Akpinar Fulya, Yin John
Department of Chemical and Biological Engineering, Systems Biology Theme, Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53706, USA.
J Virol Methods. 2015 Jun 15;218:71-6. doi: 10.1016/j.jviromet.2015.02.006. Epub 2015 Feb 16.
Although transmission electron microscopy (TEM) has historically been the method of choice to estimate concentrations of virus and virus-like particles, these measures can often be time-consuming and labor-intensive to perform. Tunable resistive pulse sensing (TRPS) is an emerging method that applies principles of Coulter counting to nanoscale particles and may provide a simpler and higher-throughput alternative to TEM for the quantitation of virus populations. To assess the performance of TRPS compared to TEM, the samples of polymer spheres at a diameter of 100nm and vesicular stomatitis virus (VSV) were characterized using both techniques. TRPS was able to quantify concentrations down to 10(7)particles/ml, providing nearly 50-fold larger measurement range, and more reproducible counts than TEM. Total-to-infectious particle ratio of VSV populations as measured by TRPS and plaque assay suggested that each VSV particle is infectious. In addition to particle counts, TRPS successfully measured particle size distributions based on hundreds of particles. Such high throughput sustained by TRPS can assist quantitative characterization of virus populations.
尽管传统上透射电子显微镜(TEM)一直是估算病毒和病毒样颗粒浓度的首选方法,但这些检测方法往往既耗时又费力。可调电阻脉冲传感(TRPS)是一种新兴方法,它将库尔特计数原理应用于纳米级颗粒,可能为TEM提供一种更简单、高通量的病毒群体定量替代方法。为了评估TRPS与TEM相比的性能,使用这两种技术对直径为100nm的聚合物球和水疱性口炎病毒(VSV)样本进行了表征。TRPS能够将浓度定量至低至10(7)个颗粒/毫升,测量范围比TEM大近50倍,且计数比TEM更具重复性。通过TRPS和噬斑测定法测量的VSV群体的总感染性颗粒比率表明每个VSV颗粒都具有感染性。除了颗粒计数外,TRPS还基于数百个颗粒成功测量了颗粒大小分布。TRPS所具备的这种高通量能够辅助病毒群体的定量表征。