Enhancing the lateral-flow immunoassay for viral detection using an aqueous two-phase micellar system.
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.
Mashayekhi
Foad
F
Department of Bioengineering, University of California, 5121 Engineering V, 420 Westwood Plaza, Los Angeles, CA 90095-1600, USA.
Chiu
Ricky Y T
RY
Le
Alexander M
AM
Chao
Felix C
FC
Wu
Benjamin M
BM
Kamei
Daniel T
DT
D017104:Bacteriophage M13 / Q000302:isolation & purification*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008823:Micelles; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry*
D008823:Micelles; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40