03032574122V2

APOBT
Tina-quant Apolipoprotein B ver.2
• Indicates cobas c systems on which reagents can be used
Order information Roche/Hitachi cobas c systems
Tina-quant Apolipoprotein B ver.2 cobas c 501
100 tests Cat. No. 03032574 122 System-ID 07 6569 4 •
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%

Shelf life at 2-8°C: See expiration date on
System information cobas c pack label.
APOBT: ACN 151 On-board in use and refrigerated on the analyzer: 12 weeks
Intended use Specimen collection and preparation
In vitro test for the quantitative determination of apolipoprotein B in human For specimen collection and preparation, only use suitable
serum and plasma on Roche/Hitachi cobas c systems. tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Summary1,2
Apolipoproteins are the protein constituents of the lipoproteins. The Serum.
lipoproteins are classified according to their ultracentrifugal flotation density. Plasma: Li-heparin, K2-EDTA plasma.
The liver synthesizes very low density lipoproteins (VLDL) which mainly The sample types listed were tested with a selection of sample collection
contain triglycerides and cholesterol. In the presence of lipoprotein lipase, tubes that were commercially available at the time of testing, i.e. not
the triglycerides are hydrolyzed and LDL particles with a high proportion of all available tubes of all manufacturers were tested. Sample collection
cholesterol are formed. Apolipoprotein B is the major protein constituent of systems from various manufacturers may contain differing materials
LDL. About one third of the LDL particles provide cholesterol to peripheral which could affect the test results in some cases.
cells. The other two thirds are metabolized by the liver. LDL-uptake in all of When processing samples in primary tubes (sample collection systems),
these tissues occurs via LDL receptors. Apolipoprotein B levels increase in follow the instructions of the tube manufacturer.
pregnancy, hypercholesterolemia, LDL receptor defects, bile obstruction, type
Centrifuge samples containing precipitates before performing the assay.
II hyperlipidemia and nephrotic syndrome. Apolipoprotein B levels decrease
during liver disease, α-β lipoproteinemia, sepsis and estrogen administration. Stability: 1 day at 15-25°C7
The combined determination of apolipoprotein A-I/apolipoprotein B and 8 days at 2-8°C7
the calculation of the apolipoprotein B : apolipoprotein A-I ratio can reflect 2 months at (-15)-(-25)°C8 (only freeze once)
a lipid metabolism disorder and the risk of developing atherosclerosis
or coronary heart disease particularly well, thus providing an excellent Materials provided
addition to the classical HDL/LDL-cholesterol determination. A high level See “Reagents - working solutions” section for reagents.
of apolipoprotein A-I (HDL) and a low level of apolipoprotein B (LDL)
correlate best with a low risk for these diseases. Materials required (but not provided)
See “Order information” section.
Test principle3,4,5,6 Distilled water
Immunoturbidimetric assay. General laboratory equipment
Anti-apolipoprotein B antibodies react with the antigen in the sample
to form antigen/antibody complexes which, following agglutination, Assay
can be measured turbidimetrically. For optimal performance of the assay, follow the directions given in this
document for the analyzer concerned. Refer to the appropriate operator
Reagents - working solutions manual for analyzer-specific assay instructions.
The performance of applications not validated by Roche is not
R1 TRIS buffer: 50 mmol/L, pH 8.0; PEG: 4.2%; detergent; preservative
warranted and must be defined by the user.
R2 Anti-human apolipoprotein B antibodies (sheep): dependent on titer;
TRIS buffer: 100 mmol/L, pH 8.0; preservative Application for serum and plasma
Precautions and warnings cobas c 501 test definition
For in vitro diagnostic use. Assay type 2 Point End
Exercise the normal precautions required for handling all laboratory reagents. Reaction time / Assay points 10 / 10-46
Safety data sheet available for professional user on request. Wavelength (sub/main) 700/340 nm
Disposal of all waste material should be in accordance with local guidelines.
Reaction direction Increase
Reagent handling Units g/L (µmol/L, mg/dL)
Ready for use. Reagent pipetting Diluent (H2O)
R1 100 µL –
Storage and stability R2 25 µL 30 µL
APOBT Sample volumes Sample Sample dilution
Shelf life at 2-8°C: See expiration date on Sample Diluent (NaCl)
cobas c pack label. Normal 6 µL 9 µL 180 µL
On-board in use and refrigerated on the analyzer: 12 weeks Decreased 6 µL 9 µL 180 µL
Increased 12 µL 9 µL 180 µL

