Abstract
The development of quantitative lateral flow immunoassay test strips involves a lot of research from kit manufacturers’ standpoint. Kit providers need to evaluate multiple parameters, including the location of test regions, sample flow speed, required sample volumes, reaction stability time, etc. A practical visualization tool assisting manufacturers in this process is very much required for the design of more sensitive and reliable quantitative LFIA test strips. In this paper, we present an image-based quantitative evaluation tool determining the practical functionality of fluorescence-labelled LFIA test cartridges. Image processing-based algorithms developed and presented in this paper provide a practical analysis of sample flow rates, reaction stability times of samples under test, and detect any abnormalities in test strips. Evaluation of the algorithm is done with Glycated Hemoglobin (HbA1C) and Vitamin D test cartridges. Practical sample flow progress for HbA1C test cartridges is demonstrated. The reaction stability time of HbA1C test samples is measured to be 12 min, while that of Vitamin D test samples is 24 min. Experimental evaluation of the abnormality detection algorithm is carried out, and sample flow abnormalities are detected with 100% accuracy while membrane irregularities are detected with 96% accuracy.
Highlights
IntroductionLateral flow immunoassays (LFIA) form the most widely used point-of-care (PoC) diagnostic tools
The developed algorithm, along with the mentioned hardware setup, forms a portable, simple-to-use evaluation tool, facilitating the research involved in the design and manufacturing of fluorescence-labelled quantitative lateral flow immunoassay test cartridges
The tool efficiently detects the presence of any abnormalities, which could be flow-related or irregularities in the manufactured test membranes
Summary
Lateral flow immunoassays (LFIA) form the most widely used point-of-care (PoC) diagnostic tools. The prefabricated LFIA test strips can be employed for qualitative as well as quantitative study of the target analytes [1]. Infectious diseases like Malaria, HIV, Dengue, and non-communicable diseases, including Cardiac issues and Diabetes, etc., can be detected with these portable, one-time-use devices. LFIA technology can help in providing a quantitative estimate of haptens, proteins, vitamin and hormone levels, etc., in serum/blood samples [1,2]. Traditional diagnostic procedures like high-performance liquid chromatography (HPLC), mass spectroscopy, etc., demand expensive laboratory setups and are time-consuming. These devices involve extensive cleaning procedures and require skilled professionals for performing the tests [3].
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