Discrimination of dog MHC (DLA-DRB1) alleles by high-resolution melt analysis
2018
Vahedi, Seyed Milad | Jamshidi, Shahram | Lankari Mohaje, Leila | Nikbakhat Brujeni, Gholamraza
BACKGROUND: 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. OBJECTIVES: Discrimination of dog MHC alleles (DLA-DRB1) by high-resolution melt analysis. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
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