Assay Summary: Sample Type Serum Sample Volume 50 L Calibrator D Sensitivity and Assay Range 0.5-100 NG/ML ( G/L)
Assay Summary: Sample Type Serum Sample Volume 50 L Calibrator D Sensitivity and Assay Range 0.5-100 NG/ML ( G/L)
CEA
Assay Summary
Sample Type Serum
Sample Volume 50 µL
Calibrator D
Sensitivity and Assay Range 0.5–100 ng/mL (µg/L)
Contents
REF Contents Number of Tests
09788458 5 ReadyPack® primary reagent packs containing ADVIA Centaur® 500
(110762) CEA Lite Reagent and Solid Phase
ADVIA Centaur CEA Master Curve card
or
00937450 1 ReadyPack primary reagent pack containing ADVIA Centaur 100
(110761) CEA Lite Reagent and Solid Phase
ADVIA Centaur CEA Master Curve card
For a definition of symbols used in product labeling, please refer to Understanding the Symbols
in Appendix D.
Intended Use
For in vitro diagnostic use in the quantitative measurement of carcinoembryonic antigen (CEA)
in serum to aid in the management of cancer patients in whom changing concentrations of CEA
are observed using the ADVIA Centaur and ADVIA Centaur XP systems.
WARNING: The concentration of CEA in a given specimen, as determined by assays from
different manufacturers, can vary due to differences in assay methods and reagent specificity.
The results reported by the laboratory to the physician must include the identity of the CEA
assay used. Values obtained with different assay methods cannot be used interchangeably. If, in
the course of monitoring a patient, the assay method used for determining serial CEA levels is
changed, the laboratory must perform additional serial testing to confirm baseline values.
United States federal law restricts this device to sale and distribution by or on the order of a
physician, or to a clinical laboratory; and use is restricted to, by, or on the order of a physician.
Optional Reagents
REF Description Contents
07584944 ADVIA Centaur CEA Diluent 2 ReadyPack ancillary reagent packs
(110322) containing 5 mL/pack
01989969 CEA Diluent 10 mL/vial
(672255)
672427 CEA Master Curve Material 6 x 1 mL
Assay Principle
The ADVIA Centaur CEA assay is a two-site sandwich immunoassay using direct
chemiluminometric technology, which uses constant amounts of two antibodies. The first
antibody, in the Lite Reagent, is a purified polyclonal rabbit anti-CEA antibody labeled with
acridinium ester. The second antibody, in the Solid Phase, is a monoclonal mouse anti-CEA
antibody covalently coupled to paramagnetic particles.
The system automatically performs the following steps:
• dispenses 50 μL of sample into a cuvette
• dispenses 50 μL of Lite Reagent and 250 μL of Solid Phase and incubates for 7.5 minutes
at 37°C
• separates, aspirates, and washes the cuvettes with reagent water8
• dispenses 300 μL each of Acid Reagent and Base Reagent to initiate the chemiluminescent
reaction
• reports results according to the selected option, as described in the system operating
instructions or in the online help system
• Tightly cap and refrigerate specimens at 2° to 8°C if the assay is not completed within
8 hours.
• Freeze samples at or below -20°C if the sample is not assayed within 48 hours.
• Freeze samples only once and mix thoroughly after thawing.
Before placing samples on the system ensure that:
• Samples are free of fibrin or other particulate matter. Remove particulates by centrifugation
at 1000 x g for 15 to 20 minutes.
• Samples are free of bubbles.
Reagents
Store the reagents upright at 2–8°C.
Mix all primary reagent packs by hand before loading them onto the system. Visually inspect the bottom of the
reagent pack to ensure that all particles are dispersed and resuspended. For detailed information about
preparing the reagents for use, refer to Appendix C, Handling Reagents.
