Assays for trichomonal and other hydrolases

A method for detecting Trichomoniasis vaginalis in vaginal fluid, said method comprising:

• (a) wetting an implement capable of retaining liquid with said vaginal fluid at a pH within the range of 2 to 3.5 and applying said wetted implement to a solid-phase conjugate comprising a reagent covalently bonded to a member selected from the group consisting of lysine and arginine residues at the C-terminus of a peptide in a manner resulting in the retention of said reagent by said wetted implement upon cleavage of said reagent from said peptide, said reagent being a species that when released from said binding member and contacted with an indicator causes a detectable change in said indicator;
• (b) applying said implement, after having been applied to said solid-phase conjugate, to a solid-phase form of said indicator;
• and
• (c) detecting whether said detectable change has occurred on said implement, the occurrence of said detectable change being an indication of the presence of Trichomoniasis vaginalis in said vaginal fluid.

A method for detecting Trichomoniasis vaginalis in vaginal fluid, said method comprising:

• (a) drawing a sample of said vaginal fluid;
• (b) extracting from said sample a supernatant that is devoid of particulate matter greater than 20 microns in diameter;
• (c) contacting said supernatant with a substrate that produces a detectable change upon hydrolytic cleavage;
• and
• (d) observing whether said detectable change has occurred, the occurrence of said detectable change being an indication of the presence of Trichomoniasis vaginalis in said vaginal fluid.

A test device for assaying a sample of bodily fluid for the presence of a water-soluble hydrolase, said test device comprising:

• a flow passage having a defined application site for application of said sample;
• a solid porous material, retained within said flow passage, that retains particulate matter greater than 20 microns in diameter while allowing aqueous liquids to pass;
• an indicator deposited in solid form in said flow passage, said indicator being soluble in an aqueous liquid and producing a detectable change when contacted with an activated reagent;
• and
• a conjugate deposited in solid form in said flow passage, said conjugate comprising said reagent covalently bonded to a binding member, said reagent releasable from said binding member by enzymatic hydrolytic cleavage action and thereby activated;
• at least one of said indicator and said conjugate being spatially separated from said application site by a distance sufficiently great that a detectable change in said indicator is caused only by hydrolytic activity of said hydrolase in a fraction of said sample remaining after said particulate matter has been removed from said sample by said solid porous material.

Test apparatus for assaying a sample of bodily fluid for the presence of a hydrolytic enzyme, said test apparatus comprising:

• (a) an inert solid support having a surface on which test materials are deposited in a plurality of spatially separated regions, said regions comprising:
• (i) a first region having deposited thereon a conjugate comprising a reagent covalently bonded to a binding member and activatable by hydrolytic cleavage from said binding member, and
• (ii) a second region having deposited thereon an indicator that undergoes a detectable change when contacted with said reagent in activated form, and
• (b) an absorptive implement that is wettable by bodily fluid and, when so wetted and rubbed against said first and second regions, causes test materials residing in said first and second regions to adhere to said implement.