When performing virus identification or vaccine potency determinations, the virus should first be titrated to determine the dosage unit or vaccine titer.

Operation method

dilution

Move

1. Make serial 10-fold dilutions of the virus (one new pipette for each dilution) in maintenance solution (0.5% Hank's solution with hydrolyzed lactic protein).

2. For titrations of l0-6 to 10-8, arrange 8 tubes in a rack: 5 tubes of 1.8 mL of Maintenance Solution for l0-1 to 10-5, and 3 tubes of 4.5 mL of Maintenance Solution for l0-6 to 10-8. Pipette 0.2 ml of virus stock solution into the first tube, then blow with a new pipette and remove 0.2 ml into the second tube, and then continuously dilute to the fifth tube in the same way. From tube 5, pipette 0.5 ml into tube 6 and blow through a new pipette to remove 0.5 ml into tube 7; at the same time, use the same pipette to remove 1 ml of 10-8 virus solution and inoculate the cell tubes with 4 tubes at each dilution, and so on.

3. Observe for several days after inoculation, depending on how quickly the lesions appear.

4. The dilution at which half of the cell tubes show lesions is 50% of the infected units (called TCID50).

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