Development of a Quantitative Diagnostic Kit based on Fluorescence Immunoassay for Severity Diagnosis in Patients with Inflammatory Dry Eye Disease

Objectives: Dry eye syndrome is a common condition in modern society that causes symptoms such as dryness, foreign body sensation, fatigue, and pain.Initially, symptoms can be improved with simple treatments; however, when accompanied by inflammation, antibiotic therapy is required, necessitating clear tests to distinguish between inflammatory and noninflammatory dry eye syndrome.Currently, clinical assessments such as the Schirmer test and tear break-up time are used for diagnosis, but they have limitations in determining the presence of inflammation.Therefore, this study aimed to develop a quantitative diagnostic kit based on fluorescent immunoassay to diagnose the presence and severity of inflammation.Methods: The developed kit utilizes fluorescent dye as a signaling molecule and employs a sandwich immunoassay method with antibodies fixed on a nitrocellulose membrane to detect Matrix metalloproteinase-9 in tears.Results: As a result, the quantifiable range was found to be 0-100 ng/mL, with a limit of detection of 0.17 ng/mL and a limit of quantification of 0.5 ng/mL.Additionally, an interference test was conducted to verify whether commonly used artificial tears affect the diagnostic system; the results confirmed that no interference from artificial tears was observed. Conclusion:This study establishes a quantitative diagnostic system that overcomes the limitations of existing methods, allowing for clear differentiation of the presence and severity of inflammation in tears, which is expected to contribute to more accurate diagnosis, appropriate treatment, and prevention.