分子生物学
IVD分子诊断
细胞培养与分析
蛋白研究
细胞因子
重组蛋白
抗体
高通量测序建库
病原检测UCF系列
生物医药
工具酶
抑制剂激活剂与常用试剂
仪器
耗材

Developing a robust method integrating with selective membrane-based preconcentration and signal amplification for field virus detection

Ruohong Chen, Xingni Chen, Hongtao Liu, Ling Fang, Baowei Chen, Tiangang Luan

Journal:ANALYTICA CHIMICA ACTA

IF:6.91

DOI:10.1016/j.aca.2022.340360

PMID:36156222

Published:2022-09-08

research field:分子生物学比较免疫学兽医学免疫学传染病学

Abstract

Infectious diseases caused by viruses have attracted global concern owing to their rapid spread and catastrophic consequences. Therefore, developing fast and reliable on-site virus detection methods is essential for the prevention and treatment of virus-related diseases. In this study, immunoassays on a membrane, combining virus preconcentration with nanoparticle-based signal amplification, were used to realize the rapid and accurate visual detection of viruses. The biotin–streptavidin scaffolds for target virus preconcentration were established on a membrane, and subsequently a Zika aptamer (Apt) immobilized on the membrane recognized and captured the nonstructural protein 1 of Zika virus (ZIKV-NS1). The probe for detection was synthesized by conjugating the Zika Apt with a high level of horseradish peroxidase on gold nanoparticles. The ZIKV-loaded membrane was incubated with the probes, and the viral signal was amplified as the signal of horseradish peroxidase. In the presence of 3,3,5′,5′-tetramethyl benzidine and hydrogen peroxide, the green color of the probe-coated membrane indicated the level of ZIKV-NS1. Our developed method could reach a detection limit of 5 ng mL −1 , and the whole procedure could be completed within 1 h. Analyses of rabbit serum and environmental water samples demonstrated that an immunoassay-based approach on the membrane could accurately determine the level of ZIKV-NS1 against the complicated matrix. Our results suggest that this virus detection method has a high potential for application in clinical and environmental settings.

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