Test for Lp-PLA 2 Abbott ARCHITECT c8000 and ci8200 Chemistry Analyzer Application Sheet This procedure is to be used in conjunction with the package insert for Test for Lp-PLA 2. Read and follow the package insert instructions carefully. Failure to follow the instructions may result in inaccurate results. Reagent barcode labels are not provided. For reagent ordering information, contact your local distributor. For further technical information, refer to the specific analyzer s operating instructions. PROCEDURE: 1. Program a user defined test with the parameters listed below. For more detailed instructions refer to the analyzer operating instructions. 2. Transfer the Test for Lp-PLA 2 reagents into appropriate Abbott reagent bottles, making sure bubbles are removed. 3. Load the reagent bottles as described in the analyzer operating instructions. 4. Calibrate with calibrator set included in the test kit. Calibrators and controls are lot specific; use the calibrators and controls provided in the test kit. A sample calibration curve specific to this analyzer is noted below for your reference. Your calibrator absorbances and resulting curve should appear similar. NOTE: The maximum reportable value is 400 nmol/min/ml. Do not dilute samples with results above 400 nmol/min/ml to obtain a value. Report result as >400 nmol/min/ml Hemoglobin present in test samples at a concentration > 1.0 mg/ml (> 0.1 g/dl or > 1g/L) may interfere with the test results. Bilirubin present in test samples at a concentration > 0.1 mg/ml (> 10 mg/dl or > 171 μmol/l) may interfere with the test results. See Test for Lp-PLA 2 package insert for further information. Refer to the sections: Warnings & Precautions, Procedural Notes and Limitations. USER DEFINED PARAMETERS: Number: XXX Type: Assay availability: Photometric Enabled Version: Date: Run controls for onboard reagents by: Lot Time: Operator: Reaction definition Reagent / Sample Validity checks Reaction mode: Rate up Primary Secondary Read times Wavelength: 412 / 524 Main: 19-21 Last required read: Absorbance range: Sample blank type: 21 0.1 None 3.0 Flex: Color correction: - - User Defined Page 1 of 6
Test for Lp-PLA 2 Number: XXX Type: Assay availability: Photometric Enabled Version: Date: Run controls for onboard reagents by: Lot Time: Operator: Reaction definition Reagent / Sample Validity checks R1 R2 Reagent: Reagent volume: 120 30 Diluent: <None> Water volume: Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0 Diluent name: STANDARD: Sample Diluted sample Diluent Water Dilution factor 1:1.00 Default dilution Number: XXX Type: Assay availability: Photometric Enabled Version: Date: Run controls for onboard reagents by: Lot Time: Operator: Reaction definition Reagent / Sample Validity checks Reaction check: None Rate linearity: % User Defined Page 2 of 6
Test for Lp-PLA 2 method: Calibrator set: Calibrator level: Concentration: Blank: 1 0.0 Replicates: 2 Cal 1: Cal 2: 2 3 50.0 100.0 Cal 3: 4 250.0 Cal 4: 5 400.0 method: Calibrator: Calibrator level Sample Diluted sample Diluent Water Blank: 1 Cal 1: 2 Cal 2: 3 Cal 3: Cal 4: 4 5 Page 3 of 6
Test for Lp-PLA 2 method: intervals: Full interval: 672 (hours) type: Adjust type: None method: Blank absorbance range: Span: Span absorbance range: Expected cal factor: -0.1000 Blank 0.0000 0.0000 -- 2.0000 -- Blank -- 2.0000 Expected cal factor tolerance %: 0 Page 4 of 6
Test for Lp-PLA 2 >> >> Calibrator Set >> Details Details for calibrator set... Calibrator set: Number of levels: 5 Lot number: Expiration date: ü Default ASSAY CAL 1 CAL 2 CAL 3 CAL 4 CAL 5 CAL 6 0.0 50.0 100.0 250.0 400.0 CALIBRATION CURVE Calibrators Absorbance (OD/min) 0 0.0075 50 0.0904 100 0.1788 250 0.3688 400 0.5362 Absorbance (OD/min) Ac5vity Calibra5on on the Abbo; Architect 0.600 0.500 0.400 0.300 0.200 0.100 0.000 0 50 100 150 200 250 300 350 400 450 Ac5vity Calibrators Page 5 of 6
TM Test Kit PERFORMANCE CHARACTERISTICS Performance characteristics were established using the Abbott Architect ci8200. On-Board Analyzer Reagent Storage Open bottles of reagents stored in the refrigerated compartment on the analyzer should be stable for up to 4 weeks. Laboratories should verify on-board reagent stability on their own analyzers, under typical laboratory conditions. Sensitivity The clinical sensitivity of the assay is 10 nmol/min/ml as determined by the limit of quantitation (the lowest concentration with acceptable precision). Assay Precision Intra-assay and inter-assay variability were determined by testing three human serum samples and two buffer controls with Lp-PLA 2 activity distributed throughout the calibration range of the assay. The data are summarized below: Mean Concentration Lp-PLA 2 Intra-assay % CV (n=20) Inter-assay % CV (n=20) 68-311 0.3-1.0% 0.4-0.8% Linearity From three pairs of serum samples with known high or low Lp-PLA 2 activity levels, a dilution series was prepared for each pair combination to assess linearity. The measured values were compared to the expected values and linear regression was determined. Sample Pair High Sample Low Sample Linear Regression Slope Intercept R 2 1 285 80 1.02-6.31 0.999 2 337 80 1.03 4.34 0.993 3 301 79 1.03-6.72 0.999 Method Comparison Correlation studies were performed comparing the Test for Lp-PLA 2 to the values established on the Beckman Coulter AU400 (formerly the Olympus AU400). Human serum samples with Lp-PLA 2 activities ranging from 6 to 382 nmol/min/ml were tested. from linear regression analysis are shown below. Linear Regression Slope = 1.06 Intercept = -3.42 R 2 =0.999 Number of Samples = 40 Page 6 of 6 and the logo are registered trademarks of diadexus. 2012 all rights reseved. All other trademarks and copyrights are property of their respective owners. FMD-10-016 rev 01 349 Oyster Point Blvd. South San Francisco, CA 94080 www.plactest.com 1.877.752.2837 1.650.246.6400