Hartley Claudia, Barnett Keith C, Pettitt Louise, Forman Oliver P, Blott Sarah, Mellersh Cathryn S
Unit of Comparative Ophthalmology, Centre for Small Animal Studies, Animal Health Trust, Lanwades Park, Kentford, Newmarket CB8 7UU, Suffolk, UK.
Vet Ophthalmol. 2012 Sep;15(5):327-32. doi: 10.1111/j.1463-5224.2012.00987.x. Epub 2012 Feb 16.
To identify causative mutation(s) for congenital keratoconjunctivitis sicca and ichthyosiform dermatosis (CKCSID) in Cavalier King Charles spaniel (CKCS) dogs using a candidate gene approach.
DNA samples from 21 cases/parents were collected. Canine candidate genes (CCGs) for similar inherited human diseases were chosen. Twenty-eight candidate genes were identified by searching the Pubmed OMIM database (http://www.ncbi.nlm.nih.gov/omim). Canine orthologues of human candidate genes were identified using the Ensembl orthologue prediction facility (http://www.ensembl.org/index.html). Two microsatellites flanking each candidate gene were selected, and primers to amplify each microsatellite were designed using the Whitehead Institute primer design website (http://frodo.wi.mit.edu/primer3/). The microsatellites associated with all 28 CCGs were genotyped on a panel of 21 DNA samples from CKCS dogs (13 affected and eight carriers). Genotyping data was analyzed to identify markers homozygous in affected dogs and heterozygous in carriers (homozygosity mapping).
None of the microsatellites associated with 25 of the CCGs displayed an association with CKCSID in the 21 DNA samples tested. Three CCGs associated microsatellites were monomorphic across all samples tested.
Twenty-five CCGs were excluded as cause of CKCSID. Three CCGs could not be excluded from involvement in the inheritance of CKCSID.
采用候选基因方法鉴定骑士查理王小猎犬（CKCS）先天性干燥性角结膜炎和鱼鳞病样皮肤病（CKCSID）的致病突变。
收集21例病例/亲本的DNA样本。选择与人类相似遗传性疾病相关的犬类候选基因（CCG）。通过搜索Pubmed OMIM数据库（http://www.ncbi.nlm.nih.gov/omim）鉴定出28个候选基因。使用Ensembl直系同源物预测工具（http://www.ensembl.org/index.html）鉴定人类候选基因的犬类直系同源物。选择每个候选基因两侧的两个微卫星，并使用怀特黑德研究所引物设计网站（http://frodo.wi.mit.edu/primer3/）设计扩增每个微卫星的引物。在一组来自CKCS犬的21个DNA样本（13个患病和8个携带者）上对与所有28个CCG相关的微卫星进行基因分型。分析基因分型数据以鉴定患病犬中纯合而携带者中杂合的标记（纯合性定位）。
在测试的21个DNA样本中，与25个CCG相关的微卫星均未显示与CKCSID相关。与三个CCG相关的微卫星在所有测试样本中均为单态性。
排除25个CCG作为CKCSID的病因。不能排除三个CCG参与CKCSID的遗传。
