APOAT Tina-quant Apolipoprotein A-1 ver.2 • Indicates cobas c systems on which reagents can be used Order information Roche/Hitachi cobas c systems Tina-quant Apolipoprotein A-1 ver.2 cobas c 501 100 tests Cat. No. 03032566 122 System-ID 07 6568 6 • Calibrator f.a.s. Lipids (3 x 1 mL) Cat. No. 12172623 122 Code 424 Calibrator f.a.s. Lipids (3 x 1 mL, for USA) Cat. No. 12172623 160 Code 424 Precinorm L (4 x 3 mL) Cat. No. 10781827 122 Code 304 Precipath L (4 x 3 mL) Cat. No. 11285874 122 Code 305 NaCl Diluent 9% (50 mL) Cat. No. 04489357 190 System-ID 07 6869 3
English NaCl Diluent 9%
System information Shelf life at 2-8°C: See expiration date on APOAT: ACN 168 cobas c pack label. On-board in use and refrigerated on the analyzer: 12 weeks Intended use In vitro test for the quantitative determination of Apolipoprotein A-1 in human Specimen collection and preparation serum and plasma on Roche/Hitachi cobas c systems. For specimen collection and preparation, only use suitable tubes or collection containers. Summary1,2 Only the specimens listed below were tested and found acceptable. Apolipoproteins are the protein constituents of the lipoproteins. The Serum. lipoproteins are classified according to their ultracentrifugal flotation Plasma: Li-heparin, K2-EDTA plasma. density. Apolipoprotein A-1 is the major protein constituent of high-density lipoproteins (HDL). HDL are synthesized by the intestines and the liver. The sample types listed were tested with a selection of sample collection They transport excess cellular cholesterol from extrahepatic tissue and tubes that were commercially available at the time of testing, i.e. not peripheral cells to the liver. Additionally, apolipoprotein A-1 activates the all available tubes of all manufacturers were tested. Sample collection enzyme lecithin-cholesterol-acyltransferase (LCAT), which catalyzes the systems from various manufacturers may contain differing materials esterification of cholesterol, thereby enhancing the lipid-carrying capacity which could affect the test results in some cases. of the lipoproteins. Apolipoprotein A-1 levels increase in liver disease, When processing samples in primary tubes (sample collection systems), pregnancy and as a result of estrogen administration (e.g. oral contraceptives). follow the instructions of the tube manufacturer. Apolipoprotein A-1 levels decrease in inherited hypo-α-lipoproteinemia Centrifuge samples containing precipitates before performing the assay. (e.g. Tangier disease), cholestasis, sepsis and atherosclerosis. The liver also synthesizes very low density lipoproteins (VLDL) which mainly contain Stability: 1 day at 15-25°C7 triglycerides and cholesterol. In the presence of lipoprotein lipase the 3 days at 2-8°C7 triglycerides are hydrolyzed and LDL-particles with a high proportion of 2 months at (-15)-(-25)°C8 (only freeze once) cholesterol are formed. Apolipoprotein B is the main constituent of LDL. The combined determination of apolipoprotein A-1/apolipoprotein B and Materials provided the calculation of the apolipoprotein B : apolipoprotein A-1 ratio can reflect See “Reagents - working solutions” section for reagents. a lipid metabolism disorder and the risk of developing atherosclerosis or coronary heart disease particularly well, thus providing an excellent Materials required (but not provided) addition to the classical HDL/LDL-cholesterol determination. A high level See “Order information” section. of apolipoprotein A-1 (HDL) and a low level of apolipoprotein B (LDL) Distilled water correlate best with a low risk for these diseases. General laboratory equipment
Test principle3,4,5,6 Assay
Immunoturbidimetric assay. For optimal performance of the assay, follow the directions given in this Anti-apolipoprotein A-1 antibodies react with the antigen in the sample document for the analyzer concerned. Refer to the appropriate operator to form antigen/antibody complexes which, following agglutination, manual for analyzer-specific assay instructions. can be measured turbidimetrically. The performance of applications not validated by Roche is not warranted and must be defined by the user. Reagents - working solutions R1 TRIS buffer: 50 mmol/L, pH 8.0; PEG: 3.8%; detergent; preservative Application for serum and plasma R2 Anti-human apolipoprotein A-1 