Genomic screening for fucosidosis in English Springer Spaniels.
OBJECTIVE
To develop a robust molecular genetic test for alpha-L-fucosidosis in English Springer Spaniels and to screen dogs from the United Kingdom and United States for the mutant allele.


METHODS
35 English-bred English Springer Spaniels, 60 American-bred English Springer Spaniels, and 1 affected dog and its parents from a family of English Springer Spaniels in Colorado.


METHODS
Polymerase chain reaction analysis was used to amplify the mutated region in the gene encoding alpha-L-fucosidase. High guanine-cytosine (GC) content of the region required use of an amplification buffer with high pH. Mutant and normal alleles were separated by polyacrylamide gel electrophoresis. Molecular genetic test results were compared with enzyme data.


RESULTS
A 262-bp PCR product was amplified from normal dogs and compared with a 248-bp product from affected dogs. Carriers had 1 copy of each allele, distinguishable by the 14-bp size difference. Two carriers among the English-bred dogs were identified by use of enzyme and genomic DNA analyses. The molecular defect in dogs from Colorado was proven to be the same as that in British and Australian dogs. None of the other 60 American-bred dogs carried the mutant allele.


CONCLUSIONS
A PCR method that can be used to identify dogs affected with or carriers of the autosomal recessive disease fucosidosis was established. Amplification was achieved within a GC-rich region, using a method that may be useful in overcoming amplification problems in GC-rich areas within other genes. Using this test, fucosidosis can be controlled and ultimately eradicated from the English Springer Spaniel population.
Skelly
B J
BJ
Josephine Deubler Genetic Disease Testing Laboratory, Section of Medical Genetics, University of Pennsylvania, Philadelphia 19104-6010, USA.
Sargan
D R
DR
Winchester
B G
BG
Smith
M O
MO
Herrtage
M E
ME
Giger
U
U
D000818:Animals; D001315:Australia; D020029:Base Pairing; D003120:Colorado; D003596:Cytosine; D004283:Dog Diseases / Q000175:diagnosis / Q000235:genetics*; D004285:Dogs; D005260:Female; D005645:Fucosidosis / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*; D005784:Gene Amplification; D006580:Genetic Carrier Screening; D005820:Genetic Testing / Q000379:methods / Q000662:veterinary*; D006147:Guanine; D008297:Male; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D013045:Species Specificity; D006113:United Kingdom; D005644:alpha-L-Fucosidase / Q000097:blood / Q000235:genetics*
D006147:Guanine; D003596:Cytosine; D005644:alpha-L-Fucosidase