Department of Chemistry, University of Turku, Henrikinkatu 2, 20500, Turku, Finland.
Institute of Biomedicine, University of Turku, Kiinamyllynkatu 10, 20520, Turku, Finland.
Anal Bioanal Chem. 2022 Jun;414(15):4509-4518. doi: 10.1007/s00216-022-04104-5. Epub 2022 May 17.
Viruses play a major role in modern society and create risks from global pandemics and bioterrorism to challenges in agriculture. Virus infectivity assays and genome copy number determination methods are often used to obtain information on virus preparations used in diagnostics and vaccine development. However, these methods do not provide information on virus particle count. Current methods to measure the number of viral particles are often cumbersome and require highly purified virus preparations and expensive instrumentation. To tackle these problems, we developed a simple and cost-effective time-resolved luminescence-based method for virus particle quantification. This mix-and-measure technique is based on the recognition of the virus particles by an external Eu-peptide probe, providing results on virus count in minutes. The method enables the detection of non-enveloped and enveloped viruses, having over tenfold higher detectability for enveloped, dynamic range from 5E6 to 3E10 vp/mL, than non-enveloped viruses. Multiple non-enveloped and enveloped viruses were used to demonstrate the functionality and robustness of the Protein-Probe method.
病毒在现代社会中扮演着重要的角色,它们会带来从全球大流行和生物恐怖主义到农业挑战等各种风险。病毒感染力测定和基因组拷贝数测定方法通常用于获得用于诊断和疫苗开发的病毒制剂的信息。然而,这些方法并不能提供病毒颗粒数的信息。目前测量病毒颗粒数的方法通常很繁琐,需要高度纯化的病毒制剂和昂贵的仪器。为了解决这些问题,我们开发了一种简单且具有成本效益的基于时间分辨光激化学发光的病毒颗粒定量方法。这种混合和测量技术基于外部 Eu-肽探针对病毒颗粒的识别,可在几分钟内提供病毒计数结果。该方法可检测无包膜和包膜病毒,对包膜病毒的检测灵敏度比非包膜病毒高十倍以上,动态范围为 5E6 到 3E10 vp/mL。我们使用多种非包膜和包膜病毒来证明 Protein-Probe 方法的功能和稳健性。