INSTRUCTIONS FOR USE

Transcription

1 VITROS Chemistry Products Reagent Rx ONLY Intended Use For in vitro diagnostic use only. VITROS Chemistry Products Reagent is used on the VITROS 5,1 FS Chemistry System, the VITROS 4600 Chemistry System and the VITROS 5600 Integrated System to quantitatively measure LDL Cholesterol (LDLC) concentration in serum and plasma. Low Density Lipoprotein (LDL) cholesterol is used to evaluate the risk of developing coronary heart disease (CHD). The risk of CHD increases with higher LDL cholesterol concentrations. Summary and Explanation of the Test Low-density lipoprotein (LDL) is one of six major classes of circulating lipoproteins. LDL transports cholesterol to peripheral tissues for cell membrane biosynthesis, and to the liver and other tissues for steroid hormone synthesis and to hepatocytes for bile acids synthesis. 1 LDL cholesterol levels are directly related to the risk of developing coronary heart disease, and lowering the LDL cholesterol level in the blood is a primary target of various cholesterol-lowering therapeutic agents. 2 Principles of the Procedure The quantitative measurement of LDLC is performed using the VITROS Chemistry Products Reagent in conjunction with the VITROS Chemistry Products Calibrator Kit 19 and VITROS Chemistry Products FS Calibrator 1 on the VITROS 5,1 FS/4600 Chemistry Systems and the VITROS 5600 Integrated System. The VITROS Reagent is a dual chambered package containing stable, ready-to-use liquid reagents that are used in a two-step reaction to quantitatively measure LDLC. In the first step, with the addition of R1, non-ldl cholesterol (such as HDL, VLDL and Chylomicrons) is selectively eliminated by reaction with cholesterol esterase and cholesterol oxidase to form cholestenone and hydrogen peroxide. The peroxide generated is immediately scavenged by catalase. Addition of reagent 2 initiates the second step, in which catalase is immediately inactivated with sodium azide. Surfactants then aid in dissociation of cholesterol and cholesterol esters from LDL particles and promote reaction with cholesterol esterase and cholesterol oxidase. The peroxide byproduct reacts with TOOS and 4-aminoantipyrine in the presence of peroxidase to form a colored quinone dye, which is measured spectrophotometrically at 600 nm. Once a calibration has been performed for each reagent lot, the LDLC concentration in each unknown sample can be determined using the stored calibration curve and the measured absorbance obtained in the assay of the sample. Test Type and Conditions Test Type Endpoint VITROS System 5600, 4600, 5,1 FS 5600, 4600, 5,1 FS 5600, 4600, 5,1 FS Approximate Incubation Time Temperature Wavelength R1 Pre-incubation: 4.4 minutes 37 C (98.6 F) Not Applicable Incubation 1: 4.7 minutes 37 C (98.6 F) 600 nm Incubation 2: 5.2 minutes 37 C (98.6 F) 600 nm Reaction Sample Volume 2.7 µl Not all products and systems are available in all countries. Version 9.0 Pub. No. J21171_EN 1 of 11

