Application of molecular-genetic approaches in Cftr gene variant diagnostics: historical perspective and personal experience

Aim. To analyze the evolution of molecular-genetic methods for cystic fibrosis diagnosis, assess their effectiveness, advantages, and limitations, and determine optimal approaches for detecting CFTR gene variants. Methods. An analysis of the national registry of cystic fibrosis patients was conducted, along with laboratory studies performed at the Institute of Hereditary Pathology and the Scientific Medical-Genetic Center “LeoGene”. The study investigated methods including heteroduplex analysis, restriction analysis, MLPA, INNO-LiPA CFTR, TaqMan, Sanger sequencing, and NGS. Results. It was shown that the combined use of first-line methods identifying major population variants, along with MLPA, allows the detection of up to 70 % of CFTR variants. NGS provides a more detailed mutation analysis, including rare variants. Conclusions. The most effective approach is a stepwise strategy, incorporating initial screening for common variants followed by NGS analysis in case of a negative result. The use of international HGVS standards and databases such as CFTR2 and NCBI is crucial for the unification of diagnostic conclusions.