Molecular marker discovery and detection for blinding eye disease.

Accurate, sensitive, and specific detection of molecular markers in intraocular fluid will facilitate the early discovery, diagnosis, and intervention of eye diseases. In this study, a total of 168 participants were recruited and divided into two distinct cohorts: discovery and verification. In the discovery phase, proteomic analysis identified MCP-1 in aqueous humor as a potential molecular marker for blinding eye disease. We further developed a molecular detection technology for the marker based on biolayer interference sensing. The technology utilizes a sandwich strategy with one-to-one pairing of two different biorecognition molecules for MCP-1. It also incorporates automation, high throughput, and real-time monitoring, achieving highly selective recognition and accurate analysis of MCP-1. It demonstrates a low detection limit (0.16 pM), good reliability (R2 = 0.995), and a wide analytical range (0.244-1000 pM) for MCP-1 in human aqueous humor samples. Crucially, in the verification phase with 150 subjects, the technology achieved a high detection rate (95.0%) for patients with age-related macular degeneration and high myopia cataract in under 30 min, and was able to further differentiate between them with a specificity of 86.0%. Therefore, the developed molecular detection technology may provide a robust, convenient, and valuable solution for widespread screening, early discovery, and differential diagnosis of blinding eye diseases.