Methods of assaying the presence of enzymatically active hydrolases (i.e., hydrolytic enzymes) in a sample or specimen are disclosed. In particular, a method of detecting candidiasis by assaying for the presence of enzymatically active aspartic
protease in a sample is provided. In these methods, a sample or specimen is contacted with a
solid support. The
solid support with which the sample is contacted has a reporter
enzyme (i.e., a
signal generating
enzyme) immobilized thereon. The reporter
enzyme is immobilized on the
solid support in a manner such that it is released from the solid support upon action of the enzymatically active
hydrolase if the enzymatically active
hydrolase is, in fact, present in the sample. The sample after having been contacted with the solid support is combined with an indicator. The indicator is any
chemical species which is susceptible to a detectable change, usually a change in color, upon action of the reporter enzyme. A detectable change in the indicator is an indication that the enzymatically active
hydrolase is present in the sample. Moreover, the presence of an enzymatically active hydrolase in a sample may indicate the presence of a particular
pathogen or
disease state such as, for example, candidiasis. In addition to the methods of assaying for the presence of enzymatically active aspartic
protease and other hydrolytic enzymes, a dry, self-contained test device for assaying for the presence of an enzymatically active hydrolase in a sample is disclosed. In particular, a dry, self-contained test device for testing a sample for the presence of candidiasis by assaying for the presence of enzymatically active aspartic
protease is also disclosed. These test devices combine a reporter enzyme immobilized on a solid support, an indicator, and all other reagents and components necessary to achieve a detectable indication of the presence or absence of the enzymatically active hydrolase whose presence is being detected in the sample, and preferred embodiments contain positive and negative controls as well.