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Anal Bioanal Chem. 2010 Dec;398(7-8):2955-61. doi: 10.1007/s00216-010-4213-7. Epub 2010 Sep 24.

Enhancing the lateral-flow immunoassay for viral detection using an aqueous two-phase micellar system.

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1
Department of Bioengineering, University of California, 5121 Engineering V, 420 Westwood Plaza, Los Angeles, CA 90095-1600, USA.

Abstract

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.

PMID:
20865404
PMCID:
PMC2990019
DOI:
10.1007/s00216-010-4213-7
[Indexed for MEDLINE]
Free PMC Article
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