EFFICIENCY OF USING PCR-RFLP ANALYSIS AND REAL-TIME PCR FOR DIAGNOSTICS OF DEVELOPMENTAL DUPLICATIONS, ARTHROGRIPOSIS MULTIPLEX SYNDROMES CARRIERS IN CATTLE

Abstract

The authors of the article optimized the molecular genetic methods for detecting carriers of Developmental Duplications, Arthrogriposis multiplex syndromes, determined the prevalence of the specified anomalies in the Angus, Hereford, Kazakh White-Headed, Auliekol, Kalmyk breeds (n = 37). Based on a detailed analysis of the NHLRC2 gene sequences, the PCR-RFLP analysis method was used to diagnose carriers of Developmental Duplications, the point mutation g.34618072 T> C in the 5th exon part of the NHLRC2 gene was identified using Mwol endonuclease. In order to improve the method for diagnosing the genetic anomaly Developmental Duplications, the Real Time PCR diagnostics method was developed, which allows for the determination of heterozygous carriers of the mutation g.34618072 T> C in the 5th exon part of the NHLRC2 gene within 2 hours. It is known that the syndrome - Arthrogriposis multiplex appeared as a result of an extensive deletion that covered the coding regions of three genes, ISG15, HES4, AGRN with a length of 38,000 bp. The sequences of one common forward primer and two reverse primers identifying mutant and wild types of alleles of the genes ISG15, HES4, AGRN were selected. According to the results of genotyping, the frequency of occurrence of the genetic anomaly Developmental duplication in the Angus breed was 12,5%, the syndrome Arthrogriposis multiplex in the Hereford breed was 11,1%.