Chicken reovirus variation strain HeB18
A reovirus and vaccine technology, applied in the direction of antiviral agents, virus/bacteriophage, virus antigen components, etc., can solve the problem of decreased protection of chicken reovirus vaccine
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Embodiment 1
[0013] Embodiment 1: the isolation process of virus strain
[0014] In February 2018, in a commercial white-feather broiler farm in Hebei, a 23-day-old white-feather broiler suffered from large-area hock joint distortion, swelling, and hockey stick-shaped legs. Pressure leads to increased death rate. The sick chickens were put to death, the hock joints were cut open, and the joint fluid was dipped with a sterile cotton swab. After being sent to the laboratory, the cotton swabs were rinsed with sterile saline, and sterilized by filtration with a 0.22 μm filter. The above-mentioned filtrate was inoculated into 10 pieces of SPF chicken embryos through the chorioallantoic membrane, and the inoculation amount was 0.2ml / piece. After inoculation, they were placed in a biochemical incubator at 37°C for constant temperature incubation. The chicken embryos that died within 24 hours were discarded, observed continuously for 7 days, and the chorioallantoic membranes of dead and undead c...
Embodiment 2
[0017] Embodiment 2: Identification of He18 strain
[0018] 1. PCR identification
[0019] 1. Extraction and reverse transcription of viral RNA
[0020] Add 300 μl of virus chorion grinding supernatant to a 1.5ml centrifuge tube, add 700 μl Trizol Reagent RNA extraction solution, mix well, and place at room temperature for 15 minutes. Add 200 μl of chloroform, shake and mix thoroughly, and let stand at room temperature for 10 minutes. Centrifuge at 12,000 rpm for 15 min at 4°C, and transfer 500 μl of the supernatant to a 1.5 ml centrifuge tube treated with DEPC. Add 500 μl of pre-cooled isopropanol, let stand at -20°C for 30 minutes, centrifuge at 12,000 rpm for 15 minutes, and discard the supernatant. Add 1ml of 75% ethanol, centrifuge at 8000rpm for 5min, and pour off the liquid. Put it upside down on paper to dry, suspend the pellet in 30 μl of RNA-free water, and store it at -80°C for later use. The extracted RNA was reverse-transcribed according to the instruction ma...
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