SARS-CoV Intranasal Mouse Model

A dose of 10⁵ 50% TCID₅₀ of SARS-CoV (Urbani) was administered intranasally to a lightly anesthetized, 6-wk-old female BALB/c mouse in a total volume of 50 μL [30 (link)]. Two days after inoculation, the mouse was euthanized, and its lungs were removed and homogenized with an EX-Gen Omni GLH homogenizer (Omni International, http://www.omni-inc.com) as a 10% w/v suspension in Leibovitz's L15 Medium (Invitrogen, http://www.invitrogen.com), supplemented with the following antibiotics: 0.4 mg/L piperacillin (Sigma, http://www.sigmaaldrich.com), 0.1 mg/L gentamicin (Invitrogen), and 5 mg/L amphotericin B (Quality Biological, http://www.qualitybiological.com). The lung homogenate was clarified by low-speed centrifugation at 2,000 rpm (650g) for 5 min, and the supernatant was administered intranasally to three naïve mice. The process of intranasal inoculation of three female BALB/c mice with pooled, clarified supernatants of 10% lung homogenates collected 2 to 3 d.p.i. was repeated 14 times.