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

FIELD: biotechnology.

SUBSTANCE: method for diagnosing the monogenic disease familial glomerular hematuria (Alport syndrome) under conditions of preimplantation genetic testing is described. The inheritance of the pathogenic variant NC_000023.10:g.107850123G>A (NM_033380.3:c.2395+1G>A) in the COL4A5 gene is detected by double detection — direct and indirect. Direct detection is carried out using amplification primers:

external primers:

5'-CTCAAGGGTGAACCAGGATT-3' (Fout),

5'-ACAGAATGGCAGCATAGGGT-3' (Rout),

internal primers:

5'-GCTCCCTGGACCAAACG-3' (fin_mm_wt),

5'-GGCTCCCTGGACCAGAG-3' (fin_mm_mut).

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 1-27. Primers directed to those STRs whose alleles are different on the chromosomes of the parent-carrier of the mutation are used. Diagnosis is carried out in two stages of semi-nested PCR: at the first stage, multiplex PCR is performed with external primers, at the second stage, individual PCR of each fragment is performed with internal primers for STR. The PCR-RFLP method is also used to determine the pathogenic variant in the COL4A5 gene. The method has high specificity and efficiency, as well as versatility in relation to biosamples of various types: single cells, the product of whole genome amplification, total DNA isolated from different tissues.

EFFECT: method can be used not only for the detection of a specific pathogenic variant, but also for any other pathogenic variant or several in the COL4A5 gene using indirect detection.

1 cl, 2 tbl, 1 ex