Development and application of antibody microarray for lymphocystis disease virus detection in fish

被引:9
|
作者
Sheng, Xiuzhen [1 ]
Xu, Xiaoli [1 ]
Zhan, Wenbin [1 ]
机构
[1] Ocean Univ China, Lab Pathol & Immunol Aquat Animals, Qingdao 266003, Peoples R China
基金
中国国家自然科学基金;
关键词
Lymphocystis disease virus (LCDV); Antibody microarray; Agarose gel; Virus detection; SPOT-SYNDROME-VIRUS; FLOUNDER PARALICHTHYS-OLIVACEUS; PROTEIN MICROARRAY; FLUORESCENT;
D O I
10.1016/j.jviromet.2013.02.015
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease affecting marine and freshwater fish worldwide. Here an antibody microarray was developed and employed to detect LCDV in fish. Rabbit anti-LCDV serum was arrayed on agarose gel-modified slides as capture antibody, and Cy3-conjugated anti-LCDV monoclonal antibody (MAbs) was added as detection antibody. The signals were imaged with a laser chip scanner and analyzed by corresponding software. To improve the sensitivity, different substrate binders (poly-L-lysine, MPTS, aldehyde, APES and agarose gel modified slides, and commercially available amino-modified slides), markers (fluorescein isothiocyanate, Cy3, horseradish peroxidase, biotin or colloidal gold) conjugated to anti-LCDV Mabs, and storage time of the antibody were assessed. The results showed that the antibody microarrays based on agarose gel-modified slides gave a lower detection limit of 0.55 mu g/ml of LCDV when Cy3 and HRP conjugated anti-LCDV MAbs were used as detection antibody; and the lowest detectable LCDV protein concentration was 0.0686 mu g/ml when streptavidin-biotin conjugated to anti-LCDV MAbs served as detection antibody. The developed antibody microarray proved to have a high specificity for LCDV detection and a shelf-life of more than 8 months at -20 degrees C. Furthermore, the LCDV detection results of the microarray in fish gills or fins (n = 50) presented a concordance rate of 100% with enzyme-linked immunosorbent assay (ELISA) and 98% with immunofluorescence assay technique (IFAT). These results revealed that the developed antibody microarray could serve as an effective tool for diagnostic and epidemiological studies of LCDV in fish. (c) 2013 Elsevier B.V. All rights reserved.
引用
收藏
页码:243 / 249
页数:7
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