Department of Bioengineering, University of California, 5121 Engineering V, 420 Westwood Plaza, Los Angeles, CA 90095-1600, USA.
Anal Bioanal Chem. 2010 Dec;398(7-8):2955-61. doi: 10.1007/s00216-010-4213-7. Epub 2010 Sep 24.
Availability of a rapid, accurate, and reliable point-of-care (POC) device for detection of infectious agents and pandemic pathogens, such as swine-origin influenza A (H1N1) virus, is crucial for effective patient management and outbreak prevention. Due to its ease of use, rapid processing, and minimal power and laboratory equipment requirements, the lateral-flow (immuno)assay (LFA) has gained much attention in recent years as a possible solution. However, since the sensitivity of LFA has been shown to be inferior to that of the gold standards of pathogen detection, namely cell culture and real-time PCR, LFA remains an ineffective POC assay for preventing pandemic outbreaks. A practical solution for increasing the sensitivity of LFA is to concentrate the target agent in a solution prior to the detection step. In this study, an aqueous two-phase micellar system comprised of the nonionic surfactant Triton X-114 was investigated for concentrating a model virus, namely bacteriophage M13 (M13), prior to LFA. The volume ratio of the two coexisting micellar phases was manipulated to concentrate M13 in the top, micelle-poor phase. The concentration step effectively improved the M13 detection limit of the assay by tenfold from 5 × 10(8) plaque forming units (pfu)/mL to 5 × 10(7) pfu/mL. In the future, the volume ratio can be further manipulated to yield a greater concentration of a target virus and further decrease the detection limits of the LFA.
对于有效管理患者和预防疫情爆发而言，能够快速、准确、可靠地检测出传染病原体和大流行性病原体（例如猪源流感 A（H1N1）病毒）的即时检测（POC）设备至关重要。由于侧向流动（免疫）检测（LFA）易于使用、处理快速，且仅需最小的电力和实验室设备要求，因此近年来它作为一种可能的解决方案备受关注。然而，由于 LFA 的灵敏度已被证明低于病原体检测的金标准，即细胞培养和实时 PCR，因此 LFA 仍然无法作为有效的 POC 检测方法来预防大流行疫情爆发。提高 LFA 灵敏度的实用方法是在检测步骤之前将目标试剂浓缩在溶液中。在这项研究中，考察了由非离子表面活性剂 Triton X-114 组成的双水相胶束系统，用于在 LFA 之前浓缩模型病毒，即噬菌体 M13（M13）。两种共存胶束相的体积比被操纵以将 M13 浓缩在上部、胶束贫相中。浓缩步骤使该检测方法的 M13 检测限有效提高了十倍，从 5×10(8)噬菌斑形成单位（pfu）/mL 提高到 5×10(7)pfu/mL。未来，可以进一步操纵体积比以获得目标病毒的更大浓度，从而进一步降低 LFA 的检测限。
