METHOD OF PREIMPLANTATION GENETIC TESTING FOR SPONDYLOEPIPHYSEAL DYSPLASIA Russian patent published in 2023 - IPC C12Q1/68 C12N15/00 

FIELD: biotechnology.

SUBSTANCE: method of preimplantation genetic testing of spondyloepiphyseal dysplasia which involves identifying the inheritance of the pathogenic variant c.1799G>T in the COL2A1 gene, including a dual detection system — direct and indirect, including external and internal primers presented in the following SEQ ID 1-45 is described. Direct detection is carried out using the following primers: 5'-GAGCCCCACTCATCACTGTC-3' (Fout), 5'-TGTTGGGTGGATGGATGTGG-3' (Rout), 5'-GCCAGGAAACCCATGTCA-3' (fin) paired with Fout, 5'-GGGTGGATGGATGTGGTCAG-3' (rin). Indirect detection is carried out using primers for analyzing the inheritance of molecular genetic polymorphic markers of the STR type, linked to a pathogenic variant, selected from SEQ ID 1-45, using primers aimed at those STRs whose alleles are different on all chromosomes of the parents, where external primers are designated as Fout (forward primer) and Rout (reverse primer), and the internal primers as Fin (forward primer) and Rin (reverse primer). Diagnostics is carried out in two stages of semi-nested PCR: at the first stage, multiplex PCR is carried out with external primers for STR and the COL2A1 gene, at the second stage, individual PCR of each fragment is carried out with internal primers, to analyze the inheritance of molecular genetic polymorphic markers of the STR type linked to the pathogenic variant, as well as PCR-RFLP method to determine the pathogenic variant.

EFFECT: invention expands the arsenal of agents for preimplantation genetic testing of spondyloepiphyseal dysplasia.

1 cl, 1 ex