广东海洋大学学报2025，Vol.45Issue(2)：52-58,7.DOI:10.3969/j.issn.1673-9159.2025.02.007
对虾肝肠胞虫LAMP-LFD检测方法的建立及应用
Establishment and Application of LAMP-LFD Method for Detection of Enterocytozoon hepatopenaei
摘要
Abstract
[Objective]Loop-mediated isothermal amplification(LAMP)technology in combination with the lateral flow dipstick(LFD)was used to detect Enterocytozoon hepatopenaei(EHP)in shrimp,and to provide technical support for early disease surveillance in shrimp farms.[Method]Four specific primers(upstream primer FIP was labeled with biotin)and one fluorescein isothiocyanate probe were designed for 18S rDNA sequence of E.hepatopenaei.The biotin-labeled LAMP reaction was performed using primers,and the product was hybridized to the FITC-labeled probe,and the hybridization product was displayed on the LFD.The specificity,sensitivity and clinical application of the method were tested.[Result]The optimized reaction condition for LAMP was running at 62 ℃ for 15 min,probe hybridization was added for 5 min,and then reaction products were detected by LFD strip as the indicator.The established LAMP-FLD method has strong specificity and high sensitivity,which is 100-fold more sensitive than that of ordinary PCR methods.The LAMP-LFD method established in this study was employed to detect E.hepatopenaei,white spot syndrome virus,decapod iridescent virus 1,and Vibrio parahaemolyticus in both artificially infected Litopenaeus vannamei hepatopancreatic tissues and naturally occurring L.vannamei samples suspected of E.hepatopenaei,infection.The results showed that the LAMP-LFD method could 100%accurately detect E.hepatopenaei in these samples.[Conclusion]LAMP-LFD established in this study for the visual detection of E.hepatopenaei.with strong specificity,high sensitivity,simple operation,low cost and less time-consuming,is suitable for the on-site and immediate detection of E.hepatopenaei infections in shrimp farms.关键词
对虾肝肠胞虫/环介导等温扩增技术/横向流动试纸条/凡纳滨对虾Key words
Enterocytozoon hepatopenaei/loop-mediated isothermal amplification/lateral flow dipstick/Litopenaeus vannamei分类
水产学引用本文复制引用
李文泽,侯晓勇,杨世平,黄郁葱,简纪常,蔡双虎..对虾肝肠胞虫LAMP-LFD检测方法的建立及应用[J].广东海洋大学学报,2025,45(2):52-58,7.基金项目
广东省现代农业产业技术体系创新团队建设项目(2024CXTD25) （2024CXTD25）
