Abstract: A sterilization challenge specimen holder is used with a test indicator to challenge sterilization on a consistent basis. The holder includes a body having an internal chamber region and a sleeve positionable within the internal chamber. A cap is sealable on the holder and a plug is positioned in the body. The plug has a wall having a spiral formed groove therein in communication with the internal chamber region. An opening in the holder provides for a flow path for communication between the environs and the internal chamber, and another opening(s) provides for a second flow path for communication between the internal chamber and the inner chamber of the sleeve. An indicator tube carries a biological indicator and has openings therein.