Wu Yuting, Liu Shuai, Liu Zhiwei, Liu Bing, Du Bin, Tong Zhaoyang, Xu Jianjie
State Key Laboratory of NBC Protection for Civilian, Beijing, 102205, China.
Synth Syst Biotechnol. 2022 Jul 13;7(4):1066-1072. doi: 10.1016/j.synbio.2022.07.001. eCollection 2022 Dec.
Quantitative analysis is essential for virus research, especially in determining the virus titer. The classical method plaque assay is time-consuming, complex, and difficult for the phages that cannot form apparent plaque on the solid medium. In order to realize rapid and effective detection, a new method combining atomic force microscopy (AFM) observation and mathematical calculation is established. In this research, M13 phages with an appropriate dilution ratio were observed and counted by AFM. Based on the counting results, the titer of M13 phages can be calculated simply through mathematical substitution. Instead of cultivating overnight in plaque assay, this new method can be implemented within a few hours. Moreover, it is a method that can achieve visualization for titer determination and have the potential to determine the phages that fail to form apparent plaque, which is significant in virus quantitative assessment.
定量分析对于病毒研究至关重要，尤其是在确定病毒滴度方面。经典的噬菌斑测定法耗时、复杂，对于无法在固体培养基上形成明显噬菌斑的噬菌体来说操作困难。为了实现快速有效的检测，建立了一种结合原子力显微镜（AFM）观察和数学计算的新方法。在本研究中，通过AFM观察并计数了具有适当稀释比例的M13噬菌体。基于计数结果，只需通过数学代换即可简单计算出M13噬菌体的滴度。与噬菌斑测定法中需要过夜培养不同，这种新方法可以在几小时内完成。此外，这是一种能够实现滴度测定可视化且有潜力测定无法形成明显噬菌斑的噬菌体的方法，在病毒定量评估中具有重要意义。
