Ternary Card Hypercube Pooling for PCR Testing in Future Pandemics: Tutorial on How to Identify infected patients by efficient pooling strategy (Preprint)

BACKGROUND Sample pooling is an essential strategy for optimizing polymerase chain reaction (PCR) resources during infectious disease outbreaks, especially in the beginning. While high-dimensional hypercube pooling strategies—such as those recently highlighted in Nature—offer superior efficiency in low-prevalence settings, they are difficult to implement in practice. The human cognitive and physical limitation to three-dimensional environments makes manual execution of four- or five-dimensional sample arrays prone to significant operational error. OBJECTIVE To develop and evaluate a novel "Ternary Card Hypercube Pooling" strategy that simplifies the implementation of multidimensional pooling, making it accessible for laboratory personnel without compromising mathematical efficiency. METHODS We integrated logic from ternary card games (based on sets of three attributes) to create a visual and physical framework for hypercube pooling. This method maps high-dimensional coordinates onto a simplified "card" system, allowing laboratory technicians to organize and track samples using intuitive pattern recognition rather than complex multidimensional mapping. RESULTS The Ternary Card method successfully translates the efficiency of hypercube pooling into a user-friendly workflow. It maintains the high performance of traditional hypercubic algorithms—allowing for rapid identification of positive samples in a single step in the majority of cases—while significantly reducing the risk of manual pipetting errors and the need for specialized automated equipment. CONCLUSIONS The Ternary Card Hypercube Pooling strategy bridges the gap between theoretical mathematical efficiency and practical laboratory application. By reducing the complexity of sample handling, this method provides a scalable solution for increasing PCR throughput in response to future pandemics, particularly in resource-limited settings. CLINICALTRIAL NA