Reagent Pack Reagent Volume Ingredients Storage Stability
ADVIA Centaur Lite Reagent 5.0 mL/ polyclonal rabbit anti-CEA 2–8°C until the expiration date on
CEA ReadyPack reagent antibody (~400 ng/mL) the pack label.
primary reagent pack labeled with acridinium ester For onboard stability, refer to
pack in phosphate buffered saline Onboard Stability and
with protein stabilizers, Calibration Interval.
sodium azide (0.12%), and
preservatives
Solid Phase 25.0 mL/ monoclonal mouse 2–8°C until the expiration date on
reagent anti-CEA antibody the pack label.
pack (~120 μg/mL) covalently For onboard stability, refer to
coupled to paramagnetic Onboard Stability and
particles in phosphate Calibration Interval.
buffered saline with protein
stabilizers, sodium azide
(0.11%), and preservatives
ADVIA Centaur CEA Diluent 5.0 mL/ bicine buffer, gelatin, and 2–8°C until the expiration date on
CEA reagent BSA with preservatives the pack label
ReadyPack pack and sodium azide (0.1%) or
ancillary reagent 28 consecutive days after
pack accessing the ancillary
reagent pack
R22 Harmful! Harmful if swallowed. After contact with skin, wash immediately with plenty of soap
S28 and water. Contains: sodium azide; Lite Reagent, Solid Phase
CAUTION: This device contains material of animal origin and should be handled as a potential
carrier and transmitter of disease.
NOTE: Sodium azide can react with copper and lead plumbing to form explosive metal azides.
On disposal, flush reagents with a large volume of water to prevent the buildup of azides, if
disposal into a drain is in compliance with federal, state, and local requirements.
For in vitro diagnostic use.
Loading Reagents
Ensure that the system has sufficient primary and ancillary reagent packs. For detailed
information about preparing the system, refer to the system operating instructions or to the
online help system.
Use this procedure to mix ADVIA Centaur CEA primary reagent packs that are unpierced.
CAUTION: Do not use this procedure for pierced ADVIA Centaur CEA reagent packs. Discard
pierced ADVIA Centaur CEA reagent packs that have been removed from the system.
1. Hold the reagent pack firmly with thumb on one side and fingers on the other side. Shake
vigorously for 15 seconds, using a back and forth motion.
15 sec.
2. Hold the reagent pack firmly at one end, with film side up, and tap sharply on a bench top
five times to reduce foaming caused by shaking.
x5
Visually inspect the bottom of the reagent pack to ensure that all particles are dispersed and
resuspended.
Load the unpierced ReadyPack reagent packs in the primary reagent area using the arrows as a
placement guide. The system automatically mixes the primary reagent packs to maintain
homogeneous suspension of the reagents. For detailed information about loading reagents, refer
to the system operating instructions or to the online help system.
If automatic dilution of a sample is required, load ADVIA Centaur CEA Diluent in the ancillary
reagent entry.
• Discard the primary reagent packs at the end of the onboard stability interval.
• Do not use reagents beyond the expiration date.
Quality Control
For detailed information about entering quality control values, refer to the system operating
instructions or to the online help system.
To monitor system performance and chart trends, as a minimum requirement, two levels of
quality control material should be assayed on each day that samples are analyzed. Quality
control samples should also be assayed when performing a two-point calibration. Treat all
quality control samples the same as patient samples.
Siemens Healthcare Diagnostics recommends the use of commercially available quality control
materials with at least 2 levels (low and high). A satisfactory level of performance is achieved
when the analyte values obtained are within the Acceptable Control Range for the system or
within your range, as determined by an appropriate internal laboratory quality control scheme.
If the quality control results do not fall within the Expected Values or within the laboratory’s
established values, do not report results. Take the following actions:
• Verify that the materials are not expired.
• Verify that required maintenance was performed.
• Verify that the assay was performed according to the instructions for use.
• Rerun the assay with fresh quality control samples.
• If necessary, contact your local technical support provider or distributor for assistance.
Sample Volume
This assay requires 50 μL of sample for a single determination. This volume does not include
the unusable volume in the sample container or the additional volume required when
performing duplicates or other tests on the same sample. For detailed information about
determining the minimum required volume, refer to Sample Volume Requirements in the
ADVIA Centaur Reference Manual.
NOTE: The sample volume required to perform onboard dilution differs from the sample volume
required to perform a single determination. Refer to the following information for the sample
volume required to perform onboard dilutions:
Dilution Sample Volume (µL)
1:5 40
1:10 40
1:50 40
1:100 40
Assay Procedure
For detailed procedural information, refer to the system operating instructions or to the online
help system.
