SET OF OLIGONUCLEOTIDE PRIMERS AND PROBES FOR GENETIC TYPING OF POLYMORPHOUS DNA LOCI ASSOCIATED WITH A RISK OF PROGRESSION OF SPORADIC FORM OF ALZHEIMER'S DISEASE IN RUSSIAN POPULATIONS

摘要

FIELD: genetics; molecular biology; medicine.;SUBSTANCE: invention relates to a method for analysis of genetic polymorphism in DNA of ApoE, ApoJ and GAB2 and a set of oligonucleotide primers and probes. Described method includes the steps of: (a) amplification of polymorphic gene fragments of ApoE, ApoJ and GAB2 by multiplex "circular" two-step polymerase chain reaction, wherein DNA sample is used as the matrix for amplification; during I step of PCR a set of primers with sequences given in SEQ ID NO: 9, 10, 13, 14, 17, 18 is used; during II step of PCR a set of primers with sequences given in SEQ ID NO: 11, 12, 15, 16, 19, 20 is used, and deoxyuridine triphosphate, marked with Su-5 fluorescent dye, resulting in formation of fluorescent marked PCR-product; (b) immobilization of oligonucleotide probes, sequence of which are presented in SEQ ID NO: 1-8, on a solid substrate; (c) hybridization of the marked PCR-product with immobilized oligonucleotides presented in SEQ ID NO: 1-8; (d) registration and interpretation of hybridization results.;EFFECT: inventions provide the possibility of determining nucleotide composition in four polymorphous positions: ApoE (rs429358, rs7412), ApoJ (rs11136000) and GAB2 (rs2373115), that, in its turn, enables to determine predisposition to sporadic form of Alzheimer's disease in the population of Russia.;2 cl, 2 dwg, 3 tbl, 4 ex