Development of an ELA-DRA gene typing method based on pyrosequencing technology

DÍAZ, S; ECHEVERRÍA, G; IT, V; POSIK, DM; ROGBERG-MUÑOZ, A; PERAL-GARCÍA, P; VEGA-PLA, JL; GIOVAMBATTISTA, G

TISSUE ANTIGENS

Blackwell Publishing - Wiley Inter Science

Lugar: Parkville, Victoria 3052 - Australia; Año: 2008 vol. 72 p. 464 - 468

0001-2815

The polymorphism of equine lymphocyte antigen (ELA) class II DRA gene had been detected by polymerase chain reaction–single-strand conformational polymorphism (PCR-SSCP) and reference strand-mediated conformation analysis. These methodologies allowed to identify 11 ELA-DRA exon 2 sequences, three of  which are widely distributed among domestic horse breeds. Herein, we describe the development of a pyrosequencing-based method applicable to ELA-DRA typing,  by screening samples from eight different horse breeds previously typed by PCRSSCP. This sequence-based method would be useful in high-throughput genotyping of major histocompatibility complex genes in horses and other animal species, making this system interesting as a rapid screening method for animal genotyping of immune-related genes. DRA gene had  been detected by polymerase chain reaction–single-strand conformational polymorphism (PCR-SSCP) and reference strand-mediated conformation analysis. These methodologies allowed to identify 11 ELA-DRA exon 2 sequences, three of  which are widely distributed among domestic horse breeds. Herein, we describe the development of a pyrosequencing-based method applicable to ELA-DRA typing,  by screening samples from eight different horse breeds previously typed by PCRSSCP.  This sequence-based method would be useful in high-throughput genotyping of major histocompatibility complex genes in horses and other animal species, making this system interesting as a rapid screening method for animal genotyping of immune-related genes.