Detection of mycoplasma contamination.

CFR · 2024-01-01 · US · Title 9

The heart infusion test, using heart infusion broth and heart infusion agar, provided in this section shall be conducted when a test for mycoplasma contamination is prescribed in an applicable Standard Requirement or in the filed Outline of Production for the product. (a) Media additives provided in this paragraph shall be prepared as follows: (1) DPN-Cysteine Solution: (i) Use Nicotinamide adenine dinucleotide (oxidized) and L-Cysteine hydrochloride. (ii) Prepare 1 gram/100 milliliters (ml) purified water (1 percent solution) of each. Mix the solutions together; the (2) Inactivated horse serum—horse serum which has been inactivated at 56 °C for 30 minutes. (b) Heart infusion broth shall be prepared as provided in this paragraph. (1) Dissolve in 970 ml of purified water, 25 grams of heart infusion broth, 10 grams of proteose peptone No. 3, and either 5 grams of yeast autolysate or 5 ml of fresh yeast extract. (2) Add the following: (3) Adjust pH to 7.9 with NaOH, filter sterilize, and dispense 100 ml aliquots into 125 ml flasks and store until needed. (4) Add 2 ml of DPN-Cysteine solution to each 100 ml of broth on day of use. (c) Heart Infusion Agar shall be prepared as provided in this paragraph. (1) Dissolve in 900 ml of purified water by boiling the following: (2) Cool the medium and adjust pH to 7.9 with NaOH. (3) Autoclave the medium. (4) Cool the medium 30 minutes in a 56 °C waterbath. (5) Dissolve 5 grams of yeast autolysate in 100 ml of distilled water, filter steril

official document · source cfr-2024 · id 9:§ 113.28