A streamlined integrated system integrating lysate release, freeze-dried reagents for multiplex polymerase chain reaction, and intelligent analysis for TORCHes pathogen identification.

Introduction: Current Methods: We utilized a self-prepared lysis buffer for fast nucleic acid extraction from diverse sample types. Primers, probes, and reaction components for six TORCHes pathogens were fabricated into freeze-dried microspheres via vacuum freeze-drying, enabling single-tube multicolor melting curve analysis (MMCA). The companion software, TORCHes-MCAv1.0, was used to automate pathogen typing and result reporting. Evaluation metrics encompassed sensitivity, specificity, precision, stability, and anti-interference ability. The system was validated using 210 clinical samples compared to commercial single-target qPCR, with Sanger sequencing resolving any discrepancies. Results: Nucleic acid extraction and release were completed within 1 min. Freeze-dried microspheres demonstrated exceptional repeatability, with Conclusion: We developed a robust, streamlined integrated TORCHes detection system. Its simplicity, stability, and high-throughput nature make it a valuable complementary tool for confirming active infection and differentiating pathogen subtypes in complex clinical scenarios.