Zhang LiPing, Li DengFeng, Liu LianGuo, Zhang Ge
School of Marine Sciences, Ningbo University, Fenghua Road, Ningbo, Zhejiang 315211, PR China.
Dis Aquat Organ. 2015 Nov 17;117(1):21-9. doi: 10.3354/dao02921.
Swimming crab reovirus (SCRV) is the causative agent of a serious disease with high mortality in cultured Portunus trituberculatus. A rapid immunochromatographic assay (ICA) was developed in a competitive assay format and optimized for the detection of SCRV. The gold probe-based ICA test comprised SCRV antigen and goat anti-chicken egg yolk antibody (IgY) sprayed onto a nitrocellulose membrane as the test line and control line, respectively. IgY-gold complexes were deposited onto the conjugate pad as detector reagents. The method showed high specificity with no cross-reactivity with other related aquatic pathogens. The detection limit of the ICA strip was 50 µg ml⁻¹. To evaluate the performance of the ICA test, the strip and an enzyme-linked immunosorbent assay (ELISA) were applied to the same samples (n = 90 crabs). The strip successfully detected SCRV in all of the artificially infected samples. Furthermore, the ICA strip and ELISA tests had high consistency (98.28%). The strip assay requires no instruments and has a detection time of less than 10 min. It is portable and easy to perform in the field. These results indicated that the developed strip could be a promising on-site tool for screening pooled crabs to confirm SCRV infection or disease outbreaks.
三疣梭子蟹呼肠孤病毒（SCRV）是养殖三疣梭子蟹中一种导致高死亡率严重疾病的病原体。开发了一种竞争检测形式的快速免疫层析检测法（ICA），并针对SCRV的检测进行了优化。基于金探针的ICA检测包括分别喷涂在硝酸纤维素膜上作为检测线和对照线的SCRV抗原和山羊抗鸡卵黄抗体（IgY）。IgY-金复合物作为检测试剂沉积在结合垫上。该方法显示出高特异性，与其他相关水生病原体无交叉反应。ICA试纸条的检测限为50 µg ml⁻¹。为了评估ICA检测的性能，将试纸条和酶联免疫吸附测定法（ELISA）应用于相同的样本（n = 90只螃蟹）。试纸条成功检测出所有人工感染样本中的SCRV。此外，ICA试纸条和ELISA检测具有高度一致性（98.28%）。试纸条检测无需仪器，检测时间不到10分钟。它便于携带，易于在现场进行操作。这些结果表明，所开发的试纸条可能是一种很有前景的现场工具，用于筛选大批螃蟹以确认SCRV感染或疾病爆发。
