Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, Qingdao, China.
J Appl Microbiol. 2012 Oct;113(4):737-44. doi: 10.1111/j.1365-2672.2012.05389.x. Epub 2012 Aug 2.
To develop a gold immunochromatographic test strip for on-site rapid detection of lymphocystis disease virus (LCDV).
Monodispersional colloidal gold and gold-labelled anti-LCDV monoclonal antibody (McAb) 2D11 were prepared and characterized by UV-visible spectroscopy and transmission electron microscopy. Gold-labelled probe was used as the detection antibody, and goat anti-mouse IgG at the control line and anti-LCDV McAb 1A8 at the test line of the test strip served as the capture antibody. The positive results could be easily judged by the presence of a red test line with naked eye within 10 min. The test strip, in good agreement with enzyme-linked immunosorbent assay and dot-blotting in sensitivity and LCDV detection, gave a detection limit of 1 μg ml(-1) of LCDV and was stable for 6 months at room temperature and 12 months at 4°C.
The test strip was specific, simple and convenient for rapid detection of LCDV presenting good stability and reproducibility.
This ready-to-use test strip allows on-site rapid detection of LCDV in fish without the requirement of specialized equipments and professional personnel, which could augment the practical application for diagnosis of LCDV even in disadvantage areas.
开发一种用于现场快速检测淋巴囊肿病病毒 (LCDV) 的金免疫层析测试条。
通过紫外-可见光谱和透射电子显微镜制备和表征单分散胶体金和金标记抗 LCDV 单克隆抗体 (McAb) 2D11。金标记探针用作检测抗体，测试条的检测线上的兔抗鼠 IgG 和对照线上的抗 LCDV McAb 1A8 作为捕获抗体。在 10 分钟内，通过肉眼观察到存在红色测试线，即可轻松判断出阳性结果。该测试条在灵敏度和 LCDV 检测方面与酶联免疫吸附测定和斑点印迹法高度一致，其检测限为 1μg/ml 的 LCDV，在室温下稳定 6 个月，在 4°C 下稳定 12 个月。
该测试条具有特异性、简单方便，可快速检测 LCDV，具有良好的稳定性和重现性。
这种即用型测试条可在不使用专业设备和专业人员的情况下在现场快速检测鱼类中的 LCDV，即使在条件不利的地区，也可增强 LCDV 诊断的实际应用。
