Acta Virol. 2022;66(3):275-280. doi: 10.4149/av_2022_307.
Standard assays based on ELISA and RT-PCR have been widely used to detect flaviviral infections, including the Zika virus (Zika). Despite their simple, unique, and sensitive features, RT-PCR and ELISA-based assays cannot meet the requirements of high-throughput screening of bulk samples during an outbreak. Several research groups around the world are working on the development of rapid, multiplex, and sensitive assays to overcome the limitations of standard assays used in viral detection. Recent advances in flow cytometry have led to remarkable progress in its use as a basic analysis tool in laboratories. Here, we used the advantages of flow cytometry to develop a Zika detection assay using recombinant Zika envelope (E) protein. The E protein-based flow cytometry assay was able to detect anti-Zika E antibodies from Zika-infected patients, Zika-infected mice, and mice immunized with recombinant Zika E protein. We report the development of the first flow cytometry-based diagnostic assay that can be used for Zika detection. Its rapid turnaround time and ability to detect antibodies from Zika-infected patients can be used to improve the diagnostic accuracy of Zika detection. Keywords: Flavivirus; Zika virus; E protein; NS-1 protein; flow cytometry; ELISA; RT-PCR.
基于 ELISA 和 RT-PCR 的标准检测方法已被广泛用于检测黄病毒感染,包括寨卡病毒(Zika)。尽管它们具有简单、独特和敏感的特点,但 RT-PCR 和 ELISA 为基础的检测方法无法满足在疫情爆发时对大量样本进行高通量筛选的要求。世界各地的几个研究小组正在研究开发快速、多重和敏感的检测方法,以克服用于病毒检测的标准检测方法的局限性。流式细胞术的最新进展使其在实验室作为基本分析工具的使用方面取得了显著进展。在这里,我们利用流式细胞术的优势,使用重组寨卡包膜(E)蛋白开发了一种寨卡检测方法。基于 E 蛋白的流式细胞术检测方法能够检测到寨卡病毒感染患者、感染寨卡病毒的小鼠以及用重组寨卡 E 蛋白免疫的小鼠中的抗寨卡 E 抗体。我们报告了第一个基于流式细胞术的诊断检测方法的开发,该方法可用于寨卡检测。其快速的周转时间和检测寨卡病毒感染患者抗体的能力可用于提高寨卡检测的诊断准确性。关键词:黄病毒;寨卡病毒;E 蛋白;NS-1 蛋白;流式细胞术;ELISA;RT-PCR。