Magnetic bead-assisted one-pot RCA-activated CRISPR/Cas12a electrochemiluminescence biosensor for the detection of citrus Huanglongbing pathogen

A linear detection range of 10 fM-1 nM with a limit of detection of 2 fM was achieved for Candidatus Liberibacter asiaticus gene fragments.

• The biosensor integrates rolling circle amplification and CRISPR/Cas12a activities for enhanced detection.
• Target-initiated amplification leads to the degradation of specific DNA, restoring electrochemiluminescence signals.
• High specificity was demonstrated for detecting the ribonucleotide-diphosphate reductase subunit beta gene fragments.
• The system eliminates the need for multiple incubation steps, streamlining the detection process.
• Satisfactory performance was observed in detecting HLB pathogens in citrus leaf samples.

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