ISSN Print 2500–1094
ISSN Online 2542–1204
Bulletin of RSMU
BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)
ORIGINAL RESEARCH
Detection of CFTR mutations in children with cystic fibrosis
About authors
1 DNA-Technology LLC, Moscow, Russia
2 Russian Children's Clinical Hospital, Pirogov Russian National Medical Research University, Moscow
3 Laboratory of Molecular Genetic Methods,Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Moscow
Correspondence should be addressed: Alena I. Nikiforova
Kashirskoe shosse 24, Moscow, 115478; ur.ygolonhcet-and@avorofikin
About paper
Conflict of interests: the study was conducted in collaboration with DNA-Technology staff.
Received: 2018-07-10
Accepted: 2018-08-03
Published online: 2018-08-23
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Fig. 1. А. Melting curves for different genotypes recorded during F508del (rs113993960) detection and an example of a combination of F508del and I506T (rs397508224) in the genotype. Fluorescence from FAM/НEX channels indicates the melting of probes complementary to a non-mutant or mutant gene region, respectively. The melting dynamics is recorded in the range from 25 °С to 75 °С and varies for different genotypes. 1 — mutation is absent; 2 — homozygous mutation; 3 — heterozygous mutation; 4 — a combination of F508del and I506T (the peak of the melting curves deviates from the norm) B. The sequencing chromatogram of a DNA fragment with a combination of F508del and I506T
Table 1. Primer sequences for the amplification of regions including the boundaries of CFTRdele 2,3 (21 kb)
Table 2. Results of PCR genotyping in 191 patients with cystic fibrosis
Table 3. Results of next generation sequencing of the CFTR gene in 47 patients
Note: * — represents 4 previously undescribed CFTR mutations shown in bold; ** — represents p.Ile1214Phefs (rs397508630) detected by Sanger sequencing; ? — means that candidate variants have not been identified.
Table 4. Description of 4 newly discovered variants of CFTR and patients’ phenotypes
