中国动物检疫2024，Vol.41Issue(9)：111-117,7.DOI:10.3969/j.issn.1005-944X.2024.09.020
赤羽病病毒荧光定量RT-PCR检测方法的建立及应用
Establishment and Application of a Fluorescence Quantitative RT-PCR for Akabane Disease Virus
摘要
Abstract
In order to establish a rapid and sensitive nucleic acid detection method for Akabane disease virus(AKAV),a fluorescence quantitative RT-PCR assay for AKAV was developed targeting at AKAV S gene,through designing specific primers and Taq Man probe and optimizing reaction system and conditions,followed by evaluation on its sensitivity,specificity and reproducibility,then 80 clinical bovine and ovine blood samples were simultaneously detected using the established method and that recommended by AKAV quarantine industry standard.The results revealed that the established method was highly sensitive with the lowest detection limit of 13.4 copies/μL;good specificity was observed without cross-reaction with foot-and-mouth disease virus(FMDV),bovine coronavirus(BCV),lumpy skin disease virus(LSDV),bovine viral diarrhoea virus(BVDV),infectious bovine rhinotracheitis virus(IBRV),bluetongue virus(BTV)and bovine leukaemia virus(BLV);good reproducibility was represented by intra-and-inter group coefficients of variation of 0.93%-2.47%and 1.25%-2.76%,respectively;and 21 out of 80 clinical samples were positive as detected by the method,98.75%coincident with that by the recommended method by AKAV quarantine industry standard.In conclusion,the established method,with high sensitivity,good specificity and reproducibility,was appropriate to detect Akabane disease(AKD),technically supporting the development of pathogenetic monitoring and diagnostic technology standards for AKAV.关键词
赤羽病病毒/实时荧光定量RT-PCR/方法建立/初步应用Key words
AKAV/real-time fluorescence quantitative RT-PCR/development of method/application分类
农业科技引用本文复制引用
李超,王素春,隋金钰,潘俊慧,魏世萌,祁倩,周凯钰太,王楷宬..赤羽病病毒荧光定量RT-PCR检测方法的建立及应用[J].中国动物检疫,2024,41(9):111-117,7.基金项目
国家重点研发计划项目(2021YFD1800500) （2021YFD1800500）
