▎ 摘　　要

NOVELTY - Method (A) for performing differential diagnosis of African swine fever wild strain and vaccine strain using clustered regularly interspaced short palindromic repeats (CRISPR) technology combined with graphene field effect transistor (gFET) involves (a) preparing gFETs, (b) modifying and immobilizing a reporter probe on the gFET to obtain a CRISPR-binding gFET detection system, (c) extracting pET-32a-ASFV-P72 plasmid and diluting it into multiple different concentration gradients, and (d) adding a DNA to-be-detected, CRISPR RNA (crRNA), CRISPR associated 12a (Cas12a) and its buffer to the CRISPR-binding gFET detection system, incubating at 37-37.5°C, observing displacement of Dirac point, and establishing the corresponding relationship between the degree of back-shift of the Dirac point and the concentration of the template DNA. USE - The method (A) is useful for performing differential diagnosis of African swine fever wild strain and vaccine strain using CRISPR technology combined with gFET. ADVANTAGE - The method ensures minimum detection concentration of 0.5aM. DETAILED DESCRIPTION - Method (A) for performing differential diagnosis of African swine fever wild strain and vaccine strain using clustered regularly interspaced short palindromic repeats (CRISPR) technology combined with graphene field effect transistor (gFET) involves (a) preparing gFETs, (b) modifying and immobilizing a reporter probe on the gFET to obtain a CRISPR-binding gFET detection system, (c) extracting pET-32a-ASFV-P72 plasmid and diluting it into multiple different concentration gradients, and (d) adding a DNA to-be-detected, CRISPR RNA (crRNA), CRISPR associated 12a (Cas12a) and its buffer to the CRISPR-binding gFET detection system, incubating at 37-37.5°C, observing displacement of Dirac point, and establishing the corresponding relationship between the degree of back-shift of the Dirac point and the concentration of the template DNA. The method involves using reaction system comprising 5-8 μl crRNA, 5-8 μl buffer, 36-40 μl RNase-free water, 1-3 μl Cas12a and 5-10 μl template DNA. An INDEPENDENT CLAIM is included for a method (B) for performing differential diagnosis of strong and weak strains of African swine fever using CRISPR technology in combination with gFET, which involves preparing gFETs, modifying and immobilizing the reporter probe on the gFET to obtain a CRISPR-binding gFET detection system, extracting the pET-32a-ASFV-P72/MGF/CD2V plasmid and keeping it quantitatively at the same concentration, using different kinds of template DNA, crRNA, Cas12a and its buffer to form a reaction system, adding each of the reaction systems to the CRISPR-combined gFET detection system, incubating at 37°C, and observing for the shift of the Dirac point.