antibodies (sheep): dependent on titer; cobas c 501 test definition TRIS buffer: 100 mmol/L, pH 8.0; preservative Assay type 2 Point End Reaction time / Assay points 10 / 10-28 Precautions and warnings Wavelength (sub/main) 700/340 nm For in vitro diagnostic use. Reaction direction Increase Exercise the normal precautions required for handling all laboratory reagents. Safety data sheet available for professional user on request. Units g/L (µmol/L, mg/dL) Disposal of all waste material should be in accordance with local guidelines. Reagent pipetting Diluent (H2O) R1 100 µL – Reagent handling R2 25 µL 30 µL Ready for use Sample volumes Sample Sample dilution Storage and stability Sample Diluent (NaCl) APOAT Normal 3 µL 9 µL 180 µL Shelf life at 2-8°C: See expiration date on Decreased 3 µL 9 µL 180 µL cobas c pack label. On-board in use and refrigerated on the analyzer: 12 weeks Increased 6 µL 9 µL 180 µL
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APOAT Tina-quant Apolipoprotein A-1 ver.2 Calibration Expected values16 Calibrators S1: H2O The following values were obtained using serum from healthy subjects: S2-S6: C.f.a.s. Lipids 17-18 years 0.32-2.27 g/L (11.4-81.0 µmol/L, 32-227 mg/dL) Multiply the lot-specific C.f.a.s. Lipids calibrator values by Men 1.04-2.02 g/L (37.1-72.1 µmol/L, 104-202 mg/dL) the factors below to determine the standard concentrations Women 1.08-2.25 g/L (38.6-80.3 µmol/L, 108-225 mg/dL) for the six-point calibration curve: IFCC, risk related >1.15 g/L (>41.1 µmol/L, >115 mg/dL) S2: 0.208 S5: 1.313 target value S3: 0.412 S6: 2.006 Each laboratory should investigate the transferability of the expected values to S4: 1.000 its own patient population and if necessary determine its own reference ranges. Calibration mode RCM Specific performance data Calibration Full calibration Representative performance data on the analyzers are given below. frequency - after reagent lot change Results obtained in individual laboratories may differ. - and as required following quality control procedures Precision Traceability: This method has been standardized against the IFCC SP1-01 Reproducibility was determined using human samples and controls in reference standard (WHO-IRP October 1992).9,10,11,12 an internal protocol (within-run n = 21, total n = 63). Quality control The following results were obtained: For quality control, use control materials as listed in the Within-run Mean SD CV “Order information” section. g/L (µmol/L) g/L (µmol/L) % Other suitable control material can be used in addition. Precinorm L 1.60 (57.1) 0.02 (0.7) 1.1 The control intervals and limits should be adapted to each laboratory’s Precipath L 1.00 (35.7) 0.01 (0.4) 1.5 individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be Human serum 1 0.99 (35.3) 0.02 (0.7) 1.5 taken if values fall outside the limits. Human serum 2 2.59 (92.5) 0.02 (0.7) 1.0
Calculation Total Mean SD CV
Roche/Hitachi cobas c systems automatically calculate the analyte g/L (µmol/L) g/L (µmol/L) % concentration of each sample. Precinorm L 1.74 (62.1) 0.08 (2.9) 4.7 Conversion factors: g/L x 35.7 = µmol/L Precipath L 1.06 (37.8) 0.04 (1.4) 3.9 g/L x 100 = mg/dL Human serum 3 1.17 (41.8) 0.04 (1.4) 3.6 mg/dL x 0.01 = g/L Human serum 4 2.40 (85.7) 0.03 (1.1) 1.4 Limitations - interference14 Method comparison Criterion: Recovery within ±10% of initial values at apolipoprotein A-1 Apolipoprotein A-1 values for human serum and plasma samples obtained levels of 1.00 g/L (35.7 µmol/L). on a Roche/Hitachi cobas c 501 analyzer (y) were compared with those Icterus: No significant interference up to an I index of 60 (approximate determined using the same reagent on a Roche/Hitachi 917 analyzer (x). conjugated and unconjugated bilirubin concentration: 1026 µmol/L (60 mg/dL)). Sample size (n) = 126 Hemolysis: No significant interference up to an H index of 1000 (approximate Passing/Bablok17 Linear regression hemoglobin concentration: 621 µmol/L (1000 mg/dL)). y = 1.014x + 0.07 g/L y = 1.006x + 0.09 g/L Lipemia (Intralipid): No significant interference up to an L index of τ = 0.949 r = 0.996 1000. There is poor correlation between the L index (corresponds The sample concentrations were between 0.38 and 3.34 g/L (13.6 and to turbidity) and triglycerides concentration. 