Application of polyethylene glycol (PEG) precipitation to concentrate red sea bream iridovirus from seawater = 해수로부터 참돔이리도바이러스 농축을 위한 폴리에틸렌글라이콜(PEG)침전법의 적용

다국어 초록 (Multilingual Abstract)

Red sea bream iridoviral disease (RSIVD), caused by the RSIV and ISKNV genotypes, is listed by the World Organization for Animal Health (OIE) because of its high pathogenicity and impact on over 30 fish species. The detection of RSIV in aquatic water is a vital means of early warning diagnostics, which is helpful for aquatic disease prevention and control in aquaculture. However, detecting viruses in water is challenging because of the low concentration of pathogens in water samples. This study optimized the PEG precipitation process to concentrate RSIV particles from water by combining them with two coagulants: AlCl3 and FeCl3 •6H2O. When PEG was used as a single coagulant, the optimal concentration was 15%(w/v), which exhibited a more stable performance than other concentrations. To enhance precipitation efficiency, experiments with PEG combined with Al or Fe were also conducted. The results showed that in the case of Al, 15% (w,v) PEG combined with Al 0.1 mg/L showed the optimal concentration, whereas 10% (w,v) PEG combined with Fe 0.1 mg/L showed the best precipitation efficiency. The detection limit of this method was approximately 8.0 x 101 RSIV copies/ml of water sample, which is more than 100 times better than that of the method without water sample concentration. The procedure developed in this study is cost-effective, sensitive, and effective for the direct detection of RSIV particles in water using PEG precipitation combined with PCR. Figure legends

목차 (Table of Contents)

• I. Introduction 1
• Ⅱ. Materials and Methods 6
• 1. Virus 6
• 2. Experimental procedure 6
• 2-1. Detection of RSIV particles by conventional PCR 6
• 2-2. Molecular detection limit after artificial seeding with a known RSIV 6
• 2-3. Efficiency of single flocculant on RSIV concentration 7
• 2-4. Efficiency of combined flocculants on RSIV concentration 8
• 2-5. Efficiency of PEG precipitation method 9
• 3. Conventional polymerase chain reaction (PCR) 10
• 4. Statistical analysis 12
• Ⅲ. Results 13
• 1. Detection of RSIV particles by conventional PCR 13
• 2. Molecular detection limit after artificial seeding with a known RSIV 15
• 3. Efficiency of single flocculant on RSIV concentration 17
• 4. Efficiency of combined flocculants on RSIV concentration 20
• 5. Efficiency of PEG precipitation method 25
• Ⅳ. Discussion 27
• Ⅴ. Abstract (Korean). 31
• Ⅵ. References. 32