Light source-free smartphone detection of salivary cortisol via colorimetric lateral flow immunoassay using a photoluminescent film

Biosens Bioelectron. 2025 Mar 1:271:116971. doi: 10.1016/j.bios.2024.116971. Epub 2024 Nov 20.

Abstract

An accurate assay was developed by integrating a novel lateral flow immunoassay (LFIA) design, smartphone-based photoluminescence detection, and computer-aided analysis using machine learning algorithms for the quantitative measurement of cortisol levels in human saliva samples. The unique LFIA strip incorporates a photoluminescent film, which enables photoluminescence detection without an external light source, beneath a nitrocellulose membrane. A smartphone is used to capture images of the LFIA test strips, and specific regions in the captured images are analyzed. The digitized data are then processed using a computer. Machine learning algorithms were employed to interpret the data and quantify cortisol levels in saliva samples obtained from 14 volunteers. The developed assay was shown to be highly accurate, and a low average difference of 18.12% was observed between the predicted cortisol levels and those measured using an established enzyme-linked immunosorbent assay (ELISA) in real saliva samples. The assay has a calculated limit of detection of approximately 139 pg/mL. Furthermore, the strong correlation (r = 0.935) between the results of the developed assay and the ELISA results supports its validity.

Keywords: Competitive lateral flow immunoassay; Cortisol; Human saliva; Photoluminescence detection; Smartphone.

MeSH terms

  • Biosensing Techniques* / instrumentation
  • Colorimetry* / instrumentation
  • Enzyme-Linked Immunosorbent Assay
  • Equipment Design
  • Humans
  • Hydrocortisone* / analysis
  • Immunoassay / instrumentation
  • Immunoassay / methods
  • Limit of Detection
  • Machine Learning
  • Saliva* / chemistry
  • Smartphone*

Substances

  • Hydrocortisone