Genetika 2023, vol. 57, no. 1, 17-25
Cytology and Genetics 2023, vol. 57, no. 1, 12–18, doi: https://www.doi.org/https://doi.org/10.3103/S009545272301005X

TRECS and KRECS measurement for screening of inborn errors of immunity

Makukh H., Boyarchuk O., Kravets V., Yarema N., Shymanska I., Kinash M., Tyrkus M., Shulhai O.

  1. Scientific medical genetic center LeoGENE, LTD, Lviv, Maksymovycha 7 g, 79031, Ukraine
  2. I. Horbachevsky Ternopil National Medical University, Ternopil, maydan Voli, 1, 46001, Ukraine
  3. Institute of Hereditary Pathology of the Ukrainian National Academy of Medical Sciences, Lviv, Lysenka, 31 a, 79008, Ukraine

SUMMARY. T-cell receptor excision rings (TRECs) and kappa-deleting excision rings (KRECs) are circular DNA molecules that are formed during the maturation of human T- and B-cells and may serve as markers of immune response cell development. To determine the TREC and KREC quantities in different biological material and their informative value for the screening of patients with inborn errors of immunity. The analysis of TREC and KREC molecules was performed by real-time polymerase chain reaction followed by the analysis of melting curves in neonatal dry spots and DNA bank of patients with inborn errors of immunity. The proposed method showed that determining the TREC copies shows 100 % sensitivity for the detection of Nijmegen syndrome and 84 % for ataxia-telangiectasia. The determination of KREC showed considerably lower sensitivity, 75 and 37 %, respectively. The calculations performed with 1,500 tests indicate the specificity of 97.6 % for the first blood spot and 97.3 % after re-collection. It would be expedient to use the method on a cohort of over 10,000 samples to clarify the specificity and to set the TREC and KREC threshold levels in our population. Determining the number of TRECs and KRECs copies is a quick and affordable method for implementation in screening programs on condition of the replication of the obtained results using a larger cohort. The determination of the TREC quantity by the proposed method showed the sensitivity of 93.6 and the 97.6 % specificity for the detection of inborn errors of immunity accompanied by T-lymphopenia.

Keywords: Real-time PCR (RT-PCR), melting curves analysis, T-cell receptor excision rings (TREC), kappa-deleting excision rings (KREC), inborn errors of immunity

TSitologiya i Genetika
2023, vol. 57, no. 1, 17-25

Current Issue
Cytology and Genetics
2023, vol. 57, no. 1, 12–18,
doi: https://doi.org/10.3103/S009545272301005X

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