Piezoelectric printing-based droplet digital PCR for noninvasive and sensitive detection of Helicobacter pylori in...

Helicobacter pylori (H. pylori), classified by the WHO as a Group I carcinogen, is a major contributor to progressive gastric diseases, and its growing resistance to antibiotics has severely undermined first-line eradication therapies. Conventional stool-based diagnostics face three key limitations: limited ability to detect resistant strains, labor-intensive culture workflows, and insufficient sensitivity in inhibitor-rich matrices. These shortcomings underscore the urgent need for a noninvasive, highly sensitive, and inhibitor-tolerant platform that enables simultaneous pathogen detection and resistance assessment. To address this unmet need, we developed a piezoelectric printing-driven ddPCR platform. By optimizing piezoelectric parameters and oil-phase surfactant formulations, we achieved uniform and stable droplet generation, producing a high-density microarray of ∼30,000 droplets in 3 min. The droplets exhibited excellent thermal stability, enabling robust digital PCR amplification. Using stool samples, the platform simultaneously detected H. pylori infection and clarithromycin resistance mutations. For H. pylori detection, it exhibited a sensitivity of 1 copy/μL, a quantitative linear range of 10⁰-10⁵ copies/μL (R² = 0.99), and 100 % concordance with commercial assays. This technology provides a scalable, cost-effective, and clinically practical solution for stool-based pathogen detection and antimicrobial resistance surveillance, advancing precision diagnostics in both clinical and public health settings.