Discrimination of dog MHC (DLA-DRB1) alleles by high-resolution melt analysis

The function of molecules of major histocompatibility complex (MHC) is to bind antigenic peptides and display them on the cell surface for recognition by T cells. MHC polymorphism is related to suseptibility or resistance to immune-mediated and infectious diseases. High Resolution Melt (HRM) analysis is a novel and powerful technique in molecular biology for the detection of genetic polymorphisms and can be used for discrimination of MHC alleles and genotyping.
Discrimination of dog MHC alleles (DLA-DRB1) by high-resolution melt analysis.
Forty blood samples were collected from dogs referred to Small Animal Teaching Hospital of faculty of veterinary medicine, University of Tehran. After DNA extraction and amplification the exon 2 of DLA-DRB1, HRM and Melt Curve analysis was carried out. Direct sequencing was used for checking the HRM patterns.
Following the HRM and Melt curve analysis, forty samples could be classified into 8 genotypes (A to H). Most frequent was A (25.00%) and then C and E (each one 15.00%). In total, 82.5% and 17.5% of samples were heterozygote and homozygote, respectively.
In this study, the HRM technique was used for DLA-DRB1 gene alleles typing for the first time. Confirmation HRM results through sequencing results revealed that HRM can be used for discrimination of DLA-DRB1 alleles.