Identification of the Mutation in DCLRE1C Gene by PCR-RFLP

Aim: Mutations in the DCLRE1C gene result in functional impairment of the Artemis protein and T/B cell development is adversely affected. As a result of this mutation, a clinic of severe combined and combined immunodeficiency (CID) generally occurs. In our region where consanguineous marriage is common, CID cases due to this mutation are frequently encountered. Therefore, suspected patients should be evaluated promptly for the relevant gene mutation. It is clear that more complicated and costly methods are used in the detection of mutations and there is a need for cheaper and faster methods. Therefore, in this study, it was aimed to determine the mutations of DCLRE1C gene exon 3 (c.194C>T; p.T65I) and exon 14 (c.1669_1670insA; p.T577Nfs*21) by using Polymerase Chain Reaction-Restriction Fragment Length
Polymorphism (PCR-RFLP) method.
Patients and Methods: The study was carried out between 2017 and 2020 and study included 14 patients followed up with DCLRE1C mutation in our clinic, 2 parents and 10 healthy controls. PCR-RFLP analysis was performed with primers containing mutation sites and approp riate restriction enzymes.
Results: As a result of the analysis, 12 patients were homozygous mutant for DCLRE1C gene exon 3, 2 parents were heterozygous for exon 3, and 2 patients were heterozygous for exon 3 and exon 14 and were found to be compound heterozygous genotype. Mutations were confirmed by Sanger DNA sequencing. Mutations in the relevant region were determined quickly and re liably by the PCR-RFLP method.
Conclusion: The study showed that the PCR-RFLP method is a cheap, safe and fast method that can be used in cases such as family screening, especially for the detection of known mutations in primary immunodeficiencies.