Porcine epidemic diarrhea virus, and culture method and application thereof
A technology for porcine epidemic diarrhea and viruses, applied in the direction of antiviral agents, viruses/bacteriophages, and microbial-based methods, which can solve the problems of limited selection and lack of broad immunogenicity
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Embodiment 1
[0030] Embodiment 1 GDSJ / 2012 virus strain obtains
[0031] 1. Collection and processing of disease materials
[0032] Seven fresh feces of piglets with suspected viral diarrhea in a pig farm in Guangdong Province were collected, diluted with PBS (phosphate buffer saline) at a weight-to-volume ratio of 1:10 after numbering, shaken and mixed evenly, and repeatedly frozen and thawed 3 times , centrifuged at 8000r / min (rotation per minute) for 10 min (minutes), took the supernatant, sterilized it with a filter with a pore size of 0.22 μm (microns), and stored the resulting treated solution in a -70°C refrigerator for later use.
[0033] 2. RT-PCR (reverse transcription polymerase chain amplification reaction) detection of disease materials
[0034] Take 200 μL (microliter) of each of the above treatment solutions numbered 1-7, and use the AxyPrep Body Fluid Virus DNA / RNA Mini Kit produced by Shanghai Baisai Biotechnology Co., Ltd. to extract RNA. The extracted RNA was reverse-t...
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