Zhang Liping, Li Dengfeng, Liu Lianguo, Fang Jing, Xu Ran, Zhang Ge
School of Marine Sciences, Ningbo University, Fenghua Road, Ningbo, Zhejiang 315211, PR China.
School of Marine Sciences, Ningbo University, Fenghua Road, Ningbo, Zhejiang 315211, PR China.
J Virol Methods. 2015 Sep 1;221:39-45. doi: 10.1016/j.jviromet.2015.04.016. Epub 2015 Apr 22.
A colloidal gold immunochromatographic strip (ICS) test based on a competitive format was developed for the rapid detection of soft-shelled turtle systemic septicemia spherical virus (STSSSV) in turtle and fecal samples. Specific egg yolk antibodies (IgY) against STSSSV were labeled with colloidal gold and used as probes in the one-step test strip. Antigen (STSSSV) and goat anti-chicken IgY were drawn on the nitrocellulose membrane as the test line and control line, respectively. When STSSSV standard samples (0-100μg/mL) were detected by the strips, the visual limit of detection (LOD) was found to be 50.0μg/mL. The ICS test showed high stability; the strips were stable for at least 3 months at 4°C without significant loss of activity. There was no obvious cross-reactivity with other aquatic pathogens. The assay can be performed within 5-10min. Analysis of STSSSV in turtle samples revealed that data obtained from the ICS test were in a good agreement with those obtained by ELISA. The positive results of fecal samples suggested that this method could be used to detect STSSSV while protecting the animals' welfare. The ICS assay does not need specialized equipment or a technician and can be used as a reliable, rapid, cost-effective and convenient qualitative tool for on-site diagnosis.
开发了一种基于竞争模式的胶体金免疫层析试纸条（ICS）检测方法，用于快速检测龟类及粪便样本中的中华鳖全身性败血病球形病毒（STSSSV）。针对STSSSV的特异性蛋黄抗体（IgY）用胶体金标记，并用作一步式试纸条中的探针。抗原（STSSSV）和山羊抗鸡IgY分别绘制在硝酸纤维素膜上作为检测线和对照线。当用试纸条检测STSSSV标准样品（0-100μg/mL）时，目视检测限（LOD）为50.0μg/mL。ICS检测显示出高稳定性；试纸条在4°C下至少稳定3个月，活性无明显损失。与其他水生病原体无明显交叉反应。该检测可在5-10分钟内完成。对龟类样本中STSSSV的分析表明，ICS检测获得的数据与ELISA获得的数据高度一致。粪便样本的阳性结果表明，该方法可用于检测STSSSV，同时保护动物福利。ICS检测不需要专门设备或技术人员，可作为一种可靠、快速、经济高效且方便的现场诊断定性工具。
