A new method for quantitative analysis of M13 bacteriophage by atomic force microscopy

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...

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Veröffentlicht in:Synthetic and systems biotechnology 2022-12, Vol.7 (4), p.1066-1072
Hauptverfasser: Wu, Yuting, Liu, Shuai, Liu, Zhiwei, Liu, Bing, Du, Bin, Tong, Zhaoyang, Xu, Jianjie
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Sprache:eng
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Zusammenfassung: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.
ISSN:2405-805X
2405-805X
DOI:10.1016/j.synbio.2022.07.001