College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao, P. R. China.
Samarkand State University named after Sh. Rashidov, Samarkand, Uzbekistan.
Anal Methods. 2024 Aug 29;16(34):5777-5784. doi: 10.1039/d4ay01010h.
Sensitive, convenient and rapid detection and subtyping of influenza viruses are crucial for timely treatment and management of infected people. Compared with antigen detection, nucleic acid detection has higher specificity and can shorten the detection window. Hence, in this work, we improved the lateral flow assay (LFA, one of the most promising user-friendly and on-site methods) to achieve detection and subtyping of H1N1, H3N2 and H9N2 influenza virus nucleic acids. Firstly, the antigen-antibody recognition mode was transformed into a nucleic acid hybridization reaction. Secondly, FeO-Au heterodimer nanoparticles were prepared to replace frequently used Au nanoparticles to obtain better coloration. Thirdly, four lines were arranged on the LFA strip, which were three test (T) lines and one control (C) line. Three T lines were respectively sprayed by the DNA sequences complementary to one end of H1N1, H3N2 and H9N2 influenza virus nucleic acids, while FeO-Au nanoparticles were respectively coupled with the DNA sequences complementary to the other end of H1N1, H3N2 and H9N2 nucleic acids to construct three kinds of probes. The C line was sprayed by the complementary sequences to the DNAs on all three kinds of probes. In the detection, by hybridization reaction, the probes were combined with their target nucleic acids which were captured by the corresponding T lines to form color bands. Finally, according to the position of the color bands and their grey intensity, simultaneous qualitative and semi-quantitative detection of the three influenza virus nucleic acids was realized. The detection results showed that this multi-channel LFA had good specificity, and there was no significant cross reactivity among the three subtypes of influenza viruses. The simultaneous detection achieved comparable detection limits with individual detections. Therefore, this multi-channel LFA had good application potential for sensitive and rapid detection and subtyping of influenza viruses.
灵敏、便捷、快速的流感病毒检测和亚型鉴定对于感染人群的及时治疗和管理至关重要。与抗原检测相比，核酸检测具有更高的特异性，并能缩短检测窗口期。因此，本工作旨在改进侧向流检测（LFA，一种最有前途的用户友好型和现场检测方法），以实现对 H1N1、H3N2 和 H9N2 流感病毒核酸的检测和亚型鉴定。首先，将抗原-抗体识别模式转变为核酸杂交反应。其次，制备了 FeO-Au 异质二聚纳米粒子以替代常用的金纳米粒子，以获得更好的显色效果。第三，在 LFA 条上排列了四条线，分别是三条检测线（T）和一条控制线（C）。三条 T 线分别喷涂了与 H1N1、H3N2 和 H9N2 流感病毒核酸一端互补的 DNA 序列，同时将 FeO-Au 纳米粒子分别与 H1N1、H3N2 和 H9N2 核酸另一端互补的 DNA 序列偶联，构建了三种探针。C 线喷涂了与三种探针上的 DNA 互补的序列。在检测过程中，通过杂交反应，探针与相应的 T 线捕获的靶核酸结合，形成显色带。最后，根据显色带的位置和灰度强度，实现了三种流感病毒核酸的定性和半定量同时检测。检测结果表明，该多通道 LFA 具有良好的特异性，三种亚型流感病毒之间无明显交叉反应。同时检测的检测限与单独检测相当。因此，该多通道 LFA 具有灵敏、快速检测和鉴定流感病毒的良好应用潜力。
