开发用于快速检测对虾急性肝胰腺坏死病致病性 PirAB 毒素的侧流免疫测定条

Tran Van Hieu, Duong Ngoc Diem, Nguyen Phuoc Khai Hoan, Do Thi Kim Yen, Huynh Tuan Binh, Mai Hoang Thuy Dung, Nguyen Thi Nguyet Thu, T. L. Thuoc
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摘要

急性肝胰腺坏死病(AHPND)是目前全球对虾养殖场中发病率最高的疾病,快速、特异的检测方法对改善疾病管理至关重要,因此设计了一种侧流免疫分析法(LFIA)来检测对虾中的 PirAvp 和 PirBvp 毒素。本研究开发了基于多克隆抗体的侧流免疫分析条。使用副溶血性弧菌(VPAHPND)的重组 PirAvp 和 PirBvp 类毒素蛋白对兔子进行免疫,第三次加强免疫后抗体滴度最高。这种多克隆兔抗血清经纯化后用于开发检测 AHPND 病原体的 LFIA 试纸。该试纸可在 15 分钟内肉眼检测出 125 ng/mL 以下的 PirAvp 和 PirBvp 毒素。与 VPnon-AHPND 没有交叉反应。此外,LFIA 的灵敏度和特异性分别为 94.0% 和 98.0%。该 LFIA 试剂条可作为农民和技术人员就地使用的便捷工具。® 2023 科技期刊 - NTTU
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Development of lateral flow immunoassay strip for rapid detection of acute hepatopancreatic necrosis disease-causing PirAB toxins in shrimp
Acute hepatopancreatic necrosis disease (AHPND) is currently the most affected disease of shrimp farms worlwide As rapid and specific detection methods are essential to improve disease management, a lateral flow immunoassay (LFIA) was designed for the detection of PirAvp and PirBvp toxins in shrimp. In this study, polyclonal antibody based lateral flow immunoassay (LFIA) strip has been developed. The recombinant PirAvp and PirBvp toxin‐like proteins of Vibrio parahaemolyticus (VPAHPND) was used to immunize rabbits with the highest antibody titre after the third booster. This polyclonal rabbit antiserum was purified and used to develop a LFIA strip for the detection of AHPND pathogen. This test strip could detect PirAvp and PirBvp toxins up to 125 ng/mL by naked eye within 15 min. No cross-reactivity was observed with VPnon-AHPND. Furthermore, the sensitivity and specificity of LFIA were 94.0% and 98.0%, respectively. This LFIA strip could be used as a handy tool by the farmers and technicians in situ. ® 2023 Journal of Science and Technology - NTTU
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