Methods in molecular biology (Clifton, N.J.)

Fast and Accurate Detection of Phytoplasma Diseases Using a CRISPR-Based Test with Constant-Temperature DNA Amplification

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Abstract

The DETECTR assay enables rapid detection of phytoplasma diseases using CRISPR/Cas12a technology combined with isothermal amplification.

  • The assay amplifies target DNA from phytoplasmas using Recombinase Polymerase Amplification (RPA).
  • Cas12a nuclease is activated to cleave both the amplified DNA and fluorescent oligonucleotide reporters after initial target cleavage.
  • This process generates a fluorescent signal that indicates the presence of phytoplasma DNA.
  • Guidelines are included for sample preparation, assay setup, and interpreting fluorescence data.

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