PMC:3291650 / 33398-34365 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3291650","sourcedb":"PMC","sourceid":"3291650","source_url":"http://www.ncbi.nlm.nih.gov/pmc/3291650","text":"Viral infection and determination of viral titers\nMouse adapted IAV X-47 (H3N2; PR8×A/Victoria/3/75) was propagated in MDCK cells. For viral inoculation, mice were anesthetized by i.p. injection with ketamine (12 mg/kg) and xylazine (60 mg/kg) and 50 µl X-47 diluted in PBS was administered intranasally. For lethal and sublethal infection, mice received respectively 2-LD50 or 0.05-LD50 X-47. To determine pulmonary viral titers, median tissue culture infectious dose (TCID50) was measured as follows: lungs were homogenized with a Polytron homogenizer (Kinematica) in PBS. Eight-fold serial dilutions of lung homogenates were incubated on MDCK cells for 5 days in DMEM supplemented with trypsin (1 µg/ml), 2 mM L-glutamine, 0.4 mM sodium pyruvate and antibiotics. 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