Keller Rachel L, Kania Stephen A, Hendrix Diane V H, Ward Daniel A, Abrams Kenneth
Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA.
Vet Ophthalmol. 2006 May-Jun;9(3):195-200. doi: 10.1111/j.1463-5224.2006.00466.x.
To determine the presence of serum antiretinal antibodies in sudden acquired retinal degeneration syndrome (SARDS) affected dogs and the size of the antigen to which these antibodies bind via the use of enzyme-linked immunosorbent assay (ELISA) and Western blot immunoassays.
Serum was collected from 13 dogs affected by SARDS and five dogs with normal ocular examinations.
All serum samples were subjected to ELISA with saline-soluble canine retinal tissue and Western blot analyses with SDS solubilized normal canine retinal tissue as the antigen. Antirecoverin (23 kDa) and antiheat shock cognate (65 kDa) antibodies were used as positive controls for both procedures. Affinity-purified goat antidog IgG and IgM labeled with horseradish peroxidase were used for all clinical samples and goat antirabbit IgG was used as the secondary antibody for the positive controls.
ELISA demonstrated antibody reaction with all samples. Western blot immunoassays identified multiple bands in all canine serum samples, as well as in negative controls. Approximate sizes of the bands were 25 and 50 kDa, corresponding to IgG light and heavy chains, respectively.
No antiretinal autoantibodies were identified in the serum of dogs affected by SARDS as compared to normal canine patients.
通过酶联免疫吸附测定（ELISA）和蛋白质免疫印迹免疫测定，确定患有突然获得性视网膜变性综合征（SARDS）的犬血清中抗视网膜抗体的存在以及这些抗体所结合抗原的大小。
从13只患有SARDS的犬和5只眼部检查正常的犬采集血清。
所有血清样本均用盐溶性犬视网膜组织进行ELISA检测，并用十二烷基硫酸钠（SDS）溶解的正常犬视网膜组织作为抗原进行蛋白质免疫印迹分析。抗恢复蛋白（23 kDa）和抗热休克同源蛋白（65 kDa）抗体用作这两种检测方法的阳性对照。所有临床样本均使用用辣根过氧化物酶标记的亲和纯化山羊抗犬IgG和IgM，山羊抗兔IgG用作阳性对照的二抗。
ELISA显示所有样本均有抗体反应。蛋白质免疫印迹免疫测定在所有犬血清样本以及阴性对照中均鉴定出多条条带。条带的大致大小分别为25 kDa和50 kDa，分别对应于IgG轻链和重链。
与正常犬相比，在患有SARDS的犬血清中未鉴定出抗视网膜自身抗体。
