Abstract: A biochemical test system is disclosed for use in a biological fermentation sterilization indicator. The carbohydrate component of a nutrient broth media is removed from the media. The carbohydrate component may be disposed in the cavity of an encapsulated sealed glass ampul which contains a non-carbohydrated test solution. Microorganism spores are also disposed in the cavity. Upon completion of sterilization the ampul is broken; the contents mix. Fermentation of viable spores yields growth and color change and is a positive test (unsatisfactory sterilization); no fermentation yields no growth, no color change and satisfactory sterilization. The separation of carbohydrate prevents non-microorganism fermentation in the test reaction, and the appearance of a "false positive".