| 摘要 |
Methods of assaying for 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 i n a sample is provided. In these methods, a sample or specimen is contacted with a soli d support. The solid support with which the sample is co ntacted has a reporter enzyme (i.e., a signal generating enzyme) immobilized thereon . The reporter enzyme is immobilized the solid support in a manner such that it is released from the solid support upon action of t he enzymatically active hydrolase if the enzymatically a ctive hydrolase is, in fact, present in the sample. The sample after having been c ontacted with the solid support is combined with an indi cator. The indicator is any chemical species which is susceptible to a detectable c hange, usually a change in color, upon action of the rep orter enzyme. A detectable change in the indicator is an indication that the enzym atically active hydrolase is present in the sample. More over, the presence of an enzymatically active hydrolase in a sample may indicate t he presence of a particular pathogen or disease state su ch as, for example, candidiasis. In addition to the methods of assaying for the presenc e of enzymatically active aspartic protease and other hy drolytic enzymes, a dry, self-contained test device for assaying for the presence of an enzymatically active hydrolase in a sample is disclos ed. In particular, a dry, self-contained test device for testing a sample for the p resence of candidiasis by assaying for the presence of e nzymatically active aspartic protease is also disclosed. These test devices combine a rep orter enzyme immobilized on a solid support, an indicato r, and all other reagents and components necessary to achieve a detectable indi cation of the presence or absence of the enzymatically a ctive hydrolase whose presence is being detected in the sample, and preferred embo diments contain positive and negative controls as well.
|