119 µmol/L). Rheumatoid factors ≤ 1200 IU/mL do not interfere. References No high-dose hook effect was observed up to an apolipoprotein A-1 1. Riesen WF. Apolipoproteine. In: Thomas L, ed. Labor und concentration of 11 g/L (392 µmol/L). Diagnose, 5th ed. Frankfurt 1998:171-190. Drugs: No interference was found using common drug panels.15 2. Brewer HB, Gregg RE, Hoeg JM, Fojo SS. Apolipoproteins and In very rare cases gammopathy, in particular type IgM (Waldenström’s Lipoproteins in Human Plasma: an overview. Clin Chem 1988;34:B4-B8. macroglobulinemia), may cause unreliable results. 3. Becker W, Rapp W, Schwick HG, Störiko K. Methoden zur quantitativen For diagnostic purposes, the results should always be assessed in conjunction Bestimmung von Plasmaproteinen durch Immunpräzipitation. with the patient’s medical history, clinical examination and other findings. Z Klin Chem Klin Biochem 1968;6:113-122. 4. Siedel J et al. Immunoturbidimetric Method for Routine Determinations Special wash requirements of Apolipoproteins A-I, A-II and B in Normo- and Hyperlipemic Sera The determination of certain analytes interferes with this assay requiring compared with Immunonephelometry. Clin Chem 1988;34:1821-1825. a special wash step. Refer to the NaOHD/SMS/Multiclean method sheet 5. Rifai N, King ME. Immunoturbidimetric Assays of Apolipoproteins A, and the operator manual for further instructions. AI, AII and B in serum. Clin Chem 1986;32:957-961. Measuring range 6. Naito HK. Reliability of Lipid, Lipoprotein and Apolipoprotein 0.2-4.0 g/L (7.14-143 µmol/L) Measurements. Clin Chem 1988;34:B84-B94. Extended measuring range (calculated) 7. Use of Anticoagulants in Diagnostic Laboratory Investigations. WHO 0.1-4.0 g/L (3.57-143 µmol/L) Publication WHO/DIL/LAB/99.1 Rev.2. 2002. 8. Evans K, Mitcheson J, Laker M. Effect of Storage at 4°C and Lower detection limit -20°C on Lipid, Lipoprotein, and Apolipoprotein Concentrations. 0.03 g/L (1.07 µmol/L) Clin Chem 1995;41:392-396. The lower detection limit represents the lowest measurable analyte 9. Marcovina SM, Albers JJ, Dati F, Ledue TB, Ritchie RF. International level that can be distinguished from zero. It is calculated as the value Federation of Clinical Chemistry Standardization Project for Measurements lying three standard deviations above that of the lowest standard of Apolipoproteins A-I and B. Clin Chem 1991;37:1676-1682. (standard 1 + 3 SD, within-run precision, n = 21). 10. Albers JJ, Marcovina SM, Kennedy H. International Federation of Clinical Chemistry Standardization Project for Measurements of
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APOAT Tina-quant Apolipoprotein A-1 ver.2 Apolipoproteins A-I and B. II. Evaluation and Selection of Candidate Reference Materials. Clin Chem 1992;38:658-662. 11. Marcovina SM, Albers JJ, Henderson LO, Hannon WH. International Federation of Clinical Chemistry Standardization Project for Measurements of Apolipoproteins A-I and B. III. Comparability of Apolipoprotein A-I Values by Use of International Reference Material. Clin Chem 1993;39:773-781. 12. Marcovina SM et al. International Federation of Clinical Chemistry Standardization Project for Measurements of Apolipoproteins A-I and B. IV. Comparability of apolipoprotein B Values by Use of International Reference Material. Clin Chem 1994;40:586-592. 13. Young DS, Huth EJ. SI Units For Clinical Measurement. American College of Physicians, 1998. 14. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-474. 15. Report on the Symposium “Drug effects in clinical chemistry methods”, Breuer J, Eur J Clin Chem Clin Biochem 1996;34:385-386. 16. Heil W, Koberstein R, Zawta B. Reference Ranges for Adults and Children; Pre-analytical Considerations. Published by Roche Diagnostics 2004. 17. Bablok W et al. A General Regression Procedure for Method Transformation. J Clin Chem Clin Biochem 1988;26:783-790. FOR US CUSTOMERS ONLY: LIMITED WARRANTY Roche Diagnostics warrants that this product will meet the specifications stated in the labeling when used in accordance with such labeling and will be free from defects in material and workmanship until the expiration date printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE FOR INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL DAMAGES.
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