College of Chemistry, Sichuan University, Chengdu, 610064, PR China.
College of Chemistry, Sichuan University, Chengdu, 610064, PR China.
Anal Chim Acta. 2021 Aug 1;1171:338645. doi: 10.1016/j.aca.2021.338645. Epub 2021 May 17.
Rapid and low-cost diagnosis of multiple infectious diseases is of great significance especially in densely populated or resource-constrained settings. Herein, we developed a one-step fast and label-free imaging array for multiplexed detection of trace avian influenza virus (AIV) DNA biomarkers. By designing a series of specific and efficient catalytic hairpin assembly (CHA) amplification reactions and utilizing thioflavin T, a specific G-quadruplex fluorescence probe, three subtypes of AIV DNA biomarkers (H1N1, H7N9 and H5N1) were simultaneously and quickly detected within only 20 min, which just needed a small reagent volume of 50 μL and a smartphone instead of a spectrometer. With the combination of fluorescence imaging output and grey-level analysis, the array sensor can be on-site with the limit of detection of 136 pM, 141 pM and 129 pM for H1N1, H7N9 and H5N1, respectively. The imaging array also displayed good mismatch discrimination, excellent anti-interference, and real sample application. In view of its advantages of fast detection, low cost and multiplexed analysis, the imaging array is expected to have potential applications for early infectious disease diagnosis in resource-constrained settings.
快速且低成本的多种传染病诊断具有重要意义，尤其是在人口密集或资源有限的环境中。在此，我们开发了一种一步式快速、无标记的成像阵列，用于同时检测痕量禽流感病毒（AIV）DNA 生物标志物。通过设计一系列特异性和高效的催化发夹组装（CHA）扩增反应，并利用硫堇 T（一种特异性 G-四链体荧光探针），我们在 20 分钟内即可同时快速检测三种 AIV DNA 生物标志物（H1N1、H7N9 和 H5N1），仅需 50 μL 的小试剂体积和智能手机即可，而无需光谱仪。通过荧光成像输出和灰度分析相结合，该阵列传感器可现场检测，H1N1、H7N9 和 H5N1 的检测限分别为 136 pM、141 pM 和 129 pM。该成像阵列还显示出良好的错配区分、出色的抗干扰能力和实际样本应用。鉴于其快速检测、低成本和多重分析的优势，该成像阵列有望在资源有限的环境中用于早期传染病诊断。
