Maistriau Marylene, Carletti Tea, Zakaria Mohammad Khalid, Braga Luca, Faoro Valentina, Vasileiadis Vasileios, Marcello Alessandro
Molecular Virology Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy.
High-throughput screening facility, International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy.
Virus Res. 2017 Jun 2;237:1-6. doi: 10.1016/j.virusres.2017.05.004. Epub 2017 May 10.
Virus neutralizing antibodies are critical correlates of protection in vaccine development and are discriminatory in the plaque reduction neutralization test when used for the diagnosis of viral infections. However, neutralization assays are time consuming, labor intensive and highly variable, thus limiting their use. Advances in automated live imaging of cells opened new possibilities for standard virus diagnostic techniques such as neutralization assays. To this end, a reporter cell line based on the translocation of the transcription factor IRF3 in response to infection is proposed. Image acquisition of signal in a microplate format allowed the setup of a rapid, semi-automated and high-throughput fluorescent neutralization test. The study is extended to the live imaging of IRF3 translocations that could potentially cut the time of analysis to few hours. The fluorescent neutralization test is suitable for high-throughput assays and expandable to other viruses of global importance such as Zika virus.
病毒中和抗体是疫苗研发中保护作用的关键相关指标，在用于诊断病毒感染时，在蚀斑减少中和试验中具有鉴别性。然而，中和试验耗时、费力且变化很大，因此限制了它们的应用。细胞自动实时成像技术的进展为诸如中和试验等标准病毒诊断技术开辟了新的可能性。为此，提出了一种基于转录因子IRF3响应感染而发生易位的报告细胞系。以微孔板形式进行信号图像采集，使得能够建立一种快速、半自动且高通量的荧光中和试验。该研究扩展到IRF3易位的实时成像，这有可能将分析时间缩短至数小时。荧光中和试验适用于高通量检测，并且可扩展到其他具有全球重要性的病毒，如寨卡病毒。
