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

DIAZ S.; ECHEVERRIA M.G.; IT V.; POSIK D.M.; ROGBERG MUÑOZ A.; PENA N.; PERAL GARCIA P.; VEGA PLA J.L.; GIOVAMBATTISTAG.

TISSUE ANTIGENS

International Bookksellers Publishers

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 PCR-SSCP. 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.