CEDIA® Buprenorphine II Assay For In Vitro Diagnostic Use Only Rx Only 10020849 (3 x 17 mL Kit) 10020850 (65 mL Kit) Intended Use Warnings and Precautions The CEDIA® Buprenorphine II Assay is a homogeneous enzyme immunoassay for the DANGER: Powder reagents contain ≤55% w/w Bovine Serum Albumin (BSA) fragments and qualitative and/or semi-quantitative determination for the presence of buprenorphine and its ≤1% w/w Sodium Azide. Liquid reagents contain ≤0.5% Bovine Serum, ≤0.2% Sodium Azide, metabolites in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to and ≤0.1% Drug-Specific Antibody (Mouse). be used in laboratories and provides a simple and rapid analytical screening procedure to detect buprenorphine and its metabolites in human urine. The assay is designed for use with a The reagents are harmful if swallowed. number of clinical chemistry analyzers. H317 - May cause allergic skin reaction. The semi-quantitative mode is for the purpose of enabling laboratories to determine an H334 - May cause allergy or asthma symptoms or breathing difficulties if inhaled. appropriate dilution of the specimen for confirmation by a confirmatory method such as EUH032 - Contact with acids liberates very toxic gas. Liquid chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures. Avoid breathing mist or vapor. Contaminated work clothing should not be allowed out of the workplace. Wear protective gloves/eye protection/face protection. In case of inadequate The assay provides only a preliminary analytical test result. A more specific alternative ventilation wear respiratory protection. If on skin: Wash with plenty of soap and water. IF chemical method must be used to obtain a confirmed analytical result. Gas chromatography/ INHALED: If breathing is difficult, remove victim to fresh air and keep at rest in a position mass spectrometry (GC/MS) or Liquid chromatography/tandem mass spectrometry comfortable for breathing. If skin irritation or rash occurs: Get medical advice/attention. (LC-MS/MS) is the preferred confirmatory method.1 If experiencing respiratory symptoms: Call a POISON CENTER or a doctor/physician. Wash contaminated clothing before reuse. Dispose of contents/container to location in accordance Clinical and professional judgment should be applied to any drugs of abuse test result, with local/regional/national/international regulations. particularly when preliminary results are used. For In Vitro Diagnostic Use Only. In the case of accidental spill, clean and dispose of material according to your laboratory’s Summary and Explanation of the Test Standard Operating Procedure (SOP), local, and state regulations. Buprenorphine is a semi-synthetic opioid analgesic derived from thebaine, a minor component of opium. Buprenorphine is structurally similar to morphine. It is a partial agonist receptor In the case of damaged packaging on arrival, contact your technical support representative modulator.2 Buprenorphine has a longer duration of action than morphine and can be (refer to last page of this Package Insert). administered sublingually as an analgesic. Subutex®, a higher dose buprenorphine formulation, is widely used in Europe and elsewhere as a substitution treatment for opiate addiction.3-5 Reagent Preparation and Storage The FDA has approved the use of Subutex and Suboxone®, containing buprenorphine as the For preparation of the solutions, refer to the section below. Remove the kit from refrigerated active drug, for the treatment of opiate dependence in the US. Subutex and Suboxone are storage (2-8°C) immediately prior to preparation of the solutions. the first narcotic drugs available under the US Drug Abuse Treatment Act (DATA) of 2003 for the treatment of opiate dependence that can be prescribed in the US in a physician’s work Prepare the solutions in the following order to minimize possible contamination. place.6 It has also been shown that buprenorphine has abuse potential and may cause dependency. In addition, a number of deaths have been recorded as a result of overdose R2 Enzyme Donor Solution with intravenously injected buprenorphine in conjunction with other psychotropic drugs Connect Bottle 2a (ED reagent) to Bottle 2 (ED Reconstitution Buffer) using one of the enclosed such as benzodiazepines.7 Buprenorphine is metabolized primarily by N-dealkylation to form adapters. Mix by gentle inversion, ensuring that all the lyophilized material from Bottle 2a norbuprenorphine and by conjugation to form buprenorphine-glucuronide and is transferred into Bottle 2. Avoid the formation of foam. Detach Bottle 2a and adapter from norbuprenorphine-glucuronide.8 Bottle 2 and discard. Cap Bottle 2 and let stand approximately 5 minutes at room temperature (21-25°C). Mix again. Record the reconstitution date on the bottle label. Place the bottle The CEDIA Buprenorphine II Assay uses recombinant DNA technology to produce a directly into the reagent compartment