科研動態(tài)

針對魚類致病性腸道弧菌（Vibrio ichthyoenteri）、鰻弧菌（Vibrio anguillarum）、副溶血弧菌（Vibrio parahaemolyticus）、溶藻弧菌（Vibrio alginolyticus）、河流弧菌（Vibrio fluvialis）和哈維氏弧菌（Vibrio harveyi），制備了牙鲆（Paralichthys olivaceus）抗血清，提取了各弧菌的外膜蛋白、胞外產(chǎn)物、鞭毛蛋白和全菌蛋白，分析了牙鲆抗血清與各抗原蛋白的免疫反應(yīng)，發(fā)現(xiàn)各弧菌抗血清與各自的抗原反應(yīng)最強(qiáng)，與其他弧菌抗原有不同程度的交叉反應(yīng)。將6種弧菌的抗原組分固定于芯片載體形成微陣列，構(gòu)建了其血清學(xué)診斷抗原芯片，使用辣根過氧化物酶標(biāo)記的抗牙鲆IgM單抗2D8作為檢測抗體，確定了檢測結(jié)果判讀方法；將抗原芯片應(yīng)用于牙鲆和大菱鲆（Scophthalmus maximus）的弧菌病診斷，并利用ELISA技術(shù)，驗證了該抗原芯片用于弧菌病診斷的準(zhǔn)確性。本研究為魚類弧菌病的血清學(xué)診斷提供了有力工具。

Construction of Serological Diagnostic Chip for Pathogenic Vibrio in Fish

The antiserum samples of Paralichthys olivaceus were prepared respectively against 6 kinds of pathogenic vibrios，including Vibrio ichthyoenteri，V. anguillarum，V. parahaemolyticus，V. alginolyticus，V. fluvialis and V. harveyi. The outer membrane protein，flagellum protein，extracellular products and whole-cell bacterial protein of the strains were extracted and purified，then the immune reactions between the antigen components and the antiserum samples were analyzed，it was found that，for all Vibrios，the immune reaction between its antigen and its own antiserum was the strongest，besides，there were also cross-reactions in different extent between the antiserum with the antigen of other Vibrios. Further，the antigen components of the 6 strains were fixed on the chip carrier to form an antigen microarray so as to construct the serological diagnostic antigen chip，the method for interpreting detection results was determined by using monoclonal antibody 2D8 of IgM marked with horseradish peroxidase as the detection antibody；the antigen chip was applied to diagnose vibriosis in Paralichthys olivaceus and Scophthalmus maximus，and its accuracy was verified by ELISA. As a conclusion，a useful tool was provided for serological diagnosis of fish vibriosis.

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