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APOBT
Tina-quant Apolipoprotein B 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 7-18 years 0.28-1.14 g/L (1.06-4.32 µmol/L, 28-114 mg/dL)
Multiply the lot-specific C.f.a.s. Lipids calibrator Men 0.66-1.33 g/L (2.50-5.04 µmol/L, 66-133 mg/dL)
value by the factors below to determine the standard Women 0.60-1.17 g/L (2.27-4.43 µmol/L, 60-117 mg/dL)
concentrations for the six-point calibration curve: IFCC, risk related <1.00 g/L (<3.79 µmol/L, <100 mg/dL)
S2: 0.323 S5: 2.100 target value
S3: 0.600 S6: 3.500
Each laboratory should investigate the transferability of the expected values to
S4: 1.617 its own patient population and if necessary determine its own reference ranges.
Calibration mode RCM
Calibration frequency Full calibration Specific performance data
- after reagent lot change Representative performance data on the analyzers are given below.
Results obtained in individual laboratories may differ.
- and as required following quality control
procedures Precision
Traceability: This method has been standardized against the IFCC SP3-07 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).
The following results were obtained:
Quality control
Within-run Mean SD CV
For quality control, use control materials as listed in the
g/L g/L %
“Order information” section.
(µmol/L, mg/dL) (µmol/L, mg/dL)
Other suitable control material can be used in addition.
The control intervals and limits should be adapted to each laboratory’s Precinorm L 0.82 (3.11, 82) 0.01 (0.04, 1) 1.8
individual requirements. Values obtained should fall within the defined Precipath L 1.75 (6.63, 175) 0.02 (0.08, 2) 1.4
limits. Each laboratory should establish corrective measures to be Human serum 1 0.83 (3.15, 83) 0.01 (0.04, 1) 1.1
taken if values fall outside the limits. Human serum 2 1.23 (4.66, 123) 0.01 (0.04, 1) 1.1
Calculation Total Mean SD CV
Roche/Hitachi cobas c systems automatically calculate the analyte g/L g/L %
concentration of each sample. (µmol/L, mg/dL) (µmol/L, mg/dL)
Conversion factors: g/L x 3.79a = µmol/L13
Precinorm L 0.83 (3.15, 83) 0.03 (0.11, 3) 3.1
g/L x 100 = mg/dL
Precipath L 1.86 (7.05, 186) 0.04 (0.15, 4) 2.3
a) measured as B48
Human serum 3 0.87 (3.30, 87) 0.03 (0.11, 3) 2.9
Limitations - interference14
Human serum 4 2.34 (8.87, 234) 0.06 (0.23, 6) 2.7
Criterion: Recovery within ±10% of initial values at apolipoprotein B
levels of 1.00 g/L (3.79 µmol/L, 100 mg/dL). Method comparison
Icterus: No significant interference up to an I index of 60 (approximate Apolipoprotein B values for human serum and plasma samples obtained
conjugated and unconjugated bilirubin concentration: 1026 µmol/L (60 mg/dL)). on a Roche/Hitachi cobas c 501 analyzer (y) were compared with those
Hemolysis: No significant interference up to an H index of 1000 (approximate determined using the same reagent on a Roche/Hitachi 917 analyzer (x).
hemoglobin concentration: 621 µmol/L (1000 mg/dL)). Sample size (n) = 123
Lipemia (Intralipid): No significant interference up to an L index of Passing/Bablok17 Linear regression
1000. There is poor correlation between the L index (corresponds y = 1.000x + 0.05 g/L y = 0.995x + 0.05 g/L
to turbidity) and triglycerides concentration. τ = 0.960 r = 0.997
Rheumatoid factors ≤ 1200 IU/mL do not interfere. The sample concentrations were between 0.22 and 3.78 g/L
No high-dose hook effect was observed up to an apolipoprotein B (0.83 and 14.3 µmol/L, 22 and 378 mg/dL).
concentration of 9.0 g/L (34.1 µmol/L, 900 mg/dL).
References
Drugs: No interference was found using common drug panels.15
1. Riesen WF. Apolipoproteine. In: Thomas L, ed. Labor und
In very rare cases gammopathy, in particular type IgM (Waldenström’s Diagnose, 5th ed. Frankfurt 1998:171-190.
macroglobulinemia), may cause unreliable results. 2. Brewer HB, Gregg RE, Hoeg JM, Fojo SS. Apolipoproteins and
For diagnostic purposes, the results should always be assessed in conjunction Lipoproteins in Human Plasma: an overview. Clin Chem 1988;34:B4-B8.
with the patient’s medical history, clinical examination and other findings. 3. Becker W, Rapp W, Schwick HG, Störiko K. Methoden zur quantitativen
Special wash requirements Bestimmung von Plasmaproteinen durch Immunpräzipitation.
The determination of certain analytes interferes with this assay requiring Z Klin Chem Klin Biochem 1968;6:113-122.
a special wash step. Refer to the NaOHD/SMS/Multiclean method sheet 4. Siedel J et al. Immunoturbidimetric Method for Routine Determinations of
and the operator manual for further instructions. Apolipoproteins B, BI and B in Normo- and Hyperlipemic Sera compared
Measuring range with Immunonephelometry. Clin Chem 1988;34:1821-1825.
0.2-4.0 g/L (0.76-15.2 µmol/L, 20-400 mg/dL) 5. Rifai N, King ME. Immunoturbidimetric Assays of Apolipoproteins A,
AI, AII and B in serum. Clin Chem 1986;32:957-961.
Extended measuring range (calculated)
6. Naito HK. Reliability of Lipid, Lipoprotein and Apolipoprotein
0.1-4.0 g/L (0.38-15.2 µmol/L, 10-400 mg/dL)
Measurements. Clin Chem 1988;34 B84-B94.
Lower detection limit
7. Use of Anticoagulants in Diagnostic Laboratory Investigations. WHO
0.03 g/L (0.11 µmol/L, 3 mg/dL).
Publication WHO/DIL/LAB/99.1 Rev.2. 2002.
The lower detection limit represents the lowest measurable analyte
8. Evans K, Mitcheson J, Laker M. Effect of Storage at 4°C and
level that can be distinguished from zero. It is calculated as the value
-20°C on Lipid, Lipoprotein, and Apolipoprotein Concentrations.
lying three standard deviations above that of the lowest standard
Clin Chem 1995;41:392-396.
(standard 1 + 3 SD, within-run , n = 21).
9. Marcovina SM, Albers JJ, Dati F, Ledue TB, Ritchie RF. International
Federation of Clinical Chemistry Standardization Project for Measurements
of Apolipoproteins A-I and B. Clin Chem 1991;37:1676-1682.

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03032574122V2

APOBT
Tina-quant Apolipoprotein B ver.2
10. Albers JJ, Marcovina SM, Kennedy H. International Federation of
Clinical Chemistry Standardization Project for Measurements of
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 B 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.
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will be free from defects in material and workmanship until the expiration
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2006-08, V 2 English 3/3 cobas c systems