of the analyzer or into refrigerated storage (2-8°C) and 9 unique homogeneous enzyme immunoassay system. The assay is based on the bacterial let stand 30 minutes before use. β-galactosidase enzyme (Escherichia coli), which has been genetically engineered into two inactive fragments. These fragments spontaneously re-associate to form fully active enzymes R1 Enzyme Acceptor Solution that, in the assay format, cleave a substrate, generating a color change that can be measured Connect Bottle 1a (EA reagent) to Bottle 1 (EA Reconstitution Buffer) using one of the enclosed spectrophotometrically at 570 nm. adapters. Mix by gentle inversion, ensuring that all the lyophilized material from Bottle 1a is transferred into Bottle 1. Avoid the formation of foam. Detach Bottle 1a and adapter from In this assay, the analyte in the sample competes with the analyte conjugated to the inactive Bottle 1 and discard. Cap Bottle 1 and let stand approximately 5 minutes at room temperature fragment (enzyme donor) of the β-galactosidase enzyme for the antibody binding site. If the (21-25°C). Mix again. Record the reconstitution date on the bottle label. Place the bottle analyte is present in the sample, it binds to the antibody, leaving the inactive enzyme fragment directly into the reagent compartment of the analyzer or into refrigerated storage (2-8°C) and free to form active enzyme. If the analyte is not present in the sample, antibody binds to let stand 30 minutes before use. the analyte conjugated on the inactive fragment, inhibiting the re-association of inactive -galactosidase fragments, and no active enzyme is formed. The amount of active enzyme β NOTE 1: The components supplied in this kit are intended for use as an integral unit. Do formed and resultant absorbance change is directly proportional to the amount of analyte not mix components from different lots. present in the sample. Reagents NOTE 2: Avoid cross-contamination of reagents by matching reagent caps to the proper reagent bottles. The R2 solution (Enzyme Donor) should be yellow-orange in color. A red or red- 1 EA Reconstitution Buffer purple color indicates that the reagent has been contaminated and must be discarded. Discard Contains buffer salts, mouse monoclonal anti-buprenorphine derivative antibody Reagents 1 or 2 if turbidity or precipitates are observed. 0.8 - 1.0 mg/L, stabilizer, and preservative. 1a EA Reagent Contains 0.171 g/L Enzyme Acceptor, buffer salts and preservative. NOTE 3: The R1 and R2 solutions must be at the reagent compartment storage temperature 2 ED Reconstitution Buffer of the analyzer before performing the assay. Refer to the analyzer specific application sheet Contains buffer salts, stabilizers, and preservatives for additional information. 2a ED Reagent Store reagents at 2-8°C. DO NOT FREEZE. Contains 0.175 mg/L Enzyme Donor conjugated to buprenorphine derivative, 1.67 g/L chlorophenol red-β-D-galactopyranoside, stabilizers, detergent and preservative. For shelf life of the unopened components, refer to the box or bottle labels for the expiration date. Additional Materials Required (sold separately): R1 Solution: 60 days refrigerated on analyzer or at 2-8°C. Kit Description R2 Solution: 60 days refrigerated on analyzer or at 2-8°C. 10021390 CEDIA Negative Calibrator II (1 x 7.5 mL) 10020799 CEDIA Buprenorphine II Calibrator 10 ng/mL (1 x 5 mL) 10020800 CEDIA Buprenorphine II Calibrator 20 ng/mL (1 x 5 mL) 10020801 CEDIA Buprenorphine II Calibrator 50 ng/mL (1 x 5 mL) 10020802 CEDIA Buprenorphine II Calibrator 100 ng/mL (1 x 5 mL) 10020804 CEDIA Buprenorphine II Low (7.5 ng/mL) and High (12.5 ng/mL) Controls (2 x 5 mL each) Specimen Collection and Handling Qualitative Study Analysis Collect urine specimens in plastic or glass containers. Care should be taken to preserve the Total Precision (n=80) chemical integrity of the urine sample from the time it is collected until the time it is assayed. Buprenorphine Spike % of Cutoff LC-MS/MS Immunoassay Number of Specimens kept at room temperature that do not receive initial test within 8 days10 of arrival at Concentration (10 ng/mL) (ng/mL) Results Determinations the laboratory should be placed into a secure refrigeration unit at 2-8°C for up to 30 days.11,12 (ng/mL) (Negative/Positive) For longer storage prior to analysis or for sample retention after analysis, urine specimens 0 -100% 0.00 80 80/0 may be stored at -20°C.13 Studies have shown buprenorphine analytes in urine are stable at -20°C up to 85 days.13 2.5 -75% 2.99 80 80/0 Laboratories following the SAMHSA mandatory guidelines should refer to SAMHSA “Short- 5 -50% 5.31 80 80/0 Term Refrigerated Storage” and “Long-Term Storage” requirements.14 7.5 -25% 7.63 80 80/0 To protect the integrity of the sample, do not induce foaming and avoid repeated freezing 10 100% 10.99 80 27/53 and thawing. An effort should be made to keep pipetted samples free of gross debris.