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ADVIA Centaur ® XP
ADVIA Centaur ® XPT
Immunoassay Systems
Intended Use
The ADVIA Centaur® HAV IgM (aHAVM) assay is an in vitro diagnostic immunoassay for the
qualitative determination of IgM response to the hepatitis A virus (HAV) in human serum or
plasma (EDTA, lithium or sodium heparinized) using the ADVIA Centaur, ADVIA Centaur XP, and
ADVIA Centaur XPT systems. This assay is intended for use as an aid in the diagnosis of acute or
recent infection (usually 6 months or less) with hepatitis A virus.
Reagents
Reagent Description Storage Reagent Stability
ADVIA Centaur 5.0 mL/reagent pack 2–8°C Unopened: Stable until the
HAV IgM anti-HAV monoclonal antibody (F(ab)2 expiration date on the
ReadyPack® fragment; ~0.3 µg/mL) labeled with acridinium carton
primary reagent ester in buffer with bovine serum albumin, On-system: 41 days
pack; surfactant, sodium azide (< 0.1%), and
Lite Reagent preservatives
ADVIA Centaur 15.0 mL/reagent pack 2–8°C Unopened: Stable until the
HAV IgM streptavidin coated paramagnetic expiration date on the
ReadyPack microparticles in buffer with bovine serum carton
primary reagent albumin, surfactant, sodium azide (< 0.1%), On-system: 41 days
pack; and preservatives
Solid Phase
Reagent
ADVIA Centaur 5.0 mL/reagent pack 2–8°C Unopened: Stable until the
HAV IgM inactivated hepatitis A virus (< 0.1 µg/mL) in expiration date on the
ReadyPack buffer with bovine serum albumin, surfactant, carton
primary reagent sodium azide (< 0.1%), and preservatives On-system: 41 days
pack;
Ancillary Well
Reagent
ADVIA Centaur 25.0 mL/reagent pack 2–8°C Unopened: Stable until the
HAV IgM biotinylated monoclonal mouse anti-human expiration date on the
Readypack IgM (~0.500 µg/mL) in buffer with bovine carton
ancillary reagent serum albumin, mouse IgG, surfactant, sodium On-system: 41 days
pack; azide (< 0.1%), and preservatives
Ancillary
Reagent
ADVIA Centaur 2.0 mL/vial 2–8°C Unopened: Stable until the
HAV IgM processed human plasma positive for IgM expiration date on the vial
calibrator antibodies to HAV with preservatives On-system: 8 hours
ADVIA Centaur 7.0 mL/vial 2–8°C Unopened: Stable until the
HAV IgM quality processed human plasma negative and expiration date on the vial
control materiala positive for IgM antibodies to HAV, with On-system: 8 hours
preservatives
CAUTION
This device contains material of animal origin and should be handled as a potential carrier and
transmitter of disease.
Contains sodium azide as a preservative. Sodium azide can react with copper or lead plumbing
to form explosive metal azides. On disposal, flush reagents with a large volume of water to
prevent buildup of azides. Disposal into drain systems must be in compliance with prevailing
regulatory requirements.
H412 Harmful to aquatic life with long lasting effects.
P273, P501 Avoid release to the environment. Dispose of contents and container in accordance
with all local, regional, and national regulations.
Contains: Microprotect; ADVIA Centaur HAV IgM calibrator
Preparing Reagents
All reagents are liquid and ready to use.
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 the system operating
instructions.
Note
The Ancillary Reagent provided in this kit is matched to the Lite Reagent, Solid Phase, and
Ancillary Well Reagent. Do not mix Ancillary Reagent lots with different lots of Lite Reagent,
Solid Phase, and Ancillary Well Reagent.
The Ancillary Reagent pack contains more volume that required to perform 100 tests. Since
the Ancillary Reagent is matched to the Lite Reagent, Solid Phase, and Ancillary Well Reagent in
the ReadyPack primary reagent pack, discard the Ancillary Reagent pack when the ReadyPack
primary reagent pack is discarded. Do not use beyond the on-system stability.
Note
• Discard reagent packs at the end of the on-system stability interval.
• Do not use reagents beyond the expiration date.
