METHOD FOR DNA-DIAGNOSTICS OF MAJOR MUTATION DF508 IN GENE OF TRANSMEMBRANE REGULATORY PROTEIN OF MUCOVISCIDOSIS (CFTR)

摘要

A method for DNA-diagnostics of major ΔF508 mutation in gene of transmembrane regulatory protein of mucoviscidosis (CFTR) includes sampling, isolation of DNA by phenolic-chloroform method, amplification of sequence of gene CFTR containing ΔF508mutation, and subsequent analysis of amplification products by real-time polymerase chain reaction. Detection of mutation is carried out using hybridization probes of TaqMan type for normal and mutant alleles, wherein for detection of normal allele the probe HEX-AACACCAAAGATGATATT-BHQ1 is used, and for detection of allele with deletion the probe ΔF508 - Cy5-GGAAACACCAATGATATTBHQ2 is used, primers of both probes: direct - AGTTTTCCTGGATTATGCCT, reverse - AGTTGGCATGCTTTGATGAC, and according to the results of investigation the major mutation or its absence is determined.