Construction method and application of a novel animal model of hepatitis A virus
A hepatitis A virus and animal model technology, applied in the field of medical biology, can solve the problems of high cost of large animals, scarce primate sources, and high rigid conditions.
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[0048] 1. Preparation of experimental animals: breeding of marmots aged 1 to 3;
[0049] 2. Preparation and detection of the virus: Take woodchuck feces and add PBS to prepare a 1:10 (W / V) feces suspension, add chloroform with 1 / 10 of the volume of PBS, and centrifuge at 8000rpm after shaking to get the supernatant to remove solids Impurities, the supernatant was passed through a 0.22 μm filter to remove particulate impurities as much as possible, and the stool suspension was divided and stored in a -80°C refrigerator for later use.
[0050] Adopt upstream primer SEQ ID NO:9 after above-mentioned stool suspension is extracted nucleic acid; Downstream primer SEQ ID NO:10 carries out PCR amplification, and amplified product is carried out sequencing, is determined to be novel hepatitis A virus in the present invention; Primer sequence is as follows Table 3 shows:
[0051] table 3:
[0052] Primer
[0053] 3. Quantification of virus: after the nucleic acid is extracted ...
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