Procedural Notes
Calculations
For detailed information about how the system calculates results, refer to the system operating
instructions or to the online help system.
The system reports serum CEA results in ng/mL (mass units) or μg/L (SI units), depending on
the units defined when setting up the assay. The conversion formula is 1 ng/mL = 1 μg/L.
Dilutions
• Serum samples with CEA levels greater than 100 ng/mL (μg/L) must be diluted and
retested to obtain accurate results.
• Patient samples can be automatically diluted by the system or prepared manually.
• For automatic dilutions, ensure that ADVIA Centaur CEA Diluent is loaded and set the
system parameters as follows:
Dilution point: ≤ 100 ng/mL (μg/L)
Dilution factor: 5, 10, 50, 100
For detailed information about automatic dilutions, refer to the system operating
instructions or to the online help system.
• Manually dilute the patient samples when patient results exceed the linearity of the assay
using automatic dilution, or when laboratory protocol requires manual dilution.
• Use CEA Diluent to manually dilute patient samples, and then load the diluted sample in
the sample rack, replacing the undiluted sample.
• Ensure that results are mathematically corrected for dilution. If a dilution factor is entered
when scheduling the test, the system automatically calculates the result.
Disposal
Dispose of hazardous and biologically contaminated materials according to the practices of
your institution. Discard all materials in a safe and acceptable manner and in compliance with
all federal, state, and local requirements.
Limitations
NOTE: Do not interpret levels of CEA as absolute evidence of the presence or the absence of
malignant disease. Measurements of CEA should always be used in conjunction with other
diagnostic procedures, including information from the patient’s clinical evaluation.
The concentration of CEA in a given specimen determined with assays from different
manufacturers can vary due to differences in assay methods, calibration, and reagent specificity.
CEA determined with different manufacturers’ assays will vary depending on the method of
standardization and antibody specificity.
WARNING: Do not use the ADVIA Centaur CEA immunoassay as a screening test for diagnosis.
Heterophilic antibodies in human serum can react with reagent immunoglobulins, interfering
with in vitro immunoassays.10 Patients routinely exposed to animals or to animal serum
products can be prone to this interference and anomalous values may be observed. Additional
information may be required for diagnosis.
Serum specimens that are . . . Demonstrate ≤ 5% change in results up to . . .
hemolyzed 500 mg/dL of hemoglobin
lipemic 1000 mg/dL of triglycerides
icteric 20 mg/dL of bilirubin
Expected Results
The expected results for the ACS:180® CEA assay were previously established. Data was
obtained as shown in the following table. Serum samples from healthy subjects and patients
with various malignant diseases were analyzed. The cancer patients included in this study
represented a variety of disease states from active, progressive malignancy to no clinical
evidence of disease. The frequency of positive CEA results was significantly lower in patients
with no evidence of active disease compared to those with active disease.
% Distribution of CEA (ng/mL) (µg/L)
Clinical Condition N 0–2.5 2.6–5.0 5.1–10.0 10.1–20 > 20.0
Healthy Subject
Nonsmokers 225 98.2 1.8 0 0 0
Smokers 150 87.3 8.0 4.7 0 0
Cancerous Diseases
Colorectal 250 37.6 10.8 7.2 6.4 38.0
Lung 158 46.2 15.8 10.8 7.0 20.2
Breast 221 68.3 14.9 7.7 2.3 6.8
Gastric 35 60.0 17.1 8.6 5.7 8.6
Ovarian 35 82.8 11.4 2.9 2.9 0
Pancreatic 43 44.2 20.9 16.3 7.0 11.6
Liver 6 50.0 16.7 0 33.3 0
Lymphoproliferative 15 80.0 20.0 0 0 0
Other† 41 75.0 10.0 10.0 5.0 0
Nonmalignant Diseases
Cirrhosis 53 41.5 13.2 32.1 13.2 0
Other benign liver 5 40.0 20.0 0 40.0 0
Ulcerative colitis 11 90.9 9.1 0 0 0
Benign polyps 23 65.2 21.8 13.0 0 0
Colon and intestinal 15 66.7 20.0 0 13.3 0
Gastrointestinal 21 76.2 14.2 0 4.8 4.8
Breast 53 96.2 0 1.9 0 1.9
Kidney and bladder 12 83.4 8.3 0 8.3 0
Lung 29 69.0 24.1 6.9 0 0
Other‡ 117 86.3 8.5 4.3 0.9 0
† Other cancers includes bladder, prostate, esophagus, head and neck, sarcoma, and kidney.
