中国动物检疫2025，Vol.42Issue(2)：100-107,8.DOI:10.3969/j.issn.1005-944X.2025.02.017
十足目虹彩病毒1 RAA-LFD快检方法建立与应用
Establishment and Application of RAA-LFD Assay for Decapod Iridescent Virus 1
摘要
Abstract
In order to develop an on-site assay for Decapod iridescent virus 1(DIV1),specific primers and probes for RAA-nfo were designed based on the conserved sequence of the gene encoding DIV1 ATPase to develop RAA-LFD assay for on-site detection of DIV1 in combination with lateral flow dipstick(LFD)and through optimizing the conditions of reaction time and temperature.The reaction could be completed at 37℃for 20 min,and the results could be interpreted after 5 minutes of LFD coloration;no cross-reaction with other shrimp pathogens was observed,such as Aeromonas hydrophophilus,Citrobcter freundii,white spot syndrome virus(WSSV),infectious hypodermal and haematopoietic necrosis virus(IHHNV),entamoeba hepatica of shrimp(EHP)and Vibrio parahaemolyticus(VpAHPND),while only DIV1 was tested positive;the minimum detection limit was 5.25×101 copies/reaction;and the results of intra-and inter-group reproducibility tests were good.60 Procambarus clarkii samples were detected using the assay in parallel with the reported nested PCR and qPCR,the results of the assay were consistent with those using the nested PCR,and the sensitivity and specificity were 83.3%and 100%,respectively,compared with the qPCR.In conclusion,the established assay was rapid,accessible,highly sensitive and specific,with good reproducibility and reliability,and appropriate for on-site detection of clinical samples.关键词
十足目虹彩病毒1/重组酶介导扩增技术/侧流层析技术/恒温扩增/快速检测Key words
DIV1/recombinase-aid amplification/LFD/isothermal amplification/rapid detection分类
农业科技引用本文复制引用
田飞焱,吴众怡,徐节华,黄海莉,裴建明,孟霞,刘文珍,周文华,谢世红..十足目虹彩病毒1 RAA-LFD快检方法建立与应用[J].中国动物检疫,2025,42(2):100-107,8.基金项目
江西省大宗淡水鱼产业技术体系病害防控岗项目(2025-2027年) （2025-2027年）
江西省农牧渔业科研指导性课题项目(2021-29,2023-32) （2021-29,2023-32）