2 INSTRUCTIONS FOR USE Warnings and Precautions Reaction Scheme Step 1: R1 addition - Elimination of cholesterol from chylomicrons, VLDL and HDL Non-LDL cholesterol esters Cholesterol esterase cholesterol + fatty acids Cholesterol + O 2 Cholesterol oxidase cholestenone + H 2 O 2 2 H 2 O 2 Catalase * 2H 2 O + O 2 Step 2: R2 addition - Specific measurement of LDLC LDL particles Surfactants cholesterol + cholesterol esters + protein Cholesterol esters Cholesterol esterase cholesterol + fatty acids Cholesterol + O 2 Cholesterol oxidase cholestenone + H 2 O 2 2 H 2 O aminoantipyrine + TOOS Peroxidase quinone pigment + 4 H 2 O * Catalase is inhibited in the second step by sodium azide in Reagent 2 Warnings and Precautions For in vitro diagnostic use only. Reagents WARNING: WARNING: WARNING: This product contains sodium azide. Disposal of product into sinks with copper or lead plumbing should be followed with plenty of water to prevent formation of potentially explosive metallic azides. This reagent contains bovine blood components. This product should be handled using the same precautions as with any other blood or blood-derived product. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and CLSI Guideline M29 3 For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the Instructions for Use for the appropriate VITROS product, or to other manufacturer s product literature. Reactive Ingredients Reagent 1 (R1): Cholesterol esterase (Pseudomonas sp. E.C ) 600 U/L; cholesterol oxidase: (Microorganism, E.C ) 500 U/L; catalase: (Corynebacterium glutamicum, E.C ) 1,200,000 U/L; surfactant (polyoxyethylene compound) 0.3 % (w/v); dye precursor (N-Ethyl-N-[2-hydroxy-3-sulfopropyl]-3-methylaniline [TOOS]) 2.0 mm Reagent 2 (R2): Peroxidase (Horseradish, E.C ) 5,000 U/L; 4-Aminoantipyrine 4.0 mm; catalase Inhibitor (sodium azide) 0.05 % (w/v); polyoxyethylene alkylphenyl ether 1.0 % (w/v) Other Ingredients Reagent 1 (R1): Buffer, inorganic salt, scavenger, protein, preservative, processed water Reagent 2 (R2): Buffer, preservative, processed water Reagent Handling Caution: Do not use reagent packs with damaged or incompletely sealed packaging. Inspect the packaging for signs of damage. 2 of 11 Pub. No. J21171_EN Version 9.0

3 Reagent Storage and Stability Be careful when opening the outer packaging with a sharp instrument so as to avoid damage to the individual product packaging. The reagents are supplied ready for use. Avoid agitation, which may cause foaming or the formation of bubbles. Reagent Preparation 1. Remove from refrigerated storage 2. Immediately load into Supply 3. IMPORTANT: Do not loosen or remove caps prior to loading. Reagent Storage and Stability VITROS Chemistry Products Reagent is stable until the expiration date on the carton when it is stored and handled as specified. Do not use beyond the expiration date. Reagent Storage Condition Stability Unopened Refrigerated 2 8 C, protect from Until expiration date light Opened On-analyzer System turned on 4 weeks On-analyzer System turned off 30 minutes Verify performance with quality control materials: If the system is turned off for more than 30 minutes. After reloading reagents that have been removed from Supply 3 and stored for later use. Specimen Collection, Preparation and Storage Specimens Recommended Serum Plasma: Heparin IMPORTANT: Certain collection devices have been reported to affect other analytes and tests 4. Owing to the variety of specimen collection devices available, Ortho-Clinical Diagnostics is unable to provide a definitive statement on the performance of its products with these devices. Confirm that your collection devices are compatible with this test. Specimens Not Recommended Plasma: EDTA Serum Specimen Collection and Preparation Collect specimens using standard laboratory procedures. 5, 6 Note: For details on minimum fill volume requirements, refer to the operating instructions for your system. Patient Preparation No special patient preparation is necessary. Special Precautions Refer to Sample Dilution for dilution instructions. Centrifuge specimens and remove the serum from the cellular material within two hours of collection. 7 Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, C (64 82 F), prior to analysis. Version 9.0 Pub. No. J21171_EN 3 of 11