‡ Other includes benign uterine, cervical and vaginal, benign ovary, and benign male genital as well as other
benign conditions.
These results were confirmed for the ADVIA Centaur CEA assay by analyzing 284 samples in
the range of 0.5 to 78.5 ng/mL (μg/L). Refer to Method Comparison.
As with all diagnostic assays, each laboratory should determine its own reference range(s) for
the diagnostic evaluation of patient results.11
Performance Characteristics
Specificity
The potential interference of NCA (normal cross-reacting antigen) and NCA2 was tested by
adding these antigens to serum pools containing CEA. The level of CEA was then determined.
CEA value without CEA value with
Cross-reactant cross-reactant (ng/mL) (µg/L) cross-reactant (ng/mL) (µg/L)
NCA (500 ng/mL) 2.5 2.5
23.3 21.8
71.2 66.1
NCA2 (100 ng/mL) 2.5 2.5
23.3 22.9
71.2 62.0
NCA and NCA2 showed minimal interference with the recovery of CEA from the serum
samples. Average recovery is greater than 95%.
Method Comparison
For 284 samples in the range of 0.5 to 78.5 ng/mL (μg/L), the relationship between the
ADVIA Centaur CEA assay and the ACS:180 CEA assay is described by the equation:
ADVIA Centaur CEA = 0.97 (ACS:180 CEA) + 0.04 ng/mL
Correlation coefficient (r) = 0.99
Dilution Recovery
Six serum samples in the range of 55.85 to 77.02 ng/mL (μg/L) of CEA were diluted 1:2, 1:4,
1:8, and 1:16 with CEA Diluent and assayed for recovery and parallelism. The recoveries
ranged from 79.2% to 110.9% with a mean of 96.8%.
Observed Expected
Sample Dilution (ng/mL) (µg/L) (ng/mL) (µg/L) Recovery %
1 — 55.85
1:2 28.36 27.93 101.5
1:4 13.67 13.96 97.9
1:8 5.53 6.98 79.2
1:16 3.00 3.49 86.0
Mean 91.2
2 — 57.97
1:2 28.52 29.98 98.4
1:4 13.15 14.49 90.7
1:8 6.42 7.25 88.6
1:16 3.28 3.62 90.4
Mean 92.0
3 — 63.68
1:2 34.43 31.84 108.1
1:4 16.25 15.92 102.1
1:8 8.41 7.96 105.7
1:16 3.81 3.98 95.8
Mean 102.9
4 — 77.02
1:2 40.43 38.51 105.0
1:4 18.42 19.25 95.7
1:8 9.53 9.63 99.0
1:16 4.83 4.81 100.4
Mean 100.0
5 — 70.57
1:2 39.13 35.29 110.9
1:4 18.92 17.64 107.2
1:8 8.27 8.82 93.7
1:16 4.00 4.41 90.7
Mean 100.6
6 — 62.74
1:2 31.23 31.37 99.6
1:4 15.26 15.68 97.3
1:8 7.04 7.84 89.8
1:16 3.55 3.92 90.6
Mean 94.3
Mean 96.8
Spiking Recovery
Varying amounts of CEA were added to seven serum samples with endogenous CEA levels
ranging from 0 to 2.2 ng/mL (µg/L). The amount of CEA that was added varied from 7.0 to
83.0 ng/mL (µg/L). When compared to the expected value, the measured (recovered) values of
CEA averaged 100.8% with a range of 75.5% to 135.8%.
