中国动物检疫2023，Vol.40Issue(12)：91-97,7.DOI:10.3969/j.issn.1005-944X.2023.12.018
致急性肝胰腺坏死病副溶血弧菌RAA-LFD快检方法的建立
Development of a RAA-LFD Assay for Rapid Detection of Vibrio Parahaemolyticus Resulting in Acute Hepatopancreas Necrosis Disease
摘要
Abstract
In order to develop a simple and rapid on-site detection method for Vibrio parahaemolyticus resulting in acute hepatopancreas necrosis disease(VpAHPND),RAA-nfo specific primers and probe were designed based on the conserved sequence of pirB gene of the VpAHPND pVA1 plasmid,and a RAA-LFD method was established.The method,with strong specificity,could detect positive nucleic acids of specific virulence strains of VpAHPND,but failed to crossly react with pathogenic nucleic acids such as non-virulence strain of Vibrio parahaemolyticus,Vibrio cholerae,Shigella,Salmonella,white spot syndrome virus(WSSV),infectious subcutaneous and hematopoietic necrosis virus(IHHNV),decapod iridescent virus 1(DIV1)and enterocytozoon hepatopenaei(EHP);it was with high sensitivity,could detect the minimum bacterial cells at 3.8×102 CFU/mL,and plasmids at 1.17×101 copies/μL;the results could be determined after reacting at 37℃for 20 min and color development for 5 min by LFD,shortening the detection time;besides,good repeatability and reliability were also observed,58 strains of Vibrio parahaemolyticus were parallelly tested by the established method and the qPCR recommended by WOAH,with 100%coincidence rate.In conclusion,the established method was specific,sensitive,rapid and simple with visual results,and appropriate for on-site rapid detection of VpAHPND.关键词
急性肝胰腺坏死病/副溶血弧菌/重组酶介导扩增技术/侧流层析技术Key words
acute hepatopancreas necrosis disease/Vibrio parahaemolyticus/recombinase-aid amplification/lateral flow test分类
农业科技引用本文复制引用
田飞焱,顾泽茂,徐节华,吴葵,黄海莉,孟霞,裴建明,刘文珍,张锦波,李小勇,周文华..致急性肝胰腺坏死病副溶血弧菌RAA-LFD快检方法的建立[J].中国动物检疫,2023,40(12):91-97,7.基金项目
江西省农牧渔业科研指导性课题项目(赣农厅办函[2021]36号-序号29,赣农厅办函[2023]14号-序号32) （赣农厅办函[2021]36号-序号29,赣农厅办函[2023]14号-序号32）
江西省重点研发计划项目(20202BBG73031) （20202BBG73031）
