PREIMPLANTATION GENETIC TESTING METHOD FOR ACHONDROPLASIA Russian patent published in 2023 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: method for diagnosing achondroplasia monogenic disease under conditions of preimplantation genetic testing (PGT) is described. The method is carried out using a test system for diagnosing the pathogenic variant NC_000004.11:g.1806119G>A (NM_000142.4:c.1138G>A, p.Gly380Arg) in FGFR3 gene. The method includes a double detection system — direct and indirect, where direct detection is carried out using primers for amplification: external: direct: 5'-CTGGCGTTACTGACTGCG-3', reverse: 5'-GGGAGATCTTGTGCACGG-3', internal for wild-type allele detection: direct: 5'-GTATGCAGGCATCCTCAGCGAC-3' paired with external reverse, internal for the detection of the mutant allele: direct: 5'-CATGTCTTTGCAGCCGAG-3' paired with an external reverse. Indirect detection is carried out using primers for the analysis of the inheritance of molecular genetic markers of the STR type linked to the pathogenic variant, selected from SEQ ID NO 1-65, while using primers directed to those STRs whose alleles are different on the chromosomes of at least one of the parents, where the outer primers are designated as Fout (forward primer) and Rout (reverse primer), and the internal primers are designated as Fin (forward primer) and Rin (reverse primer), while the diagnosis is carried out in two stages of semi-nested PCR: at the first stage, multiplex PCR with external primers, at the second stage, individual PCR of each fragment is carried out with internal primers for STR, as well as the PCR-RFLP method to determine the pathogenic variant in FGFR3 gene.

EFFECT: invention allows to carry out specific and efficient preimplantation genetic testing of achondroplasia for families with a high risk of this disease developing in future children.

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