Nervous necrosis virus (NNV) is an infectious pathogen, characterized by rapid infection and high mortality, and is commonly encountered in the aquaculture industry. There is an urgent need to develop effective antiviral agents against NNV to establish targeted therapeutic strategies and control viral spread, thereby reducing economic losses. Screening for potent antiviral compounds is critical for the development of highly effective anti-NNV drugs. In this study, a target-activatable detection probe (TAA-TNA1c) was constructed based on the previously screened aptamer TNA1c, which specifically recognizes NNV-infected cells. This probe was employed for high-throughput screening of anti-NNV compounds. The results demonstrated that TAA-TNA1c specifically recognized NNV-infected cells, with positive signals detectable as early as 6 h post-infection at a viral concentration of 0.2 μL/100 μL, indicating its ability to rapidly and efficiently identify NNV infection. Furthermore, antiviral drug screening using this probe yielded results consistent with those obtained by RT-qPCR. Notably, the entire screening process was completed within 2 h, highlighting the potential of TAA-TNA1c as a rapid screening probe for anti-NNV drugs. This study provides essential experimental data and a theoretical foundation for the development of antiviral therapeutics. Therefore, there is an urgent need to develop effective antiviral drugs against NNV, which is crucial for targeted treatment strategies, pathogen control, and loss reduction. Screening of effective antiviral active compounds, namely drug precursors, is key to developing highly efficient drugs against NNV. Reverse transcription-quantitative real-time PCR (RT-qPCR) is widely applied to screen and evaluate various substances to prevent and control virus infection. However, RT-qPCR is a cumbersome procedure and not suitable for high-throughput rapid screening. The aptamer TNA1c, which specifically binds to NNV-infected cells, was used to construct a target-driven activated aptamer probe (TAA). Then, the TAA probe was applied to establish a high-throughput screening (TAA-HTS) method for efficient evaluation of substances against NNV infection. TAA-HTS technology achieved rapid, sensitive, and specific screening of bioactive substances with significant anti-NNV effects. As compared to commonly used analytical methods, such as RT-qPCR, TAA-HTS has the advantages of easy operation and high sensitivity and specificity. The findings of this study provide data support and a theoretical basis for the development of effective antivirus preparations.
Liu et al. (Sun,) studied this question.