Duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcine circoviruses type 2 and type 3
A technology of porcine circovirus and detection kit, applied in the field of animal virology and molecular biology, to achieve high sensitivity and easy detection method
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[0022] 1 Materials and methods
[0023] 1.1 Viruses and clinical samples
[0024] Porcine parvovirus (PPV), porcine pseudorabies virus (PRV), Escherichia coli, Staphylococcus aureus, Haemophilus para, and Actinobacillus pleuropneumoniae strains or strains are all provided by the laboratory. Clinical samples were collected from the main organs (lymph nodes, spleen, lungs, kidneys, etc.) of dying pigs suspected of being infected with circovirus in pig farms in various places in Guangxi, dead pigs, or pigs that were slaughtered. About 0.5g to 1.0g of tissue samples were cut , placed in a sterilized mortar, fully ground into a homogenate, added sterilized 1mol / LPBS solution to make a 1:5 emulsion suspension, and repeated freezing and thawing 3 times. Centrifuge at 10 000 r / min for 5 min, collect the supernatant, transfer to a 1.5 mL centrifuge tube and number for DNA extraction or store at -20°C for later use.
[0025] 1.2 Main reagents
[0026] pMD18-T Vector, Bam HI, Hin ...
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