A Fully Integrated ICP-Enriched and Nanozyme-Catalyzed Lateral Flow Assay for cfDNA Detection in Whole Blood

Rapid and sensitive detection of Epstein-Barr virus cell-free DNA (EBV cfDNA) is crucial for early diagnosis and monitoring of nasopharyngeal carcinoma (NPC), but accessibility to screening is limited by complicated and costly conventional DNA isolation and purification approaches. Here, a fully integrated ion concentration polarization (ICP)-enriched and nanozyme-catalyzed lateral flow assay (ICP-cLFA) is developed, enabling total analysis of EBV cfDNA in whole blood samples, with DNA isolation, pre-concentration, and amplification performed on a microfluidic chip, consequently providing the signal readout within 75 min. Specifically, ICP preconcentration and amplification steps, together with target recognition catalyzed by a platinum-decorated mesoporous gold nanosphere (MGNS@Pt) nanozyme, result in an ultralow detection limit of 4 aM in standard cfDNA samples and 100 aM in whole blood from NPC-bearing rats. The high sensitivity and specificity of the ICP-cLFA suggest strong potential for cfDNA screening in resource-limited clinical and field applications.