1. INFLUENZA TRANSPORT MEDIUM MEDIA PREPARATION TEAM Miguel Garza, Supervisor Martha Thompson, Viral Isolation Laboratory

2. Ingredients Tryptose Phosphate Broth Tryptose Phosphate Broth Gelatin Gelatin Penicillin 250,000 units per mL Penicillin 250,000 units per mL Streptomycin Sulfate 0.25 grams per mL Streptomycin Sulfate 0.25 grams per mL Distilled Water Distilled Water

3. Preparation Combine in a 4 liter flask Combine in a 4 liter flask Dissolve by stirring Dissolve by stirring Autoclave 30 minutes at 121° C Autoclave 30 minutes at 121° C Add aseptically Penicillin 250,000 units and Streptomycin Sulfate 0.25 grams Add aseptically Penicillin 250,000 units and Streptomycin Sulfate 0.25 grams Dispense aseptically 3.0 mls to 1998(aprox.) sterile tubes Dispense aseptically 3.0 mls to 1998(aprox.) sterile tubes

4. Label each tube: INFLUENZA TRANSPORT, 93515, lot number, expiration and Texas Dept. of State Health Services, Austin, TX 78756 Label each tube: INFLUENZA TRANSPORT, 93515, lot number, expiration and Texas Dept. of State Health Services, Austin, TX 78756 Expiration date is 12 months from date of preparation Expiration date is 12 months from date of preparation Store in Media Production Freezer at -25 °C Store in Media Production Freezer at -25 °C Deliver 7 tubes to Virology Team for quality control Deliver 7 tubes to Virology Team for quality control

5. QC of Transport Media Sterility Sterility Toxicity Toxicity Growth assay Growth assay Storage at -70° C for 72 hrs Storage at -70° C for 72 hrs Storage at 4° C for 24 hrs Storage at 4° C for 24 hrs Undilute, 1:5, 1:10 Undilute, 1:5, 1:10 Parallel testing old/new lots Parallel testing old/new lots

6. Viral Transport Media Viral transport medium Viral transport medium Hanks balanced salt solution Hanks balanced salt solution Tryptose-phosphate broth Tryptose-phosphate broth Sucrose-phosphate broth Sucrose-phosphate broth Cell culture medium Cell culture medium Veal infusion broth Veal infusion broth Supplement with a protein or stabilizer, bovine serum albumin or gelatin at 0.5 to 1%. Supplement with a protein or stabilizer, bovine serum albumin or gelatin at 0.5 to 1%. Antibiotics Antibiotics