Tetrahedral framework nucleic acids linked CRISPR/Cas13a signal amplification system for rare tumor cell detection.

The sensitive and accurate detection of rare tumor cells provides precise diagnosis and dynamic assessment information in various tumor spectrums. However, rare tumor cells assay is still a challenge due to the exceedingly rare presence in the blood. In this research, we develop a fluorescent approach for the identification of rare tumor cells based on a combination of immunosorbent capture and a three-step signal amplification strategy. First, rare tumor cells are captured by immunoadsorption on 96-well plates. Second, self-synthesized tetrahedral framework nucleic acids (tFNAs) spontaneously anchor into the lipid bilayer of rare tumor cells, resulting in a "one to more" amplification effect. Then, the double-stranded DNA (dsDNA) binds to the vertices of the tFNAs and generates a large amount of target RNA by T7 polymerase, which is the secondary signal amplification. Finally, the target RNA activates the collateral cleavage ability of CRISPR/Cas13a, and the reporter RNA is cleaved for third signal amplification. The detection limit of the proposed method is down to 1 cell mL-1. Furthermore, the tFNAs-Cas13a system is also shown to be capable of detecting rare tumor cells in spiked-in samples and clinical blood samples. This platform enables speedy detection of rare tumor cells with high sensitivity and good specificity, and shows great potential for tumor diagnosis.

Rapid and high-throughput testing of antifungal susceptibility using an AIEgen-based analytical system.

The increasing resistance among fungi to antimicrobials are posing global threats to health. Early treatment with appropriate antifungal drugs guided by the antifungal susceptibility testing (AFST) can dramatically reduce the mortality of severe fungal infections. However, the long test time (24-48 h) of the standard AFSTs cannot provide timely results due to the slow growth of the pathogen. Herein, we report a new AFST that is independent of growth rate analysis using a luminogen with aggregation-induced emission characteristics (AIEgen) named DMASP. DMASP is a water-soluble small-molecule probe that can readily penetrate the dense fungal cell wall. Based on its mitochondria-targeting ability and AIE characteristics, fungal activity can be dynamically indicated via real-time fluorescence monitoring. This allows fungal susceptibility to various antimicrobials to be assessed within 12 h in a wash-free, one-step manner. This method may serve as a promising tool to rapidly detect possible drug-resistant fungal strain and guide the precise use of antimicrobial against fungal diseases.