PMID- 25485546 OWN - NLM STAT- MEDLINE DCOM- 20160112 LR - 20181113 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 9 IP - 12 DP - 2014 TI - The TREC/KREC assay for the diagnosis and monitoring of patients with DiGeorge syndrome. PG - e114514 LID - 10.1371/journal.pone.0114514 [doi] LID - e114514 AB - DiGeorge syndrome (DGS) presents with a wide spectrum of thymic pathologies. Nationwide neonatal screening programs of lymphocyte production using T-cell recombination excision circles (TREC) have repeatedly identified patients with DGS. We tested what proportion of DGS patients could be identified at birth by combined TREC and kappa-deleting element recombination circle (KREC) screening. Furthermore, we followed TREC/KREC levels in peripheral blood (PB) to monitor postnatal changes in lymphocyte production. METHODS: TREC/KREC copies were assessed by quantitative PCR (qPCR) and were related to the albumin control gene in dry blood spots (DBSs) from control (n = 56), severe immunodeficiency syndrome (SCID, n = 10) and DGS (n = 13) newborns. PB was evaluated in DGS children (n = 32), in diagnostic samples from SCID babies (n = 5) and in 91 controls. RESULTS: All but one DGS patient had TREC levels in the normal range at birth, albeit quantitative TREC values were significantly lower in the DGS cohort. One patient had slightly reduced KREC at birth. Postnatal DGS samples revealed reduced TREC numbers in 5 of 32 (16%) patients, whereas KREC copy numbers were similar to controls. Both TREC and KREC levels showed a more pronounced decrease with age in DGS patients than in controls (p < 0.0001 for both in a linear model). DGS patients had higher percentages of NK cells at the expense of T cells (p < 0.0001). The patients with reduced TREC levels had repeated infections in infancy and developed allergy and/or autoimmunity, but they were not strikingly different from other patients. In 12 DGS patients with paired DBS and blood samples, the TREC/KREC levels were mostly stable or increased and showed similar kinetics in respective patients. CONCLUSIONS: The combined TREC/KREC approach with correction via control gene identified 1 of 13 (8%) of DiGeorge syndrome patients at birth in our cohort. The majority of patients had TREC/KREC levels in the normal range. FAU - Fronkova, Eva AU - Fronkova E AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Klocperk, Adam AU - Klocperk A AD - Department of Immunology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Svaton, Michael AU - Svaton M AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Novakova, Michaela AU - Novakova M AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Kotrova, Michaela AU - Kotrova M AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Kayserova, Jana AU - Kayserova J AD - Department of Immunology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Kalina, Tomas AU - Kalina T AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Keslova, Petra AU - Keslova P AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Votava, Felix AU - Votava F AD - Department of Pediatrics, 3rd Medical School, Charles University Prague and University Hospital Kralovske Vinohrady, Prague, Czech Republic. FAU - Vinohradska, Hana AU - Vinohradska H AD - Department of Clinical Biochemistry, Children Hospital, Faculty of Medicine, Masaryk University Brno, Brno, Czech Republic. FAU - Freiberger, Tomas AU - Freiberger T AD - Department of Clinical Immunology and Allergology, Medical Faculty, and Central European Institute of Technology, Masaryk University, Brno, Czech Republic; Molecular Genetics Lab, Centre for Cardiovascular Surgery and Transplantation, Brno, Czech Republic. FAU - Mejstrikova, Ester AU - Mejstrikova E AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Trka, Jan AU - Trka J AD - CLIP, Department of Paediatric Haematology/Oncology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. FAU - Sediva, Anna AU - Sediva A AD - Department of Immunology, 2nd Medical School, Charles University Prague and University Hospital Motol, Prague, Czech Republic. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20141208 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (DNA, Circular) SB - IM MH - Adolescent MH - Biological Assay MH - Cells, Cultured MH - Child MH - Child, Preschool MH - DNA, Circular/*genetics MH - DiGeorge Syndrome/*diagnosis/genetics/immunology MH - Female MH - Humans MH - Immunologic Deficiency Syndromes/*diagnosis/genetics/immunology MH - Infant MH - Infant, Newborn MH - Male MH - *Neonatal Screening MH - Real-Time Polymerase Chain Reaction MH - Severe Combined Immunodeficiency/*diagnosis/genetics/immunology MH - T-Lymphocytes/*immunology PMC - PMC4259354 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2014/12/09 06:00 MHDA- 2016/01/13 06:00 PMCR- 2014/12/08 CRDT- 2014/12/09 06:00 PHST- 2014/06/04 00:00 [received] PHST- 2014/11/10 00:00 [accepted] PHST- 2014/12/09 06:00 [entrez] PHST- 2014/12/09 06:00 [pubmed] PHST- 2016/01/13 06:00 [medline] PHST- 2014/12/08 00:00 [pmc-release] AID - PONE-D-14-24503 [pii] AID - 10.1371/journal.pone.0114514 [doi] PST - epublish SO - PLoS One. 2014 Dec 8;9(12):e114514. doi: 10.1371/journal.pone.0114514. eCollection 2014.