Procedure
Materials Provided
The following materials are provided:
Item Description
REF 05004800 ADVIA Centaur HAV IgM quality control 2 x 7.0 mL negative control
material 2 x 7.0 mL positive control
Expected Value card
REF 07948423 ADVIA Centaur Multi-Diluent 2a 2 ReadyPack ancillary reagent packs
(110314) containing 10 mL/pack
REF 01137199 ADVIA Centaur Wash 1 2 x 1500 mL/pack
(112351)
REF 03773025 ADVIA Centaur Wash 1b 2 x 2500 mL/pack
a A minimum of two ADVIA Centaur Multi-Diluent 2 ancillary reagent packs are required for each
ADVIA Centaur HAV IgM primary reagent pack (100 tests).
b For use with systems with 2500 mL capacity.
Assay Procedure
For detailed instructions on performing the procedure, refer to the system operating
instructions.
The system automatically performs the following steps:
1. Dispenses 20 µL of sample and 180 µL of Multi-Diluent 2 into a cuvette.
2. Aspirates 60 µL of diluted sample and dispenses it into a cuvette.
3. Dispenses 150 µL of Ancillary Reagent and incubates for 6 minutes at 37°C.
4. Dispenses 150 µL of Solid Phase and incubates for 18 minutes at 37°C.
5. Separates the Solid Phase from the mixture and aspirates the unbound reagent.
6. Washes the cuvette with Wash 1.
7. Resuspends the particles in 250 µL of Wash 1 and incubates for 6.75 minutes at 37°C.
8. Dispenses 50 µL each of Ancillary Well Reagent and Lite Reagent and incubates for
18 minutes at 37°C.
9. Separates the Solid Phase from the mixture and aspirates the unbound reagent.
10. Washes the cuvette with Wash 1.
11. Dispenses 300 µL each of Acid Reagent and Base Reagent to initiate the chemiluminescent
reaction.
12. Reports results according to the selected option, as described in the system operating
instructions.
A direct relationship exists between the amount of anti-HAV IgM activity present in the patient
sample and the amount of relative light units (RLUs) detected by the system. A result of
reactive or nonreactive is determined according to the Signal-to-Cutoff (S/CO) Value
established with the calibrators. Refer to Interpretation of Results for a description of the Cutoff
Value calculation.
On-System Stability
The ADVIA Centaur aHAVM assay reagents are stable unopened until the expiration date on the
carton or onboard the system for 41 days.
Performing Calibration
For calibration of the ADVIA Centaur HAV IgM assay, use ADVIA Centaur HAV IgM Calibrators
provided with each kit. The calibrators provided in this kit are matched to the ReadyPack
primary reagent pack.
Note The Low and High Calibrators provided in this kit are matched to the ReadyPack primary
reagent pack. Do not mix calibrator lots with different lots of reagent packs.
Each lot of calibrators contains a Calibrator Assigned Value card to facilitate entering the
calibration values on the system. Enter the values using the barcode scanner or the keyboard.
For detailed information about entering calibrator values, refer to the system operating
instructions.
Calibration Frequency
Calibrate the assay at the end of the 28-day calibration interval.
Additionally, the ADVIA Centaur HAV IgM assay requires a two-point calibration:
• When changing lot numbers of primary reagent packs.
• When replacing system components.
• When quality control results are repeatedly out of range.
To monitor system performance and chart trends, as a minimum requirement, quality control
samples should be assayed on each workshift 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.
Perform the quality control procedure using the following steps:
Note This procedure uses control volumes sufficient to measure each control in duplicate.
1. Schedule the quality control samples to the worklist.
2. Label two sample cups with quality control barcode labels: one for the positive, and
another for the negative.
3. Gently mix the quality control materials and dispense at least 4 to 5 drops into the
appropriate sample cups.
Note Each drop from the control vial is approximately 50 µL.
4. Load the sample cups in a rack.
5. Place the rack in the sample entry queue.
6. Ensure that the assay reagents are loaded.
7. Start the entry queue, if required.
Note Dispose of any quality control materials remaining in the sample cups after 8 hours. Do
not refill sample cups when the contents are depleted; if required, dispense fresh quality
control materials.
Results
Calculation of Results
For detailed information about how the system calculates results, refer to the system operating
instructions.
The system reports HAV IgM results in S/CO Values and as reactive, equivocal, or nonreactive.
Interpretation of Results
• Samples with a calculated value of less than 0.80 S/CO Value are considered nonreactive
for IgM antibodies to hepatitis A virus.
