Abstract
In traditional colorimetric lateral flow immunoassay (LFI) using gold nanoparticles (AuNPs) as a probe, additional optical transducers are required to quantify the signal intensity of the test line because it presents as a single red-colored line. In order to eliminate external equipment, the LFI signal should be quantifiable by the naked eye without the involvement of optical instruments. Given this objective, the single line test zone of conventional LFI was converted to several spots that formed herringbone patterns. When the sandwich immunoassay was performed on a newly developed semi-quantitative (SQ)-LFI system using AuNPs as an optical probe, the spots were colorized and the number of colored spots increased proportionally with the analyte concentration. By counting the number of colored spots, the analyte concentration can be easily estimated with the naked eye. To demonstrate the applicability of the SQ-LFI system in practical immunoanalysis, microalbumin, which is a diagnostic marker for renal failure, was analyzed using microalbumin-spiked artificial urine samples. Using the SQ-LFI system, the calibration results for artificial urine-based microalbumin were studied, ranging from 0 to 500 μg/mL, covering the required clinical detection range, and the limit of detection (LOD) value was calculated to be 15.5 μg/mL. Thus, the SQ-LFI system provides an avenue for the realization of an efficient quantification diagnostic device in resource-limited conditions.
Highlights
The development of an accurate and user-friendly diagnostic device is one of the most significant objectives in the clinical field
We demonstrated that our SQ-lateral flow immunoassay (LFI) system allows for instrument-free immunoanalysis that can be applied to practical diagnoses and efficiently manufactured at conventional facilities
We introduced the concept of a box-stacking animation to the newly developed semi-quantitative LFI (SQ-LFI)
Summary
The development of an accurate and user-friendly diagnostic device is one of the most significant objectives in the clinical field. In the academic and industrial sectors, extensive research has proceeded to develop efficient diagnostic tools, such as point-of-care testing (POCT) biosensors [1,2,3,4,5,6,7]. To develop an efficient POCT-type immunosensor, researchers have focused on the lateral flow immunoassay (LFI) method, which is an immunochromatographic test using a nitrocellulose (NC) membrane as a substrate [8,9,10,11,12,13,14,15]. Despite the merits of LFI, traditional colorimetric AuNP-based LFI is still used in limited tests that only require simple negative or positive results, such as pregnancy and sexually transmitted Since the operation of LFI is automatically accomplished by capillary force and the result can be verified with the naked eye, LFI has merits such as low cost of operation, easy handling, and operation without external instruments.
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