Abstract
Largemouth bass virus (LMBV) is one of the most devastating viral pathogens in farmed Largemouth bass. Aptamers are novel molecule probes and have been widely applied in the field of efficient therapeutic and diagnostic agents development. LMBV-infected fathead minnow cells (LMBV-FHM) served as target cells in this study, and three DNA aptamers (LBVA1, LBVA2, and LBVA3) were generated against target cells by SELEX technology. The selected aptamers could specifically bind to LMBV-FHM cells, with rather high calculated dissociation constants (Kd) of 890.09, 517.22, and 249.31 nM for aptamers LBVA1, LBVA2, and LBVA3, respectively. Three aptamers displayed efficient antiviral activities in vitro. It indicates that the selected aptamers have great potentials in developing efficient anti-viruses treatments. The targets of aptamers LBVA1, LBVA2, and LBVA3 could be membrane proteins on host cells. The targets of aptamers (LBVA1, LBVA2, and LBVA3) come out on the cells surface at 8, 10, 8 h post-infection. As novel molecular probes for accurate recognition, aptamer LBVA3 could detect LMBV infection in vitro and in vivo, it indicates that the selected aptamers could be applied in the development of rapid detective technologies, which are characterized by high sensitivity, accuracy, and easy operation.
Highlights
Largemouth bass is commonly cultured worldwide, which has extremely economic and aquaculture significance (Wang et al, 2019)
The specific recognition of oligonucleotides library from different selection round targeting Largemouth bass virus (LMBV)-Fathead minnow cells (FHM) cells was characterized by flow cytometry
Compared to rather low fluorescence values on the fish cells in each control group, there were rather high fluorescence values on LMBVinfected fish spleen tissue cells incubated with FAM-LBVA1, FAM-LBVA2, and FAM-LBVA3, respectively (Figure 4C)
Summary
Largemouth bass is commonly cultured worldwide, which has extremely economic and aquaculture significance (Wang et al, 2019). The rapid development of largemouth bass aquaculture has led to frequent outbreaks of various pathogens. Aptamers Targeting LMBV-Infected Cells urgent need for research and development of rapid detection technology and efficient antiviral agents for preventing LMBV infection. The detective assays characterized of easy operation and high sensitivity are helpful to combat LMBV outbreaks in aquaculture. Based on aptamers’ excellent properties, aptamers are widely applied to develop rapid diagnosis technologies, which are characterized of easy operation and high sensitivity (Zhang Y. et al, 2019). DNA aptamers were generated against target LMBV-infected FHM cells by SELEX technology for the first time. The selected aptamers have great potentials in developing efficient anti-virus treatments and rapid detective technologies
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