Point of care diagnosis of dry eye disease with a sensitive immunoassay for dual biomarker detection

Dry eye disease (DED) is a multifactorial eye disease with few effective methods for clinical diagnosis and treatment. It is the most common eye disease with significant health challenges of the unprecedented aging population. Recent proteomic studies and clinical research have led to the discovery of several biologically relevant biomarkers, with increased levels of Interleukin-6 (IL-6) and decreased levels of lactoferrin being clinically validated in the progression of DED. In this study, a sensitive point of care (POC) DED diagnostic method was developed for targeting dual biomarkers of IL-6 and lactoferrin in the tear samples. A paper-based lateral flow immunoassay (LFIA) was established in a double-antibody sandwich fashion with colloid gold nanoparticles acting as probes. The minimal detection concentrations were 0.1 ng/ml and 10 ng/ml for IL-6 and lactoferrin, respectively. Separated conventional ELISA tests were also performed with data confirming results from the LFIA tests. A trial study was conducted with 20 tear samples from DED patients and healthy controls. All DED tear samples exhibited significantly higher levels of IL-6 and decreased levels of lactoferrin, as compared to the normal controls. A quantitative analysis of LFIA images was carried out using ImageJ software for an accurate data interpretation. This dual biomarker detection method is sensitive and affordable with quick turnaround time. Design of a larger clinical study in the future can further validate this POC assay for early diagnosis as well as patients' self-management of chronic states of DED.