Detection of complex vertebral malformation carriers in Slovak Holstein cattle by high resolution melting analysis

Abstract

The objective of this study was to apply high resolution melting analysis in the detection of complex vertebral malformation (CVM) carriers in Hosltein cattle. A total of 47 animals of Holstein cattle were included in this study. Genomic DNA was extracted from blood, hair follicles and sperm by commercial QIAamp® DNA Mini kit. The amplification and high resolution melting analysis (HRMA) was done by commercial SensiMixTM HRM kit. The confirmation of sensitivity of this method was done by PCR-PIRA method and sequencing. Four samples of heterozygous genotype GT for causal mutation in the bovine solute carrier family 35 member 3 gene (SLC35A3), which is responsible for CVM disease, were detected. Our results demostrated that the use of HRMA for genotyping of mutant allele T for SLC35A3 gene in Holstein cattle is an effective method for the selection of carriers of CVM disease.

Keywords

• complex vertebral malformation (CVM)
• high resolution melting analysis (HRMA)
• Holstein cattle
• genomic DNA