文献类型：期刊文献

英文题名：Sensitivity improvement of rapid Vibrio harveyi detection with an enhanced chemiluminescent-based dot blot

作者：Li, H.[1,2];Xiao, J.[1,2];Zhou, Y.[1,2];Wang, Q.[1,2,3];Zhang, Y.[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Engn Res Ctr Maricultured Anim Vaccines, Shanghai, Peoples R China;[3]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai, Peoples R China

年份：2017

卷号：65

期号：3

起止页码：206

外文期刊名：LETTERS IN APPLIED MICROBIOLOGY

收录：;EI(收录号：20231113726696);WOS:【SCI-EXPANDED(收录号:WOS:000407677500003)】；

基金：We thank Dr Zhang Xiaohua (Ocean University of China, China) and Dr Zhao Lanmo (Institute of Oceanology, Chinese Academy of Sciences, China) for kindly sending strains used in this study. This work was supported by grants from the Ministry of Agriculture of China (Nos. CARS-50 and nyhyzx-201303047), and the Project of Marine Public Welfare Profession (No. 201405003-2).

语种：英文

外文关键词：dot blot; enhanced chemiluminescent; hatcheries; monoclonal antibodies; rapid detection; Vibrio harveyi

摘要：Vibrio harveyi is an opportunistic pathogen in seawater and can cause severe vibriosis. It is prevalent in hatcheries worldwide and can lead to severe economic losses. Therefore, there is an urgent need to develop a rapid detection method for monitoring this pathogen. In this study, to increase the detection sensitivity of our assay with monoclonal antibodies (Mabs) against V.harveyi, the conditions of the dot blot assay were optimized, and enhanced chemiluminescent (ECL) substrate replaced the traditional tetramethylbenzidine (TMB) substrate. Based on the optimization results, an ECL-based novel dot blot assay was developed for the rapid and sensitive detection of V. harveyi. Compared with the traditional dot blot assay, the incubation time was shortened from 8 to 2h. The limit of detection (LOD) for V. harveyi was 2x10(5)CFU per ml (10(3)CFU per spot) in pure bacterial suspension, which was 50-fold more sensitive than the traditional dot blot assay (1x10(7)CFU per ml). Furthermore, when compared with indirect ELISA, the dot blot assay showed approximately 1000-fold higher sensitivity (CFU/CFU). After the test sample was pre-enriched in turbot homogenates for 6h before the dot blot analysis, the LOD for V. harveyi was 10CFU per ml. Significance and Impact of the StudyVibrio harveyi is one of the most opportunistic pathogens that can cause high mortality in hatcheries worldwide. To detect this pathogen, a novel dot blot based on enhanced chemiluminescent (ECL) has been established. This ECL-based dot blot was found to be more sensitive and rapid for V. harveyi detection than traditional dot blot, and this technology is recommended as an applied protocol for monitoring V. harveyi in seawater to reduce economic losses.