Granted Patent B2

US 12,351,856 · App. 17/959,215 · Granted Jul 8, 2025

Inventors: Manuel Rozas Andreu (Puerto Varas, CL); Daniel E. Almonacid Coronado (Lafayette, CA)

Assignee: KURA BIOTECH INC.

C12Q1/34G01N33/56916G01N33/948G01N33/9486G01N30/72G01N2333/924

Abstract

Claims (10)

1. A method for detecting opioids, opiates, cannabinoids, or benzodiazepines in a sample, comprising the steps of:

(a) contacting the sample with an enzyme having β-glucuronidase activity, wherein the enzyme comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO:1 and is derived from Brachyspira pilosicoli , wherein the sample comprises glucuronide metabolites of opioids, opiates, cannabinoids, or benzodiazepines;

(b) incubating the sample with the enzyme to produce opioids, opiates, cannabinoids, or benzodiazepines by hydrolysis or glycosidic bond cleavage of the glucuronide metabolites; and

(c) detecting the opioids, opiates, cannabinoids, or benzodiazepines through any suitable technique.

2. The method of claim 1 , wherein the sample is a biological sample.

3. The method of claim 2 , wherein the biological sample is selected from the group consisting of saliva, whole blood, plasma, urine, hair, skin, teeth, soft tissues, meconium and vitreous humor.

4. The method of claim 1 , wherein the step of incubating the sample with the enzyme is performed for a period between 1 and 30 minutes.

5. The method of claim 1 , wherein the step of incubating the sample with the enzyme is performed at a temperature range between 20 and 60 degrees Celsius.

6. The method of claim 1 , wherein the step of incubating the sample with the enzyme is performed at a range of pH between 4.0 and 9.0.

7. The method of claim 1 , wherein the suitable technique for detecting the opioids, opiates, cannabinoids, or benzodiazepines is selected from the group consisting of LC-MS, GC-MS, HPLC-MS, LC-HRMS, HPLC-HRMS, LC-MS-MS, GC-MS-MS and HPLC-MS-MS.

Assignments (2)

Priority Claims (1)

Continuity (3)

References Cited (42)