The HGMS-3D chip and LNA-ddPCR platform for noninvasive detection of KCNJ5 mutations in plasma sEVs demonstrated 64.58% sensitivity, 96.88% specificity, and AUCs of 0.767-0.852 in PA patients.
Observational (n=106)
Does an integrated HGMS-3D chip and LNA-ddPCR platform accurately detect KCNJ5 mutations in plasma sEVs for noninvasive subtyping of primary aldosteronism?
A novel HGMS-3D chip and LNA-ddPCR platform enables noninvasive, highly specific detection of KCNJ5 mutations in plasma sEVs, offering a promising liquid biopsy approach for subtyping primary aldosteronism.
Effect estimate: AUC 0.767-0.852
Primary aldosteronism (PA), a leading cause of secondary hypertension, remains vastly underdiagnosed due to unreliable screening tools. We developed an integrated platform combining a high-gradient magnetic separation three-dimensional (HGMS-3D) chip and locked nucleic acid (LNA)–enhanced droplet digital polymerase chain reaction (ddPCR) for noninvasive detection of potassium inwardly rectifying channel subfamily J member 5 ( KCNJ5 ) mutations in plasma small extracellular vesicles (sEVs). The HGMS-3D chip uses a nickel mesh–based stereoscopic immunoaffinity capture system, achieving a vesicle isolation efficiency 4.4-fold higher than ultracentrifugation. Coupled with LNA-ddPCR, the platform detects KCNJ5 hotspot mutations p.Gly151Arg, (G151R); p.Leu168Arg (L168R) at minor allele frequencies of ≤0.05% ( R 2 ≥ 0.99), overcoming plasma-derived noise. Clinical validation in 106 patients with PA demonstrated 64.58% sensitivity and 96.88% specificity for sEV-based mutation profiling. The assay identified one aldosterone-producing adenoma (APA) case missed by tissue genotyping, achieving area under the receiver operating characteristic curve (AUC) values of 0.767 ~ 0.852 across mutations. This noninvasive approach could enable curative treatment for millions with undiagnosed PA, advancing precision management of endocrine hypertension through sEV-based liquid biopsy.
Wang et al. (Fri,) conducted a observational in Primary aldosteronism (n=106). HGMS-3D chip and LNA-ddPCR platform vs. Tissue genotyping was evaluated on Detection of KCNJ5 mutations (AUC 0.767-0.852). The HGMS-3D chip and LNA-ddPCR platform for noninvasive detection of KCNJ5 mutations in plasma sEVs demonstrated 64.58% sensitivity, 96.88% specificity, and AUCs of 0.767-0.852 in PA patients.