Phase Transition of Wax Enabling CRISPR Diagnostics for Automatic At‐Home Testing of Multiple Sexually Transmitted Infection Pathogens

Abstract Sexually transmitted infections (STIs) significantly impact women's reproductive health. Rapid, sensitive, and affordable detection of these pathogens is essential, especially for home‐based self‐testing, which is crucial for individuals who prioritize privacy or live in areas with limited access to healthcare services. Herein, an automated diagnostic system called Wax‐CRISPR has been designed specifically for at‐home testing of multiple STIs. This system employs a unique strategy by using the solid‐to‐liquid phase transition of wax to sequentially isolate and mix recombinase polymerase amplification (RPA) and CRISPR assays in a microfluidic chip. By incorporating a home‐built controlling system, Wax‐CRISPR achieves true one‐pot multiplexed detection. The system can simultaneously detect six common critical gynecological pathogens (CT, MG, UU, NG, HPV 16, and HPV 18) within 30 min, with a detection limit reaching 10 ⁻¹⁸ M. Clinical evaluation demonstrates that the system achieves a sensitivity of 96.8% and a specificity of 97.3% across 100 clinical samples. Importantly, eight randomly recruited untrained operators performe a double‐blinded test and successfully identified the STI targets in 33 clinical samples. This wax‐transition‐based one‐pot CRISPR assay offers advantages such as low‐cost, high‐stability, and user‐friendliness, making it a useful platform for at‐home or field‐based testing of multiple pathogen infections.