4 INSTRUCTIONS FOR USE Testing Procedure Specimen Storage and Stability Storage Temperature Stability Room temperature C (64 82 F) 1 day Refrigerated C (36 46 F) 3 days Frozen 8-20 C ( -4 F) 3 weeks IMPORTANT: Avoid repeated freeze-thaw cycles. Testing Procedure Materials Provided VITROS Chemistry Products Reagent Materials Required but Not Provided VITROS Chemistry Products Calibrator Kit 19 VITROS Chemistry Products FS Calibrator 1 Quality control materials, such as VITROS Chemistry Products Performance Verifiers I or II VITROS Chemistry Products FS Diluent Pack 2 (BSA/Saline) Operating Instructions Check reagent inventories at least daily to ensure that quantities are sufficient for the planned workload. For additional information, refer to the operating instructions for your system. IMPORTANT: Sample Dilution Bring all fluids and samples to room temperature, C (64 82 F), prior to analysis. On-Analyzer Sample Dilution If LDLC concentrations exceed the system s measuring (reportable) range, refer to the system operating instructions for more information on the On-Analyzer Dilution Procedure. Use VITROS Chemistry Products FS Diluent Pack 2 (BSA/Saline) for the dilution. Manual Dilution If LDL concentrations exceed the system s measuring (reportable) range: 1. Dilute sample with an equal volume of saline using VITROS Chemistry Products FS Diluent Pack 2 (BSA/Saline). 2. Reanalyze 3. Multiply the results by the dilution factor (2) to obtain an estimate of the original sample s LDL concentration. IMPORTANT: If using the system in On-Analyzer Dilution Mode, do not manually dilute samples for analysis. Refer to the operating instructions for your system for more information on the On-Analyzer Dilution Procedure. Calibration Required Calibrators VITROS Chemistry Products Calibrator Kit 19 VITROS Chemistry Products FS Calibrator 1 Calibrator Preparation, Handling, and Storage Refer to the Instructions for Use for VITROS Chemistry Products Calibrator Kit 19 and VITROS Chemistry Products FS Calibrator 1. Calibration Procedure Refer to the operating instructions for your system. When to Calibrate Calibrate: When the reagent lot number changes. When critical system parts are replaced due to service or maintenance. 4 of 11 Pub. No. J21171_EN Version 9.0

5 Quality Control When government regulations require. For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS assay may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operating instructions for your system. Calculations Absorbance is measured at 600 nm after a fixed incubation time. Once a calibration has been performed for each reagent lot, LDLC concentration in the unknown samples can be determined using the stored calibration curve and the measured absorbance obtained in the assay of each sample. Validity of a Calibration Calibration parameters are automatically assessed by the system against a set of quality parameters detailed in the Review Assay Data screen (found via Options Review/Edit Calibrations Review Assay Data). Failure to meet any of the predefined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration. Measuring (Reportable) Range Conventional Units SI Units Alternate Units (mg/dl) (mmol/l) (g/l) For out-of-range samples, refer to Sample Dilution. Traceability of Calibration Values assigned to the VITROS Chemistry Products Calibrator Kit 19 and VITROS Chemistry Products FS Calibrator Kit 1 for low-density lipoprotein cholesterol are traceable to the CDC LDL-C Reference Measurement Procedure. 8, 9 Quality Control Quality Control Material Selection IMPORTANT: VITROS Chemistry Products Performance Verifiers are recommended for use with the VITROS Chemistry and VITROS Integrated Systems. Evaluate the performance of other commercial control fluids for compatibility with this assay before using for quality control. Control materials other than VITROS Chemistry Products Performance Verifiers may show a difference when compared with other LDLC methods if they: Depart from a true human matrix. Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials containing sodium azide. Quality Control Procedure Recommendations Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: After calibration. According to local regulations or at least once each day that the assay is being performed. After specified service procedures are performed. Refer to the operating instructions for your system. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline Third Edition 10 or other published guidelines. For additional information, refer to the operating instructions for your system. Quality Control Material Preparation, Handling, and Storage Refer to the Instructions for Use for VITROS Chemistry Products Performance Verifier I and II or to other manufacturer's product literature. Version 9.0 Pub. No. J21171_EN 5 of 11