• Samples with a calculated value greater than or equal to 0.80 S/CO Value and less than
1.20 S/CO Value are considered equivocal and must be repeated. It is recommended that
the test be repeated in duplicate and the results be reported based on the repeat results. If
the results are still equivocal after repeat testing, obtain a new specimen and retest using
the ADVIA Centaur HAV IgM assay.
• Samples with a calculated value greater than or equal to 1.20 S/CO Value are considered
reactive for IgM antibodies to hepatitis A virus.
• The cutoff for the ADVIA Centaur HAV IgM assay was verified based on results of Receiver-
Operator characteristics (ROC) Curve9 and clinical agreement generated from the clinical
studies.
• Sample results are invalid and must be repeated if the controls are out of range.
Limitations
The following information pertains to limitations of the assay:
• The ADVIA Centaur HAV IgM assay is limited to the detection of IgM antibodies to hepatitis
A virus in human serum or plasma (EDTA plasma, lithium or sodium heparinized plasma).
• The ADVIA Centaur HAV IgM assay can be used to determine if a patient has or recently had
an acute or asymptomatic hepatitis A infection. This test does not measure anti-HAV IgG
and therefore cannot be used to determine a patient’s immune status to hepatitis A.
• The performance of the ADVIA Centaur HAV IgM assay has not been established with cord
blood, neonatal specimens, cadaver specimens, heat-inactivated specimens, or body fluids
other than serum or plasma, such as saliva, urine, amniotic, or pleural fluids.
• The performance of the assay has not been established for populations of
immunocompromised or immunosuppressed patients.
• Do not use specimens with obvious microbial contamination.
• 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.
Expected Values
In a population of 515 hospitalized/clinical samples, 99.6% (513/515) were nonreactive using
the ADVIA Centaur HAV IgM assay. Of 98 anti-HAV IgM consensus reactive samples, 96 were
reactive and 2 were equivocal using the ADVIA Centaur HAV IgM assay.
As with all in vitro diagnostic assays, each laboratory should determine its own reference
range(s) for the diagnostic evaluation of patient results.11
Performance Characteristics
The performance of the ADVIA Centaur HAV IgM assay was determined by testing a total of
719 samples at 2 sites. The ADVIA Centaur HAV IgM results were compared to test results using
a commercially available automated anti-HAV IgM EIA. The samples included the following
populations: 101 acute HAV patients, 515 hospitalized patients, and 103 recovered HAV
patients. Further evaluation was performed with the discordant and equivocal samples using
another commercially available assay for anti-HAV IgM.
Clinical Sensitivity
A population of 101 acute HAV patient samples was tested using the ADVIA Centaur HAV IgM
assay and a commercially available anti-HAV IgM EIA. 94 of these HAV patient samples were
found reactive for anti-HAV IgM using the comparative assay with the same intended use. Of
these reactive samples, 93 were reactive and 1 was equivocal using the ADVIA Centaur HAV
IgM assay. 6 samples were comparative assay equivocal and 1 sample was nonreactive using
both assays. The initial relative sensitivity was 100%.
Note Samples giving equivocal results were not included in the calculation of relative
sensitivity and relative specificity.
Initial Sensitivity
Total 94 6 1 101
Of the 3 samples that were nonreactive using the ADVIA Centaur HAV IgM assay and equivocal
using the comparative assay, 1 became ADVIA Centaur nonreactive/consensus method
equivocal, 1 became ADVIA Centaur equivocal/consensus method reactive, and 1 remained
indeterminate (ADVIA Centaur nonreactive/comparative assay equivocal/consensus method
reactive).
Of the 3 samples that were reactive using the ADVIA Centaur HAV IgM assay and equivocal
using the comparative assay, all 3 became consensus method reactive.
The consensus results are shown in the following table:
Resolved Clinical Sensitivity
Total 98 2 1 101
Seroconversion Panels
The clinical sensitivity of the ADVIA Centaur HAV IgM assay has been set so that a reactive
result implies an acute or recent HAV infection. Anti-HAV IgM antibody is usually present for 6
months or less after the onset of illness. This is illustrated by the following serial bleed profile
from a hepatitis A patient.
