Development of an ELA-DRA gene typing method based on pyrosequencing technology.
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.
Díaz
S
S
Centro de Investigaciones en Genética Básica y Aplicada, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Buenos Aires, Argentina. sdiaz@fcv.unlp.edu.ar
Echeverría
M G
MG
It
V
V
Posik
D M
DM
Rogberg-Muñoz
A
A
Pena
N L
NL
Peral-García
P
P
Vega-Pla
J L
JL
Giovambattista
G
G