Issue 18, 2022

Immuno-chromatic probe based lateral flow assay for point-of-care detection of Japanese encephalitis virus NS1 protein biomarker in clinical samples using a smartphone-based approach

Abstract

Lateral flow assays (LFAs) are one of the most economical, point-of-care (PoC) diagnostic assays that exploit the colorimetric properties of gold nanoparticles (AuNPs). Up to the best of our knowledge, no rapid antigen-based LFA exists for Japanese Encephalitis Virus (JEV) detection. Herein, we have reported a novel portable sandwich-type LFA for on-site detection of the non-structural 1 (NS1) secretory protein of JEV. In-house JEV NS1 antibodies (Abs) were generated and labelled with AuNPs as immunoprobes. A glass fibre membrane conjugate pad was soaked with AuNPs–Ab solution, while the JEV NS1 Ab and anti-rabbit IgG 2° Ab were coated as the test and control lines, respectively, on a nitrocellulose (NC) membrane. Different layers of the LFA were fabricated and various parameters were standardised for optimum colour intensity development. JEV negative serum samples spiked with JEV NS1 Ags (linear range – 1 pg ml−1 to 1 μg ml−1) were applied onto the sample pad and the intensity of the red colour developed on the test line increased with increasing concentration of Ag. The visual limit of detection (LOD) determined from the LFA was 10 pg ml−1, which corresponded to the LOD determined by the graphical data obtained from ImageJ software and the Colorimeter smartphone application. Furthermore, the colorimetric based immunosensor showed minimal non-specific detection of other closely related flaviviral NS1 Ags in the spiked serum, provided a rapid result within 10 min, showed storage stability up to a month at 4 °C, successfully detected the JEV NS1 protein in clinically infected pig serum samples, and hence, may be developed into a PoC screening diagnostic kit for JEV.

Graphical abstract: Immuno-chromatic probe based lateral flow assay for point-of-care detection of Japanese encephalitis virus NS1 protein biomarker in clinical samples using a smartphone-based approach

Article information

Article type
Paper
Submitted
18 Jūl. 2022
Accepted
08 Aug. 2022
First published
10 Aug. 2022
This article is Open Access
Creative Commons BY license

Nanoscale Adv., 2022,4, 3966-3977

Immuno-chromatic probe based lateral flow assay for point-of-care detection of Japanese encephalitis virus NS1 protein biomarker in clinical samples using a smartphone-based approach

A. Roberts, D. Prakashan, H. Dhanze, R. K. Gandham, S. Gandhi and G. T. Sharma, Nanoscale Adv., 2022, 4, 3966 DOI: 10.1039/D2NA00463A

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