METHOD AND KIT FOR DIFFERENTIALLY DETECTING PORCINE PSEUDORABIES VACCINE VIRUS AND WILD VIRUS

Disclosed is a kit for detecting a UL52 gene and/or a gE gene of a porcine pseudorabies virus, including: a forward primer, a middle probe and a reverse primer which are used to amplify a UL52 gene sequence of the porcine pseudorabies virus and/or a forward primer, a middle probe and a reverse primer which are used to amplify a gE gene sequence of the porcine pseudorabies virus by means of recombinase polymerase amplification, the conventional reagents for recombinase polymerase amplification, and a colloidal gold test strip which includes a sampling pad, a control line, a No. 1 detection line and/or No. 2 detection line. Through the cooperation of the control line, the detection lines, and the primer and probe labeling, a UL52 gene and/or a gE gene of a porcine pseudorabies virus in a sample can be detected rapidly, sensitively and specifically by using the kit. - 37- NFO excises THF, and DNA polymerase Principle ofNFO-RPA extends from the excision site and amplifies a template QNFO (Endonuclease V) Forwardprimer 5 Nucleic acid probe y THF VBlockIC3-spacer 5 - 3 5 labeled with FAM A. 3' 5' Perform isothermal amplification on the template at 38C for 20 Reverse primer min to obtain amplified products labeled with Biotin or Dig Amplified product of a UL52 gene of PRV Amplified product of a gE gene of PRV 8.otrn Principle of colloidal-gold test strip detection Colloidal-gold labeling particles Anti-FAM EFAM jAnti-FAM Amplified product of Amplified product of agE gene of PRV aUL52 gene of PRV Colloidal-gold labeling particles H Anti FAM Dig V n Anti. Anti FAM Y Anti-Dig Boin-Ligand Control line of test strip detection No. 2 detection line No.1 detection line * FAMlabeling 'K Coating anti-antibody(Anti-Anti-FAM) * Biotin labeling +( Coating digoxin antibody(Anti-Dig) FAM antibody labeled *Dig labeling ( Coating biotin binding ligand(Biotin-Ligand) by colloidal-gold Fig. 2 - 43-