中国疾病预防控制中心病毒病预防控制所,国家卫生健康委员会生物安全重点实验室;温州医科大学检验医学院生命科学学院,浙江省医学遗传学重点实验室;内蒙古医科大学基础医学院;新乡医学院公共卫生学院;
本研究旨在建立一种快速、通用、高通量的病灶形成测定法（Focus-forming assay, FFA），用于痘苗病毒（Vaccinia virus, VACV）的研究。首先采用经典噬斑染色法与免疫噬斑法对不同VACV代表性毒株（Western Reserve strain, WR; Tiantan strain, VTT; Modified Ankara strain, MVA）在BHK-21细胞和Vero细胞上的噬斑特性进行评估，通过实验条件（包括显色液、非特异性染色、细胞培养板和病毒培养时间）的优化，提高病灶的可视化质量和测定的准确性。并基于改进优化的FFA法研究了VACV在BHK-21细胞和Vero细胞上的生长曲线，同时对FFA法的重复性进行了评估。研究发现BHK-21细胞对VACV的感染更加敏感，适宜于多种VACV滴度测定。实验条件的优化显著提高了病灶的可视化质量和测定的准确性。应用FFA法发现WR、VTT和MVA毒株在BHK-21细胞上能够有效复制，而MVA在Vero细胞上的复制受限，且FFA法在同一实验内具有良好的重复性，变异系数在0.17%到3.50%之间。改进后的FFA法是一种快速、通用、高通量的方法，适用于痘苗病毒滴度的测定，为痘苗病毒的研究和应用提供了可靠的技术支持。
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004508
中图分类号:R373
引用信息:
[1]楚巧鸿,冯霞,李佳等.检测痘苗病毒体外感染的病灶形成方法的优化与应用[J].病毒学报,2024,40(03):492-499.DOI:10.13242/j.cnki.bingduxuebao.004508.
基金信息:
国家重点研发计划（项目号：2023YFD1800405）,题目：抗猴痘药物筛选与阻断技术研究;国家重点研发计划（项目号：2022YFC2303401）,题目：潜在高危新病毒风险识别的新技术体系研究~~