A droplet digital PCR assay to detect Chinese rice-field eels rhabdovirus.

Autor: Liu W; Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, China., Zhou Y; Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, China., Jiang N; Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, China., Xu C; Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, China., Zhong Q; College of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang, China., Fan Y; Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, China.
Jazyk: angličtina
Zdroj: Journal of fish diseases [J Fish Dis] 2024 Dec; Vol. 47 (12), pp. e14020. Date of Electronic Publication: 2024 Sep 16.
DOI: 10.1111/jfd.14020
Abstrakt: Chinese rice-field eels rhabdovirus (CrERV) causes haemorrhagic disease in Chinese rice-field eels (Monopterus albus), leading to significant mortality and economic losses. Sensitive detection of CrERV nucleic acids is essential to control the spread of this pathogen and to treat infected individuals. Herein, we developed an efficient and sensitive droplet digital PCR (ddPCR) method to rapidly detect and quantify CrERV. The ddPCR assay optimal conditions were an annealing temperature of 53°C, and primer and probe concentrations of 0.5 and 0.25 μM, respectively. The assay had a diagnostic sensitivity of 0.23 copies/μL, and was highly specific, showing no cross reactivity with other viruses (infectious haematopoietic necrosis virus, grass carp reovirus, spring viremia of carp virus, largemouth bass ranavirus, carp edema virus, Chinese giant salamander iridovirus, and white spot syndrome virus). Real-time quantitative PCR testing of 30 Chinese rice-field eels samples detected CrERV in 7 samples (23.3%), whereas ddPCR detected CrERV in 12 samples (40%), demonstrating its higher sensitivity. Thus, ddPCR represents an advanced method to absolutely quantify CrERV in infected fish with low virus concentrations, providing a valuable tool to manage the spread and impact of CrERV.
(© 2024 John Wiley & Sons Ltd.)
Databáze: MEDLINE
Externí odkaz: