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Updated: May 27, 2026
Simultaneous Quantification of T-Cell Receptor Excision Circles (TRECs) and K-Deleting Recombination Excision Circles (KRECs) by Real-time PCR
Published on: December 6, 2014
Neonatal screening for severe primary immunodeficiency diseases using high-throughput triplex real-time PCR
Stephan Borte1, Ulrika von Döbeln, Anders Fasth
1Division of Clinical Immunology and Transfusion Medicine, Department of Laboratory Medicine, Karolinska Institutet at Karolinska University Hospital Huddinge, Stockholm, Sweden. stephan.borte@ki.se
A new PCR method accurately quantifies T-cell receptor (TRECs) and B-cell (KRECs) markers on newborn screening cards. This aids early detection of severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia (XLA), preventing life-threatening infections.
Area of Science:
- Immunology
- Genetics
- Neonatal Screening
Background:
- Severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia (XLA) are life-threatening primary immunodeficiencies.
- Early diagnosis and treatment are crucial for preventing severe infections in infants.
- Current diagnostic criteria involve assessing T and B lymphocyte function.
Purpose of the Study:
- To develop and validate a robust triplex PCR method for quantifying TRECs and KRECs.
- To assess the utility of this method for neonatal screening of SCID and XLA.
- To identify newborns with severe T or B cell deficiencies.
Main Methods:
- A triplex PCR assay was developed to measure TREC and KREC copy numbers.
- The method was validated using 2560 anonymized newborn screening cards (Guthrie cards).
- The assay was further tested on 49 stored Guthrie cards from patients with various immunodeficiency syndromes.
Main Results:
- The PCR method successfully quantified TRECs and KRECs from a single Guthrie card punch.
- Simultaneous measurement of TREC and KREC copy numbers effectively identified patients with SCID, XLA, ataxia-telangiectasia, and Nijmegen-breakage-syndrome.
- The assay demonstrated high accuracy in detecting T and B cell deficiencies.
Conclusions:
- A validated triplex PCR method enables simultaneous measurement of TRECs and KRECs.
- This assay is effective for newborn screening of severe immunodeficiencies characterized by absent T or B cells.
- The method facilitates early diagnosis and timely intervention for conditions like SCID and XLA.
