While the virus is raging
in pursuit of the potential of our hot springs for healthcare,
influenza inactivation experiments using hot springs
March this year It was held on the 11th.
The test method is as follows.
a) Add 0.01 mL of Ruflox to 0.99 mL of purified water to prepare a 100-fold diluted solution.
In addition, 0.15 mL of Ruflox 100-fold diluted solution was added to 2.85 mL of purified water to create a 2000-fold diluted solution.
b) Dispense 1.08 mL of Ruflox 2000-fold diluted solution into a tube, and mix 0.12 mL of influenza virus solution prepared at 5-10 x 10⁵ pfu/mL using phosphate buffer solution (PBS). , and the stirred one was used as the test solution.
c) The test solution was allowed to stand at room temperature to confirm the reaction.
Collect 0.12 mL of solution from the test solution at predetermined time intervals (immediately after, 1 minute, 5 minutes),
1.08 mL of Dulbecco's modified Eagle medium (FBS-DMEM medium) containing 0, 2% bovine serum mixed with
Dilution with 0.2% FBS-DMEM medium was repeated multiple times to create a 10-fold serial dilution series.
Ten-fold serial dilutions were added dropwise to each 1mL/well of host cells prepared in advance, and infected at 37°C under 5Z% CO₂ for 1 hour.
d) After virus infection, the cell supernatant was replaced with 0.8% oxoid solution, and
cultured at 37°C, 5% CO₂ for 2 days. After visually confirming the formation of plaques,
fix with a 5% glutaraldehyde solution and stain with methylene blue, and
measure the virus infection titer based on the data of the number of plaques formed.
e) This test was conducted twice, and the influenza virus inactivation rate of Ruflox 2,000-fold diluted solution was evaluated based on the average value of the two times.
More than 99.988%virus inactivation was confirmed immediately after the action of the 2000-fold dilution of the
hot spring undiluted solution. .