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开发一种胶体金免疫层析法(GICA),用于快速检测来自中国商业养殖甲壳类动物中的河蟹螺原体。

Development of a colloidal gold immunochromatographic assay (GICA) for the rapid detection of Spiroplasma eriocheiris in commercially exploited crustaceans from China.

作者信息

Liu H, Xiu Y, Xu Y, Tang M, Li S, Gu W, Meng Q, Wang W

机构信息

Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, Nanjing, Jiangsu, China.

Co-Innovation Center for Marine Bio-Industry Technology of Jiangsu Province, Lianyungang, Jiangsu, China.

出版信息

J Fish Dis. 2017 Dec;40(12):1839-1847. doi: 10.1111/jfd.12657. Epub 2017 Jun 20.

Abstract

Spiroplasma eriocheiris is an emerging pathogen in freshwater crustaceans. In recent years, Eriocheir sinensis, Procambarus clarkii, Litopenaeus vannamei, Macrobrachium rosenbergii and Macrobrachium nipponense had been infected by this pathogen in China. An immunochromatographic strip test using gold nanoparticles was developed for rapidly detecting this pathogen. The strip test based on the principle of sandwich immunoassay by the specific combination between the pathogen and polyclonal antibody on a nitrocellulose membrane. Positive samples were displayed as red lines at the test and control zones of the nitrocellulose membrane, while negative samples resulted in a red line only at the control zone. The limit of detection was proved to be 10 Color Change Unit/ml. The test strip could be visually detected within 15 min and do not have cross-reaction with other aquatic bacteria. This test strip allows on-site rapid detection of S. eriocheiris in crustacean without the requirement of specialized equipment and professional personnel. The one-step test strips developed in our study had high sensitivity, specificity, reproducibility and stability. In conclusion, this method was proved to be convenient, feasible, rapid and effective for detecting S. eriocheiris.

摘要

中华绒螯蟹螺原体是淡水甲壳类动物中的一种新兴病原体。近年来,中国的中华绒螯蟹、克氏原螯虾、凡纳滨对虾、罗氏沼虾和日本沼虾都受到了这种病原体的感染。开发了一种使用金纳米颗粒的免疫层析试纸条来快速检测这种病原体。该试纸条基于夹心免疫测定原理,通过病原体与硝酸纤维素膜上的多克隆抗体特异性结合。阳性样品在硝酸纤维素膜的测试区和对照区显示为红线,而阴性样品仅在对照区产生红线。检测限被证明为10个颜色变化单位/毫升。该试纸条可在15分钟内肉眼检测,且与其他水生细菌无交叉反应。这种试纸条无需专门设备和专业人员即可在现场快速检测甲壳类动物中的中华绒螯蟹螺原体。我们研究中开发的一步式试纸条具有高灵敏度、特异性、重现性和稳定性。总之,该方法被证明对检测中华绒螯蟹螺原体方便、可行、快速且有效。

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