Establishment and application of RT-RAA-LDF method for rapid detection of por-cine rotavirus
10.16303/j.cnki.1005-4545.2024.12.05
- VernacularTitle:猪轮状病毒RT-RAA-LFD快速诊断方法的建立及应用
- Author:
Xiaoying LUO
1
;
Fuzhong LIN
;
Qiujuan LIN
;
Yanqin ZENG
;
Zhen CHEN
;
Wenhao SHI
Author Information
1. 福州市农业科学研究所,福建 福州 350018
- Publication Type:Journal Article
- Keywords:
porcine rotavirus;
recombinase aided amplification;
lateral flow dipstick;
detection meth-od
- From:
Chinese Journal of Veterinary Science
2024;44(12):2534-2539
- CountryChina
- Language:Chinese
-
Abstract:
To rapidly detect porcine rotavirus(PoRV),a one-step detection method was developed based on recombinase-aided amplification(RAA)combined with lateral flow dipstick(LFD)tech-nology.Specific primers and probes were designed according to the conserved sequence of the PoRV VP6 gene,and the amplification system was optimized.The specificity,sensitivity,and re-peatability of the method were thoroughly tested,and its clinical application was evaluated.The re-sults demonstrated that the method could amplify PoRV nucleic acid at 37 ℃ in just 15 min with a minimal detection limit of 22.3 copies/μL.No cross-reactivity was observed with other common diarrhea viruses.The total coincidence rate between RT-RAA-LFD and RT-PCR was 97.0％,with a Kappa coefficient of 0.93(K＞0.75).In summary,the RT-RAA-LFD detection method estab-lished in this study exhibits excellent specificity,sensitivity,and stability,characterized by rapid detection speed and visualization,making it suitable for on-site rapid detection.This method pro-vides a novel technical approach for the rapid diagnosis and epidemiological investigation of PoRV.