6 INSTRUCTIONS FOR USE Results Results Reporting Units and Unit Conversion The VITROS 5,1 FS/4600 Chemistry and VITROS Integrated Systems may be programmed to report results in conventional, SI, and alternate units. Conventional Units SI Units Alternate Units mg/dl mmol/l (mg/dl x ) g/l (mg/dl x 0.01) Limitations of the Procedure Known Interferences None identified. Other Limitations Certain drugs and clinical conditions are known to alter LDLC concentration in vivo. For additional information, refer to one of the published summaries. 12, 13, 14 For samples that are turbid and generate a Sample Integrity T-index flag with the VITROS Integrated and VITROS 5,1 FS/4600 Chemistry Systems, dilute the sample and reanalyze. Refer to Sample Dilution. For samples that contain triglyceride concentrations greater than 750 mg/dl, dilute the sample and reanalyze. Refer to Sample Dilution. Biased results may be observed with quality control and proficiency fluids containing sodium azide. Interpretation of Results and Expected Results Classification Low density lipoprotein cholesterol levels are categorized according to the classification scheme in the ATP III guidelines recommended by NCEP for samples collected from fasting patients. 11 Classification Conventional Units (mg/dl) SI Units (mmol/l) Alternate Units (g/l) Optimal < 100 < 2.59 < 1.00 Near to above Optimal Borderline High High Very High Performance Characteristics Method Comparison The plots and data below show the results of a comparison of serum samples analyzed on the VITROS 5,1 FS Chemistry System with those analyzed on a commercially available system. The VITROS assay has not been certified by the CDC Lipid Standardization Program for manufacturers. The table also shows the results of comparisons with serum and plasma samples on the VITROS 5600 Integrated System and the VITROS 5,1 FS Chemistry System. The testing followed NCCLS Protocol EP of 11 Pub. No. J21171_EN Version 9.0

7 Performance Characteristics Conventional Units SI Units VITROS 5,1 FS Chemistry System (mg/dl) VITROS 5,1 FS Chemistry System (mmol/l) Comparative Method (mg/dl) Comparative Method (mmol/l) n Slope Correlation Coefficient Conventional Units (mg/dl) Range of Sample Conc. Intercept Sy.x SI Units (mmol/l) Range of Sample Conc. Intercept Sy.x 5,1 FS vs. comparative method * vs. 5,1 FS * Homogeneous enzymatic assay Analytical processing hardware and software algorithms on the VITROS 4600 Chemistry System are designed to the same specifications as those applied to the VITROS 5,1 FS Chemistry System. Assay performance on the VITROS 4600 System has been demonstrated to be comparable to that on the VITROS 5,1 FS System. All performance characteristics for VITROS 5,1 FS System are therefore applicable to the VITROS 4600 System. Precision Precision was evaluated with quality control materials on the VITROS 5,1 FS System following NCCLS Protocol EP5. 16 Precision was also evaluated with quality control materials on the VITROS 5600 Integrated System following NCCLS Protocol EP5. 17 These results are guidelines. Variables such as instrument maintenance, environment, reagent storage/ handling, control material reconstitution, and sample handling can affect the reproducibility of test results. 5,1 FS 5600 Specificity Conventional Units (mg/dl) Mean Conc. Within Day SD * Within Lab SD ** Mean Conc. SI Units (mmol/l) Within Day SD * Within Lab SD ** Within Lab CV% ** No. Observ. No. Days * Within Day precision was determined using two-runs/ day with two replications per run. ** Within Lab precision was determined using a single lot of reagents and calibrating weekly (four calibration events). Analytical processing hardware and software algorithms on the VITROS 4600 Chemistry System are designed to the same specifications as those applied to the VITROS 5,1 FS Chemistry System. Assay performance on the VITROS 4600 System has been demonstrated to be comparable to that on the VITROS 5,1 FS System. All performance characteristics for VITROS 5,1 FS System are therefore applicable to the VITROS 4600 System. Substances that Do Not Interfere The substances listed in this table were tested on the VITROS Chemistry Products Reagent at LDL nominal concentrations of approximately 115 mg/dl (2.97 mmol/l) and found not to interfere, bias < 10 mg/dl (0.26 mmol/l), at the concentrations shown. Compound Concentration Acetaminophen 200 mg/l 1.3 mmol/l Acetylsalicylic acid 500 mg/l 2.8 mmol/l Allopurinol 10 mg/l 74 µmol/l Ampicillin 600 mg/l 1.7 mmol/l Version 9.0 Pub. No. J21171_EN 7 of 11

