DNA-Based Loop-Amplifier Fluorescence Signaling Networks Enable Rapid Quantitative Pathogen Detection of...

Periprosthetic microbial bone infection (PMBI) is a serious pathological condition that may lead to terrible consequences without timely therapeutic intervention. However, the clinical management of PMBI is challenged by the difficulties in the efficient and accurate identification of microbial pathogens. For this purpose, we report a DNA-based loop-amplifier fluorescence signaling network (DLFSN) for the efficient and accurate detection of pathogenic microbes in the context of PMBI. The DLFSN system comprises an Amplifier Unit A and a Quantifier Unit B. Specifically, Amplifier Unit A transforms pathogenic microbial genetic materials into fluorescent DNA molecules through an entropy-driven automated reaction loop, leading to efficient and precise amplification of pathogen-specific genomic input signals. Subsequently, Quantifier Unit B converts the quenched fluorescent intermediate products into an activated state, thus allowing the precise identification of the genetic signature of pathogenic microorganisms in a quantitative manner. DLFSN demonstrated high diagnostic accuracy comparable to gold standard PMBI diagnostic technologies, including tissue section staining and qPCR, while the actual assay time was much lower at only around 30 min for real-life patients. The DLFSN system provides a promising precision diagnostic tool for early PMBI diagnosis, holding promise to improve orthopedic outcomes in the clinic.