Establishment and application of an RPA-LFD method for detection of Akabane virus
10.16303/j.cnki.1005-4545.2025.08.03
- VernacularTitle:阿卡斑病毒RPA-LFD检测方法的建立及临床应用
- Author:
Jiafu SHANG
1
;
Xuehui ZHOU
;
Yanyan LIU
;
Xia LIU
;
Xingwei NI
;
Tingting XU
;
Zhiguo ZHAO
;
Yan WANG
;
Zhenjun WANG
;
Xiaowei YANG
;
Guangwei ZHAO
Author Information
1. 西南大学动物医学院,重庆 402460
- Publication Type:Journal Article
- Keywords:
Akabane virus;
recombinase polymerase amplification;
lateral flow dipstick;
detection method
- From:
Chinese Journal of Veterinary Science
2025;45(8):1601-1608
- CountryChina
- Language:Chinese
-
Abstract:
To establish a rapid visual detection method for Akabane virus(AKAV)on site,specific primers and probes based on the S fragment of AKAV were designed in this experiment.Corre-sponding groups were added to the primers or probes to fulfil the requirement of the combination of recombinase polymerase amplification(RPA)with lateral flow dipstick(LFD).The reaction temperature and time,concentrations of the primer and probe were optimized to establish the RPA-LFD method for detecting AKAV.After that,the specificity,sensitivity and clinical reliability of the method were evaluated.The results showed that after 20 minutes of reaction at 37 ℃,the test results could be read on LFD paper.There was no cross reaction against blue tongue virus,Pasteurella multocida,bovine infectious rhinotracheitis virus and bovine Mycoplasma bovis,and the detection limit was 2.5 × 100 copies/μL of standard plasmid.Detection of clinical samples showed a consistent results with that by RT-PCR method.These findings indicated that the RPA-LFD method established had the advantages of good specificity,high sensitivity,simple operation and visualization,and could be applied to clinical detection,which provides new technical support for the rapid diagnosis and prevention and control of AKAV.