Hepatitis A Patient Serial Bleed Profile
Precision
Precision was evaluated according to the CLSI protocol EP5-A.12 A five member panel and
controls were assayed in three replicates twice a day for 20 days. The following results were
obtained using one reagent lot and a stored calibration curve.
Interferences
Cross-Reactivity
The ADVIA Centaur HAV IgM assay was evaluated for potential cross-reactivity with viral
antibodies and disease state specimens. The nonreactive anti-HAV IgM status of each specimen
was verified using an anti-HAV IgM comparative assay. The following results were obtained
using the ADVIA Centaur HAV IgM assay.
Standardization
The ADVIA Centaur HAV IgM assay standardization is based upon relative clinical agreement
with commercially available anti-HAV IgM assays. The ADVIA Centaur HAV IgM assay cutoff has
been set to detect acute or recent (usually six months or less) hepatitis A infection. Refer to
Performance Characteristics. Assigned values for calibrators and controls are traceable to this
standardization.
Technical Assistance
For customer support, please contact your local technical support provider or distributor.
www.siemens.com/diagnostics
References
1. Hollinger FB, Ticehurst JR. Hepatitis A virus. In: Fields BN, Knipe DM, Howley PM.
Fields Virology, 3rd ed. Philadelphia, PA: Lippincott-Raven Publishers; 1996:735–782.
2. Stapleton JT. Hepatitis A virus: biology, pathogenesis, epidemiology, clinical description,
and diagnosis. In: Specter S. Viral Hepatitis: Diagnosis, Therapy, and Prevention.
Totowa, NJ: Humana Press Inc.; 1999:7–33.
3. Cuthbert JA. Hepatitis A: old and new. Clin Microbiol Rev. 2001;14(1):38–58.
4. Bower WA, Nainan OV, Han X, Margolis HS. Duration of viremia in hepatitis A virus
infection. J Infect Dis. 2000;182:12–17.
5. Centers for Disease Control. Update: Universal precautions for prevention of transmission
of human immunodeficiency virus, hepatitis B virus and other bloodborne pathogens in
healthcare settings. MMWR. 1988;37:377–82, 387–8.
6. Clinical and Laboratory Standards Institute (formerly NCCLS). Protection of Laboratory
Workers From Occupationally Acquired Infections; Approved Guideline - Third Edition.
Wayne, PA: Clinical and Laboratory Standards Institute; 2005. CLSI Document M29-A3.
7. Federal Occupational Safety and Health Administration, Bloodborne Pathogens Standard,
29 CFR 1910.1030.
8. Clinical and Laboratory Standards Institute (formerly NCCLS). Procedures for the Handling
and Processing of Blood Specimens; Approved Guideline - Third Edition. Wayne, PA:
Clinical and Laboratory Standards Institute; 2004. NCCLS Document H18-A3.
9. Clinical and Laboratory Standards Institute (formerly NCCLS). Assessment of the Clinical
Accuracy of Laboratory Tests Using Receiver Operating Characteristic (ROC) Plots;
Approved Guideline. Wayne, PA: Clinical and Laboratory Standards Institute; 1995.
NCCLS Document GP10-A.
10. Boscato LM, Stuart MC. Heterophilic antibodies: a problem for all immunoassays.
Clin Chem. 1988;34:27–33.
11. Clinical and Laboratory Standards Institute (formerly NCCLS). How to Define and
Determine Reference Intervals in the Clinical Laboratory; Approved Guideline -
Second Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2000.
NCCLS Document C28-A2.
12. Clinical and Laboratory Standards Institute (formerly NCCLS). Evaluation of Precision
Performance of Quantitative Measurement Methods; Approved Guideline - Second Edition.
Wayne, PA: Clinical and Laboratory Standards Institute; 2004. NCCLS Document EP5-A2.
13. Clinical and Laboratory Standards Institute (formerly NCCLS). Interference Testing in
Clinical Chemistry; Approved Guideline - Second Edition. Wayne, PA:
Clinical and Laboratory Standards Institute; 2005. CLSI Document EP7-A2.
Definition of Symbols
The following symbols may appear on the product labeling:
Authorized Representative in
Legal manufacturer
the European Community
Trademarks
ADVIA Centaur and ReadyPack are trademarks of Siemens Healthcare Diagnostics.
© 2014 Siemens Healthcare Diagnostics. All rights reserved.
US Pats 5,609,822; 5,788,928