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Alt Text Acknowledgement

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Final Abstract

Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique known for its specificity and efficiency in less controlled environments than laboratories. Designing novel LAMP reagents requires strict contamination control, and unlike the more commonly automated polymerase chain reaction, LAMP’s heightened sensitivity poses unique challenges to automation. To address this, we optimized automation procedures for LAMP using a robotic pipettor without altering the core assay itself. This project focuses on minimizing contamination using sealed plates, reducing operator intervention by automating pipetting, and preventing enzyme degradation using cooling plates. Our approach employs pre-slit pierceable seals and cooling blocks to prevent cross-contamination between wells and maintain primer integrity, enhancing the system’s scalability for large sample sizes. The implementation of this automated workflow resulted in an 81.6% reduction in processing time, reducing LAMP assay duration from 1 hour for 16 reactions in a 96-well plate to just 15 minutes. Comparative analysis between manual and automated runs shows no significant difference in reaction profiles, and initial evaluations indicate acceptable levels of cross-contamination. By extending this automation workflow, we aim to create a fully autonomous 24-hour LAMP screening system by integrating a nucleic acid synthesizer for on-demand primer production. Furthermore, this system will interface with AI-based experimental design models, enabling autonomous optimization of LAMP reagent design and assay development. By eliminating manual intervention, the diagnostic process is streamlined, accelerating assay development and enhancing response capabilities for emerging pathogens. This marks a significant advancement in biomolecular diagnostics, with potential for continuous innovation to address evolving public health needs.

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