The invention relates to the technical field of
virus detection, and discloses a crRNA primer pair for PPV detection, a CRISPRCas12a
system and an application method.The method comprises the following steps that S1,
tissue sample treatment is conducted, specifically, about 1 g of tissue samples are weighed, 1 mL of PBS is added, the mixture is put into a
grinding bowl to be ground into homogenate, the homogenate is transferred into a sterile centrifugal tube, and
centrifugation is conducted for 3 min in a centrifugal
machine at the speed of 12000 rpm / min; s2, extraction of
viral nucleic acid: carrying out
nucleic acid extraction on the
virus by using a viral
genome DNA /
RNA extraction kit of a certain biochemical science and technology Co., Ltd; s3, primer design: retrieving a
porcine parvovirus whole
genome (
GenBank ID: NC001718.1) in
GenBank, determining a highly conserved fragment in the
porcine parvovirus whole
genome as a VP2
gene through analysis and comparison by Meglign
software, designing a full-length primer of the VP2
gene by using Primer Premier 5
software, and designing a full-length primer of the VP2
gene according to the highly conserved fragment VP2 gene in the
porcine parvovirus whole genome; the crRNA primer of the porcine
parvovirus VP2 gene is designed by using Cpf1 primer
design software in a
CRISPR-DT website, and is entrusted to be synthesized and produced by a biological company.