title: Selection and characterization of novel DNA aptamers specifically recognized by Singapore grouper iridovirus-infected fish cells.
abstract: Singapore grouper iridovirus (SGIV) is a major viral pathogen of grouper aquaculture, and has caused heavy economic losses in China and South-east Asia. In this study, we generated four ssDNA aptamers against SGIV-infected grouper spleen (GS) cells using SELEX (systematic evolution of ligands by exponential enrichment) technology. Four aptamers exhibited high affinity to SGIV-infected GS cells, in particular the Q2 aptamer. Q2 had a binding affinity of 12.09 nM, the highest of the four aptamers. These aptamers also recognized SGIV-infected tissues with high levels of specificity. Protease treatment and flow cytometry analysis of SGIV-infected cells revealed that the target molecules of the Q3, Q4 and Q5 aptamers were trypsin-sensitive proteins, whilst the target molecules of Q2 might be membrane lipids or surface proteins that were not trypsin-sensitive. The generated aptamers appeared to inhibit SGIV infection in vitro. Aptamer Q2 conferred the highest levels of protection against SGIV and was able to inhibit SGIV infection in a dose-dependent manner. In addition, Q2 was efficiently internalized by SGIV-infected GS cells and localized at the viral assembly sites. Our results demonstrated that the four novel aptamers we generated were specific for SGIV-infected cells and could potentially be applied as rapid molecular diagnostic test reagents or therapeutic drugs targeting SGIV.
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.|University of Chinese Academy of Sciences, 19 Yuquan Road, Beijing 100049, PR China.
authors: Pengfei
authors: P
authors: Li
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.
authors: Shina
authors: S
authors: Wei
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.|University of Chinese Academy of Sciences, 19 Yuquan Road, Beijing 100049, PR China.
authors: Lingli
authors: L
authors: Zhou
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.
authors: Min
authors: M
authors: Yang
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.|University of Chinese Academy of Sciences, 19 Yuquan Road, Beijing 100049, PR China.
authors: Yepin
authors: Y
authors: Yu
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.
authors: Jingguang
authors: J
authors: Wei
authors: Analytical and Testing Center, Beijing Normal University, Xinjiekouwai Street, Beijing 100875, PR China.
authors: Guohua
authors: G
authors: Jiang
authors: Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.|University of Chinese Academy of Sciences, 19 Yuquan Road, Beijing 100049, PR China.
authors: Qiwei
authors: Q
authors: Qin
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000235:genetics*; D004266:DNA Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D005393:Fish Diseases / Q000175:diagnosis / Q000821:virology*; D005399:Fishes; D017995:Ranavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D052156:SELEX Aptamer Technique / Q000379:methods*
chemical_list: D052157:Aptamers, Nucleotide