广东海洋大学学报2024，Vol.44Issue(3)：9-16,8.DOI:10.3969/j.issn.1673-9159.2024.03.002
基于核酸适体的高通量模型筛选抗大口黑鲈虹彩病毒药物
Aptamer-Based High-Throughput Screening Model for Efficient Selection and Evaluation of Medicines against Largemouth Bass Virus Infection
摘要
Abstract
[Objective]In this study,aptamer LBVA1 of Micropterus salmoides iridovirus(Largemouth bass virus,LMBV)was developed with high-throughput screening technology(aptamer LBVA1-based high-throughput screening,LBVA1-AHTS)to rapidly screen effective anti-LMBV drugs.[Method]The probe FAM-LBVA1 was used to detect the fluorescence values of the fathead minnow cells(FHM)challenged by LMBV with multiplicity of infection(MOI)of 0,0.05,0.10,and 0.20,and infection time of 0,6,12,24,and 48 h,respectively,and the infection was verified by RT-qPCR.The basic operation method used nine drugs,including ribavirin,acyclovir,gabapentin,teatoxin,epigallocatechin gallate,etc.,mixed with LMBV at a safe working concentration,and then added to FHM for co-incubation.We used the probe FAM-LBVA1 and RT-qPCR technology to detect LMBV infection in each group,compare the anti-LMBV infection effect of the two methods,and select the effective anti-LMBV drugs.[Result]FAM-LBVA1 detection and RT-qPCR technology showed that the detection effect of LMBV infection was the best when MOI was 0.2 and the infection time was 48 h.The results of screening the anti-LMBV effect of 9 drugs showed that the antiviral effects of 7 drugs,namely ribavirin,gabapentin,amantadine,carbamazepine,tea polyphenols,epigallocatechin gallate and epicatechin gallate,were consistent with the antiviral effects of LBVA1-AHTS and RT-qPCR,of which 3 drugs including tea polyphenols,epigallocatechin gallate and epicatechin gallate had anti-LMBV effect,and the other 4 drugs had no obvious antiviral effect,and the consistency rate of the two methods was nearly 80%.[Conclusion]The established LBVA1-AHTS model facilitates quick,accurate and efficient screening of anti-LMBV drugs,which provides a good theoretical and technical basis for the rapid screening of antiviral drugs.关键词
大口黑鲈虹彩病毒/核酸适体/高通量筛选/抗病毒药物Key words
Largemouth bass virus/aptamer/high-throughput screening/antiviral medicines分类
农业科技引用本文复制引用
张帅帅,陆兰天,杨辉,余庆,李鹏飞,刘明珠,黄静,师德强,陈嘉,柯珂,竺利波,韦云依,牟容丽..基于核酸适体的高通量模型筛选抗大口黑鲈虹彩病毒药物[J].广东海洋大学学报,2024,44(3):9-16,8.基金项目
国家自然科学基金(32373175) （32373175）
广西自然科学基金(2023JJG130007,2022GXNSFBA035521,2022JJA130074) （2023JJG130007,2022GXNSFBA035521,2022JJA130074）
国家重点研发计划(2022YFD2401200) （2022YFD2401200）
国家现代农业产业技术体系广西创新团队项目(nycytxgxcxtd-2021-08-02) （nycytxgxcxtd-2021-08-02）
