生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (2): 127-.doi: 10.3969/j.issn.2095-1736.2023.02.127

• 教学研究 • 上一篇    

可视化测定病毒滴度及抗病毒药物药效的方法优化

唐　崎， 曹苑苑， 张欣雨， 朋王平， 芦宝静， 黄升海   

  1. 安徽医科大学 基础医学院， 合肥 230032
  • 出版日期:2023-04-18 发布日期:2023-04-18
  • 通讯作者: 黄升海，博士，教授，研究方向为病原微生物致病机制研究，E-mail:shhuang@ahmu.edu.cn
  • 作者简介:唐崎，博士，校聘副教授，研究方向为抗病毒药物研发及其机制研究，E-mail:tangqi@ahmu.edu.cn
  • 基金资助:
    国家自然基金青年基金项目（82102374）；安徽省省级质量工程项目（2019zyrc017，2021jcxkpy013，2020mooc200）；省级重大教研项目（2020jyxm0864）；省级创新训练项目（S202210366074）

Optimization of the methods for visual determination of viral titer and efficacy of antiviral drugs

TANG Qi, CAO Yuanyuan, ZHANG Xinyu, PENG Wangping, LU Baojing, HUANG Shenghai   

  1. School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China
  • Online:2023-04-18 Published:2023-04-18

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摘要： 实验课程教学是培养学生创新能力的重要抓手，在生物医学专业的细胞培养课程中，测定病毒滴度和抗病毒药物药效这一综合实验是其重要内容，但是传统的病毒空斑实验在教学中存在诸多问题。首先通过优化传统空斑实验步骤，提高实验效率；其次通过整合空斑实验和药物实验，简化抗病毒药物实验方法；通过定量且可视化的测定病毒滴度和抗病毒药物药效，提高学生的实验技能和科研兴趣。此外，通过前沿技术荧光酶联免疫斑点（Fluo-ELISPOT），可视化和定量检测病毒噬斑（FFU），克服传统空斑实验不能检测致细胞病变效应（CPE）不显著病毒的滴度和抗病毒药物药效，为抗病毒药物研发提供新的实验手段和新的见解。优化的医学生物学实验课程体系提高了细胞课程教学质量，推动了学生实验课程的改革和创新。

关键词: 抗病毒药物, 空斑实验, 荧光酶联免疫斑点, 实验教学改革, 微生物学

Abstract: Experimental course teaching is an important measure to cultivate students’ innovative abilities. In the cell culture course of biomedical majors, the comprehensive experiment of determining viral titer and antiviral drug efficacy is important content. However, there are many problems in the course of traditional plaque experiments. In this paper, the experimental efficiency was first improved by optimizing the traditional steps of plaque assays. By integrating plaque assay and drug assay, the experimental methods of determining the effect of antiviral drugs were simplified. And the experimental skills and scientific interest of students were enhanced through the quantitative and visual determination of viral titers and antiviral efficacy. In addition, the traditional plaque assays neither detect titers of the viruses without significant cytopathic effects (CPE), nor test the activity of antiviral agents to these viruses. In this paper, to address these problems, a cutting-edge technology-fluorescent enzyme-linked immune spots (Fluo-ELISPOT) was applied to visualize and quantify viral plaques and to determine viral titers and efficacy of antiviral drugs, which provided new experimental techniques and new insights for the research and development of antiviral drugs. It also made a positive attempt to optimize the experimental teaching system of medical biology and improve the quality of cell experimental teaching, and further promoted the reform and innovation of student experimental teaching.

Key words: antiviral agents, plaque assay, Fluo-ELISPOT, experimental teaching reform, microbiology

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