RESUMO
This study introduces a one-pot isothermal amplification assay for ultrasensitive analysis of terminal deoxynucleotidyl transferase (TdT) activity. The system realizes recycled activation of CRISPR/Cas12a, enabling exceptional signal amplification. This approach maximizes the simplicity of the detection method, offering a promising avenue for molecular disease diagnosis.
Assuntos
Sistemas CRISPR-Cas , DNA Nucleotidilexotransferase , Técnicas de Amplificação de Ácido Nucleico , DNA Nucleotidilexotransferase/metabolismo , Sistemas CRISPR-Cas/genética , HumanosRESUMO
We integrate recombinase polymerase amplification (RPA) with CRISPR/Cas9-initiated nicking rolling circle amplification (CRISPR/Cas9-nRCA) for detecting Staphylococcus aureus. This approach utilizes a unique dimeric G-triplex structure, demonstrating firstly enhanced ThT fluorescence for target detection. The proof-of-concept study introduces a new avenue for integrating isothermal amplifications with CRISPR/Cas9 in the fields of pathogen detection and disease diagnosis.
RESUMO
This study introduces an allosteric palindromic hairpin probe (APHP)-based dual-mode interactive strand displacement amplification (DMI-SDA) system for ultrasensitive detection of microRNA-155. The system achieves exceptional signal amplification and improved signal preservation using dimeric G-triplexes as signal reporters, enabling robust detection of miRNA-155, representing a promising avenue in molecular diagnosis.