Amount Added Observed
Sample (ng/mL) (µg/L) (ng/mL) (µg/L) Recovery %
1 — 0.5
7.0 6.5 86.6
13.0 10.2 75.5
28.0 28.6 100.5
55.0 53.7 96.8
83.0 80.5 96.5
Mean 91.2
2 — 1.6
7.0 8.3 97.2
13.0 16.7 114.7
28.0 31.3 105.9
55.0 54.5 96.2
83.0 81.0 95.8
Mean 102.0
3 — 0.2
7.0 8.5 118.6
13.0 12.1 91.4
28.0 31.4 111.3
55.0 56.1 101.5
83.0 79.0 94.9
Mean 103.5
4 — 0.8
7.0 7.0 90.0
13.0 11.1 80.7
28.0 32.6 113.2
55.0 56.2 100.8
83.0 82.0 97.9
Mean 96.5
5 — 0.6
7.0 6.9 91.8
13.0 18.4 135.8
28.0 33.1 116.0
55.0 49.9 89.9
83.0 104.4 124.9
Mean 111.7
6 — 0.3
7.0 7.0 95.4
7.0 12.7 95.7
13.0 31.5 111.2
28.0 53.3 96.5
55.0 78.1 93.8
Mean 98.5
7 — 0.7
7.0 7.9 102.2
13.0 12.2 88.7
28.0 31.7 110.6
55.0 60.4 108.5
83.0 83.6 99.8
Mean 102.0
Mean 100.8
Precision
Four samples were assayed 6 times, in each of 12 runs, on 5 systems, (n = 72 for each sample),
over a period of 4 days. The following results were obtained:
Mean (ng/mL) (µg/L) Within-run % CV Run-to-run % CV Total % CV
2.4 3.6 4.1 5.5
14.2 3.3 2.9 4.4
31.5 2.8 4.0 4.8
51.5 2.1 2.7 3.4
Traceability of Standardization
The ADVIA Centaur CEA assay is traceable to an internal standard manufactured using highly
purified material. Assigned values for calibrators are traceable to this standardization.
Evaluating Results
The following is recommended when you observe poor reproducibility of CEA values at low
levels or if you are not satisfied with assay performance:
• Ensure that the assay reagent and calibrator lot numbers and expiration dates match those
entered in the system.
• Ensure that the calibrators, quality control materials, and assay reagents were prepared
according to the recommended procedures.
• Ensure that the recommended sample collection and handling procedures were followed.
• Ensure that the recommended system cleaning procedures were followed.
• Ensure that Type II reagent water was used when operating the system.8
• Visually check the probe and tubing for obstructions, leaks, and deformities such as
pinched or crimped tubing.
• Take further corrective action following established laboratory procedures.
• Calibrate the system using new assay reagents, calibrators, and quality control samples.
• Contact Siemens for technical assistance.
Technical Assistance
For customer support, please contact your local technical support provider or distributor.
www.siemens.com/diagnostics
References
1. Statland BE, Winkel P. Neoplasia. In:Kaplan LA, Pesc AJ, editors. Clinical chemistry; theory, analysis, and
correlation. 2nd ed. St. Louis: CV Mosby, 1989. p.734–5.
2. Sikorska H, Shuster J, Gold P. Clinical applications of carcinoembryonic antigen. Cancer Detection and
Prevention 1988;12:321–55.
3. Lahousen M, Stettner H, Pickel H, et al. The predictive value of a combination of tumor markers in monitoring
patients with ovarian cancer. Cancer 1987;60:2228–32.
4. Go VLW, Zamcheck N. The role of tumor markers in the management of colorectal cancer. Cancer
1982;50:2618–23.
5. McNeely MDD. Gastrointestinal function and digestive disease. In: Kaplan LA, Pesce AJ, editors. Clinical
chemistry; theory, analysis, and correlation. 2nd ed. St. Louis: CV Mosby, 1989. p.411–2.
6. Fletcher RH. Carcinoembryonic Antigen. Annals of Internal Medicine 1986;104:66–73.
7. Minton J, Chevinsky AH. CEA directed second-look surgery for colon and rectal cancer. Annales Chirurgiae et
Gynecologiae 1989;78:32–7.
ADVIA Centaur, ReadyPack, and ACS:180 are trademarks of Siemens Healthcare Diagnostics.
© 2008 Siemens Healthcare Diagnostics Inc. All rights reserved.
US Pats 5,110,932; 5,609,822; 5,788,928
Origin: US