8 INSTRUCTIONS FOR USE References Compound Concentration Ascorbic acid 3 mg/dl 0.17 mmol/l Bilirubin 60 mg/dl 1.03 mmol/l Carbamazepine 20 mg/l 85 µmol/l Chloramphenicol 20 mg/l 62 µmol/l Chlordiazepoxide 20 mg/l 60 µmol/l Doxepine 10 mg/l 32 µmol/l D-Penicillamine 55 mg/l 370 µmol/l Erythromycin 400 mg/l 545 µmol/l Furosemide 30 mg/l 91 µmol/l Gentamicin 20 mg/l 42 µmol/l Hemoglobin 1000 mg/dl 10 g/l Methotrexate 0.45 mg/l 1 µmol/l Phenobarbital 150 mg/l 646 µmol/l Tetracycline 40 mg/l 90 µmol/l Tolbutamide 300 mg/l 1.1 mmol/l Valproic acid 300 mg/l 2.1 mmol/l References 1. Burtis C, Ashwood, ER (ed). Tietz Textbook of Clinical Chemistry, 3rd ed. Philadelphia, PA; WB Saunders Co; ; Executive Summary of the Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). JAMA 285: ; CLSI. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline Fourth Edition. CLSI document M29-A4. Wayne, PA: Clinical and Laboratory Standards Institute; Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86 90; CLSI. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture; Approved Standard-Sixth Edition. CLSI document H3-A6 (ISBN ). CLSI, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania USA; NCCLS. Procedures and Devices for the Collection of Diagnostic Capillary Blood Specimens; Approved Standard Fifth Edition. NCCLS document H4-A5 [ISBN ]. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; Bachorik PS, Ross JW. National Cholesterol Education Program Recommendations for Measurement of Low-Density Lipoprotein Cholesterol: Executive Summary. Clin. Chem. 41: ; National Cholesterol Education Program. Recommendations on lipoprotein measurement (NIH Publication No ). Bethesda, MD: National Heart, Lung, and Blood Institute CLSI. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline Third Edition. CLSI document C24-A3 (ISBN ). CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; NCEP. Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III), Final Report. NIH Publication No : II-7, III-6, National Institutes of Health. Bethesda. Maryland: September Young DS. Effects of Drugs on Clinical Laboratory Tests. Ed. 4 Washington, D.C.: AACC Press; Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.; AACC Press; Tryding N, Tufvesson C, Sonntag O (eds.) Drug Effects in Clinical Chemistry. Ed. 7. Stockholm: The National Corporation of Swedish Pharmacies, Pharmasoft AB, Swedish Society for Clinical Chemistry; NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline- Second Edition. NCCLS document EP9-A2 (ISBN ). CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; NCCLS. Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline. NCCLS document EP5-A [ISBN X]. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; NCCLS. Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline Second Edition. NCCLS document EP5-A2 [ISBN ]. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; of 11 Pub. No. J21171_EN Version 9.0

9 Glossary of Symbols Glossary of Symbols Revision History Date of Revision Version Description of Technical Changes* Updated EC Representative address. Added USA to legal manufacture address Prescription Use Statement added Warnings and Precautions: updated to align with the new Safety Data Sheets References: updated M29 Glossary of Symbols: added Globally Harmonized Symbols to comply with the Classification, Labelling and Packaging (CLP) Regulations Traceability of Calibration: Changed Reference Method to CDC LDL-C Reference Measurement Procedure Interpretation of Results and Expected Results: SI Units updated for Near to above Optimal, Borderline High and High Sign changed from > to for Very High References: Added reference Glossary of Symbols: Added Date of Manufacture Version 9.0 Pub. No. J21171_EN 9 of 11

10 INSTRUCTIONS FOR USE Revision History Date of Revision Version Description of Technical Changes* Reagent Handling Removed Store reagent packs at 2 8 C in a closed carton, protected from light. Interpretation of Results and Expected Results Updated References Updated Glossary of Symbols Updated Added information for the VITROS 4600 Chemistry System Added information for the VITROS 5600 Integrated System Test Type and Conditions Added statement Method Comparison Added information on sample types; updated NCEP statement References Updated Glossary of Symbols Updated Minor wording and formatting changes Reagent Handling: added new caution statement Reagent Storage and Stability: revised storage condition to add protected from light First release of document * The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate. Signature Obsolete Date 10 of 11 Pub. No. J21171_EN Version 9.0

11 Revision History Ortho-Clinical Diagnostics Felindre Meadows Pencoed Bridgend CF35 5PZ United Kingdom Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY USA VITROS is a registered trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., Version 9.0 Pub. No. J21171_EN 11 of 11