Abstract
The linkage phase of marker C04107 was evaluated before implementation of the marker in a diagnostic test. Blood samples from 68 dogs were collected and genotyped by PCR. Two alleles were detected with sizes of 160 bp and 164 bp and allele frequencies of 0.45 and 0.55 respectively. Genotyping revealed that 35 dogs were heterozygous (51.5%), 22 dogs were homozygous for the normal allele (32.3%) and 11 dogs were homozygous for the disease allele (16.2%). Liver biopsies were taken from 14 selected dogs and the copper content was evaluated histologically. Biopsies from 8 dogs homozygous for the disease allele showed many copper granules along with single cell necrosis, haemosiderosis and cellular infiltration. In liver biopsies from 6 dogs genotyped to be heterozygous or homozygous for the normal allele, copper granules were absent or moderate in number and no lesions were present. The survey demonstrates that the linkage phase of marker C04107 in the Danish population of Bedlington terriers is similar to the linkage phase detected in other countries. Thus, the marker can be used in a diagnostic test for copper toxicosis in Denmark.
Keywords: DNA-marker, liver disease
Sammendrag
Koblingsfasen af markør C04107 blev evalueret forud for implementering af markøren i en diagnostisk test i Danmark. Blodprøver fra 68 hunde blev indsamlet og genotypet ved hjælp af PCR. Ved genotypningen fremkom to alleler med størrelser på henholdsvis 160bp og 164bp samt allelfrekvenser på henholdsvis 0.45 og 0.55. Genotypningen viste, at 35 hunde var heterozygote (51.5%), 22 hunde var homozygote for den raske allel (32.3%), og 11 hunde var homozygote for den defekte allel (16.2%). Leverbiopsier blev udtaget fra 14 udvalgte hunde, og kobberindholdet vurderet histologisk. I biopsier fra 8 hunde, der var homozygote for den defekte allel, sås mange kobberholdige granula, nekrose af enkelte celler, hæmosiderose og infiltration med inflammationsceller. I leverbiopsier fra 6 hunde, der var heterozygote eller homozygote for den raske allel, sås et varierende antal af kobbergranula fra ingen til et moderat antal og ingen læsioner. Undersøgelsen demonstrerer, at koblingsfasen mellem markør C04107 og sygdomsgenet er den samme i den danske Bedlington terrier population som i de øvrige undersøgte populationer i USA og Europa. Markøren er derfor anvendelig i en diagnostisk test for kobertoxicose i Danmark.
Full Text
The Full Text of this article is available as a PDF (988.0 KB).
References
- Brewer GJ, Dick RD, Schall W, Yuzbasian-Gurkan V, Mullaney TP, Pace C, Lindgren J, Thomas M, Padgett G. Use of zink acetate to treat copper toxicosis in dogs. Journal of the American Veterinary Medical Association. 1992;201:564–568. [PubMed] [Google Scholar]
- Hardy RM, Stevens JB, Stowe CM. Cronic progressive hepatitis in Bedlington terriers associated with elevated liver copper concentrations. Minnesota Veterinarian. 1975;15:13–24. [Google Scholar]
- Holmes NG, Herrtage ME, Ryder EJ, Binns MM. DNA marker C04107 for copper toxicosis in a population of Bedlington terriers in the United Kingdom. Veterinary Record. 1998;142:351–352. doi: 10.1136/vr.142.14.351. [DOI] [PubMed] [Google Scholar]
- Johnson GF, Sternlieb I, Twedt DC, Grushoff PS, Scheinberg IH. Inheritance of Copper Toxicosis in Bedlington Terriers. American Journal of Veterinary Research. 1980;41:1865–1866. [PubMed] [Google Scholar]
- Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research. 1988;16:1215–1215. doi: 10.1093/nar/16.3.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rothhuizen J, Ubbink GJ, van Zon P, Teske E, van den Ingh TSGAM, Yuzbasiyan-Gurkan V. Diagnostic value of a microsatellite DNA marker for copper toxicosis in West-European Bedlington terriers and incidence of the disease. Animal Genetics. 1999;30:190–194. doi: 10.1046/j.1365-2052.1999.00451.x. [DOI] [PubMed] [Google Scholar]
- Su L, Ravanshad S, Owen CA, Jr, McCall JT, Zollman PE, Hardy RM. A comparison of copper loading disease in Bedlington terriers and Wilson’s disease in humans. American Journal of Physiology. 1982;243:G226–G230. doi: 10.1152/ajpgi.1982.243.3.G226. [DOI] [PubMed] [Google Scholar]
- Teske E, Brinkhuis BGAM, Bode P, van den Ingh Th.SGAM, Rothuizen J. Cytological detection of copper for diagnosis of inherited copper toxicosis in Bedlington terriers. Veterinary Record. 1992;131:30–32. doi: 10.1136/vr.131.2.30. [DOI] [PubMed] [Google Scholar]
- Thornburg LP, Beissenherz M, Dolan M, Raisbeck MF. Histochemical demonstration of copper and copper-associated protein in the canine liver. Veterinary Pathology. 1985;22:327–332. doi: 10.1177/030098588502200405. [DOI] [PubMed] [Google Scholar]
- Twedt DC, Sternlieb I, Gilderson SR. Clinical, morphologic, and chemical studies on copper toxicosis of Bedlington terriers. Journal of the American Veterinary Medical Association. 1979;175:269–275. [PubMed] [Google Scholar]
- Ubbink GJ, Van de Broek J, Hazewinkel HAW, Rothuizen J. Cluster analysis of the genetic heterogeneity and disease distributions in purebred dog populations. Veterinary Record. 1998;142:209–213. doi: 10.1136/vr.142.9.209. [DOI] [PubMed] [Google Scholar]
- Van de Sluis BJA, Breen M, Manoj N, Van Wolferen M, De Jong P, Binns MM, Pearson PL, Kuipers J, Rothuizen J, Cox DW, Wijmenga C, Van Oost BA. Genetic mapping of the copper toxicosis locus in Bedlington terriers to dog chromosome 10, in a region syntenic to human chromosome region 2p13–p16. Human Molecular Genetics. 1999;8:501–507. doi: 10.1093/hmg/8.3.501. [DOI] [PubMed] [Google Scholar]
- Weber JL, May PE. Abundant Class of Human DNA Polymorphisms Which Can Be Typed Using the Polymerase Chain Reaction. American Journal of Human Genetics. 1989;44:388–396. [PMC free article] [PubMed] [Google Scholar]
- Yuzbasiyan-Gurkan V, Blanton SH, Cao Y, Ferguson P, Li J, Venta PJ, Brewer GJ. Linkage of a microsatellite marker to the canine copper toxicosis locus in Bedlington Terriers. American Journal of Veterinary Research. 1997;58:1–5. [PubMed] [Google Scholar]
- Yuzbasiyan-Gurkan V, Wagnitz S, Blanton SH, Brewer GJ. Linkage Studies of the Esterase D and Retinoblastoma Genes to Copper Toxicosis: A Model for Wilson Disease. Genomics. 1993;15:86–90. doi: 10.1006/geno.1993.1013. [DOI] [PubMed] [Google Scholar]