Department of Chemical Engineering, Michigan Technological University, USA.
Analyst. 2019 Sep 9;144(18):5486-5496. doi: 10.1039/c9an00830f.
Traditional virus detection methods require ligands that bind to either viral capsid proteins or viral nucleic acids. Ligands are typically antibodies or oligonucleotides and they are expensive, have limited chemical stability, and can only detect one specific type of virus at a time. Here, the biochemical surface properties of viruses are exploited for ligand-free, nonspecific virus detection. It has been found that the osmolyte mannitol can preferentially aggregate virus, while leaving proteins in solution. This led to the development of a ligand-free detection of virus using gold nanoparticle (AuNP) aggregation. Porcine parvovirus (PPV) was incubated with AuNPs and aggregation of the PPV-AuNP complex with mannitol was detected by dynamic light scattering (DLS). The lowest detectable concentration of PPV was estimated to be 106 MTT50 per mL, which is lower than standard antibody assays. PPV was also detected when swabbed from a dry surface and in the presence of a protein solution matrix. The enveloped bovine viral diarrhea virus (BVDV) was also detected using mannitol-induced aggregation of BVDV-coated AuNPs. The lowest detectable concentration of BVDV was estimated to be 104 MTT50 per mL. This demonstrates that gold nanoparticle aggregation can detect virus without the use of specific ligands.
传统的病毒检测方法需要与病毒衣壳蛋白或病毒核酸结合的配体。配体通常是抗体或寡核苷酸，它们昂贵、化学稳定性有限，并且一次只能检测一种特定类型的病毒。在这里，利用病毒的生化表面特性进行无配体、非特异性病毒检测。已经发现渗透剂甘露醇可以优先聚集病毒，而将蛋白质留在溶液中。这导致了使用金纳米颗粒（AuNP）聚集来进行无配体的病毒检测的发展。猪细小病毒（PPV）与 AuNP 孵育，并用动态光散射（DLS）检测 PPV-AuNP 复合物与甘露醇的聚集。估计 PPV 的最低可检测浓度为 106 MTT50/mL，低于标准抗体检测。当从干燥表面拭取和存在蛋白质溶液基质时，也可以检测到 PPV。用甘露醇诱导包被 BVDV 的 AuNP 聚集也可以检测到包膜牛病毒性腹泻病毒（BVDV）。估计 BVDV 的最低可检测浓度为 104 MTT50/mL。这表明金纳米颗粒聚集可以在不使用特异性配体的情况下检测病毒。
