Array Detection of Multiple Pathogenic Microorganisms Based on Catalytic Hairpin Assembly
This study establishes a new method for the simultaneous detection of multiple microorganisms using a catalytic hairpin assembly reaction array.The method employs a dual-target strategy.The first target is a conserved sequence of prokaryotic 16S rRNA,used for the preliminary identification of prokaryotes.The second targets are specific sequences for Escherichia coli,Listeria monocytogenes,and Staphylococcus aureus,used for confirming the microbial species.Upon dual-target identification,the trigger chain is released to initiate the catalytic hairpin assembly reaction,cyclically opening hairpin probes labeled with FAM fluorescent groups and quenching groups,thereby generating a fluorescent signal.After optimizing reaction conditions and evaluating signal stability,a working curve of the target sequence detection was established.The method achieves the identification and quantitative detection of multiple microorganisms through the fluorescent signal and establishes a detection array.It demonstrates a good linear relationship within the target concentration range of 3-50 nmol/L and a detection limit of 0.001 nmol/L.In array detection,this method effectively distinguishes three target microorganisms,providing a new technical approach for the simultaneous detection of multiple pathogenic microorganisms.
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