CRYOPRESERVATION OF VIRUS ISOLATES (Appendix XII)

1. Record in the computer Freezer Program details of the isolate stored.

a. Select "A. Clinical Records".

b. "Add new entries?": enter "Yes".

c. Enter lab number, ensure patient data is correct, enter date frozen, enter study (ie "VIR"), ensure specimen type is correct.

d. Enter name of isolate to be frozen once.

e. Note the freezer locations in which the frozen isolates are to be placed.

- Freezer "G" & "J" refer to Liquid Nitrogen freezers.

- Shelf no. refers to the canister number.

- Box no. refers to the cane number. Each cane holds 5 freezer vials.

 

2. Discard maintenance medium on monolayer to be frozen.

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3. Add 1.0 mL viral freezing medium (see Appendix VII) to positive cells. Scrape monolayer and mix well with freezing medium.

 

4. Distribute into freezing vial labelled with LIS label.

 

5. Store the vial immediately in liquid nitrogen freezer in the location specified by the freezer program.

 

FREEZING CAP SPECIMENS AND ISOLATES:

1. After planting CAP specimens, add 1 mL of viral freezing medium to the sample and transfer to a freezing vial. Enter into the freezer program as described above using "VQC" for study. Enter "CAPSAM" as isolate #1.

 

2. Freeze one vial of CAP viral isolates after identification using steps 2 - 5 described above. The frozen isolate will be isolate #2 and is entered using the code for the name of the virus. (The original sample and frozen isolate may be on different canes, this is acceptable).

 

Reference

Laboratory Diagnosis of Infectious Diseases. 1988 Volume II. Pg. 268.

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