IDENTIFICATION OF HOLSTEIN COWS CARRIERS OF COMPLEX VERTEBRAL MALFORMATION BY HIGH RESOLUTION MELTING CURVES (HRM)

• Main Author: Branda Sica, Andrea
• Publication Date: 2019
• Other Authors: Nicolini, Paula, Federici, María Teresa, Llambí, Silvia
• Format: Article
• Language: eng
• Source: Archives of Veterinary Science (Online)
• Download full: https://revistas.ufpr.br/veterinary/article/view/65494
• topic: Holstein; CVM; high-resolution dissociation curve.

The objective of this study was the optimization and implementation of a reliable and economical molecular screening method for the detection of the mutant allele of CVM (complex vertebral malformation, c.559G>T, SLC35A3) by HRM analysis, as well as analyzing its existence in a representative sample of Holstein cows from the Milk Genomic DNA Bank of Uruguay. The optimization of the HRM methodology in the RotorGene™ 6000 equipment (Corbett Life Science, Australia) by amplification of the 79 bp PCR products clearly differentiated the two genotypes: homozygous, wild type: GG; and heterozygous, carrier for the mutation CVM: GT (c.559G>T; SLC35A3). In the analyzed sample, the frequency of the mutant allele (T) for CVM was high (q = 0.032), with a prevalence of carrier cows of 6.45%. It is concluded that the PCR-HRM analysis is a fast, easily interpretable, low cost, and highly accurate technique for the detection of this mutation in Holstein cattle, which may be implemented in genetic selection programs.