ADVIA Centaur ®

ADVIA Centaur ® XP
ADVIA Centaur ® XPT
Immunoassay Systems

HAV IgM (aHAVM)

Assay for the Detection of IgM Antibodies to Hepatitis A Virus

Current revision and date a Rev. J, 2014-08

Product Name ADVIA Centaur® HAV IgM assay REF 05004126
Systems ADVIA Centaur system
ADVIA Centaur XP system
ADVIA Centaur XPT system
Materials Required but Not ADVIA Centaur HAV IgM Quality Control Material REF 05004800
Provided ADVIA Centaur Multi-Diluent 2 REF 07948423
(110314)
ADVIA Centaur Wash 1 (2 x 1500 mL) REF 01137199
ADVIA Centaur Wash 1 (2 x 2500 mL) REF 03773025
Specimen Types Serum, EDTA plasma, Lithium or sodium heparinized plasma
Assay Range 0.02–7.00 S/CO
Reagent Storage 2–8°C
Reagent On-System Stability 41 days
a In Rev. B or later, a vertical bar in the margin indicates a technical update to the previous version.

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.

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Summary and Explanation

The ADVIA Centaur HAV IgM assay is an antibody capture microparticle chemiluminometric
immunoassay used for the detection of IgM antibody to hepatitis A virus in human serum or
plasma.
Hepatitis A is caused by infection with the hepatitis A virus. HAV is a 27 nanometer
single-stranded, nonenveloped, RNA virus that is classified as a picornavirus. Transmission of
hepatitis A is via the fecal-oral route and infection occurs mainly due to contaminated food or
poor sanitary conditions.1,2
Hepatitis A virus replicates in the liver. The virus is excreted in the bile and shed in the stool.
Only one serotype has been observed among HAV isolates collected from various parts of the
world. The average incubation period for HAV infection is 30 days with a range of 15 to
40 days. Chronic infection has not been reported to occur following HAV infection. Symptoms
last approximately 2 weeks and include hepatomegaly, jaundice, dark urine, fatigue, and
gastrointestinal distress such as anorexia, nausea, vomiting, and abdominal pain. At the onset
of symptoms resulting from HAV infection, antibody to HAV is detectable. The early antibody
response is largely comprised of the IgM antibody subclass. Anti-HAV IgM is detectable usually
for 3 to 6 months after the onset of illness, whereas anti-HAV IgG can persist indefinitely.
Because of the transient production of anti-HAV IgM, its presence in sera indicates ongoing or
recent infection and is the most useful serological marker for diagnosing acute HAV
infection.1–4
Since symptomatic hepatitis A viral infections can be clinically indistinguishable from
hepatitis B or C viral infections, serological testing is important for proper diagnosis.

Principles of the Procedure

The ADVIA Centaur HAV IgM assay is an IgM capture immunoassay using a 2-pass format. In
the first pass the sample is diluted using Multi-Diluent 2. After sample dilution biotinylated
anti-human IgM monoclonal antibody is added to the cuvette binding IgM from the diluted
patient sample. The IgM complex is then captured by the addition of streptavidin coated
magnetic latex particles (MLP). The IgM-MLP is washed and resuspended.
In the second step pass the anti-HAV IgM captured on the Solid Phase is detected by the
sequential addition of HAV antigen and acridinium ester-labeled mouse anti-HAV antibody.

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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

ADVIA Centaur 1500 mL/pack 2–25°C Unopened: Stable until the

Wash 1a phosphate-buffered saline with sodium azide expiration date on the pack
(< 0.1%) and surfactant On-system: 1 month

ADVIA Centaur 2500 mL/pack 2–25°C Unopened: Stable until the

Wash 1a phosphate-buffered saline with sodium azide expiration date on the pack
(< 0.1%) and surfactant On-system: 1 month

ADVIA Centaur 10.0 mL/reagent pack 2–8°C Unopened: Until the

ReadyPack goat serum with sodium azide (< 0.1%) and expiration date on the pack
ancillary reagent preservatives On-system: 28 consecutive
pack; days after accessing the
Multi-Diluent 2a ancillary reagent pack

a See Materials Required but Not Provided.

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Warnings and Precautions

Safety data sheets (MSDS/SDS) available on www.siemens.com/diagnostics.

CAUTION POTENTIAL BIOHAZARD

Some components of this product contain human source material. No known test method can offer
complete assurance that products derived from human blood will not transmit infectious agents. All
products manufactured using human source material should be handled as potentially infectious. Handle
this product according to established good laboratory practices and universal precautions.5–7
The negative control has been assayed by FDA-approved methods and found nonreactive for hepatitis B
surface antigen (HBsAg), antibody to hepatitis C (HCV), and antibody to HIV-1/2. The positive control and
calibrators contain human plasma that is reactive for anti-HAV IgM. The units were inactivated using a
BPL-UV inactivation procedure. The Ancillary Well Reagent contains HAV virus inactivated with formalin.
All products manufactured using human source material should be handled as potentially infectious.

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

Dispose of hazardous or biologically contaminated materials according to the practices of your

institution. Discard all materials in a safe and acceptable manner and in compliance with
prevailing regulatory requirements.
For in vitro diagnostic use.

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.

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Storing and Stability

Store the reagents upright at 2–8°C.
Protect reagent packs from all heat and light sources. Reagent packs loaded on the system are
protected from light. Store unused reagent packs at 2–8°C away from heat and light sources.
All reagents are stable at 2–8°C until the expiration date on the packaging.

Specimen Collection and Handling

Serum, EDTA plasma, lithium or sodium heparinized plasma are the recommended sample
types for this assay. Do not use specimens with obvious microbial contamination. 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 following general recommendations for handling and storing blood samples are furnished
by the Clinical and Laboratory Standards Institute (CLSI),8 and augmented with additional
sample handling studies using the ADVIA Centaur HAV IgM assay:
• Handle all samples as if capable of transmitting disease.
• Samples are processed by centrifugation, typically followed by physical separation of the
serum or plasma from the red cells. The centrifugation step may occur up to 24 hours post
draw. When testing 10 samples where the centrifugation step was varied up to 24 hours
post draw, no clinically significant differences were observed.
• Test samples as soon as possible after collecting. Store samples at 2–8°C if not tested
immediately.
• Store samples stoppered and upright at all times at 2–8°C up to 7 days.
• Store primary tube samples at 2–8°C up to 7 days. Keep samples stoppered and upright at
all times. Primary tube samples include serum stored on the clot, plasma stored on packed
red cells, and samples processed and stored in gel barrier blood collection tubes. When
10 samples in these primary tubes were tested up to 7 days, no clinically significant
differences were observed.
• Freeze samples, devoid of red blood cells, at or below -20°C for longer storage. Samples
may be stored at or below -20°C for up to 180 days. Do not store in a frost-free freezer.
When 10 samples were subject to 4 freeze/thaw cycles, no clinically significant differences
were observed. Thoroughly mix thawed samples and centrifuge before using.
• Package and label samples for shipment in compliance with applicable federal and
international regulations covering the transport of clinical samples and etiological agents.
Samples maintained at room temperature up to 2 days or refrigerated up to 7 days
demonstrated no qualitative differences; however, good laboratory practice indicates that
samples should be stored refrigerated. Store samples stoppered and upright at 2–8°C upon
arrival. If shipment is expected to exceed 7 days, ship specimens frozen.
The purpose of handling and storage information is to provide guidance to users. It is the
responsibility of the individual laboratory to use all available references and/or its own studies
when establishing alternate stability criteria to meet specific needs.

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Procedure
Materials Provided
The following materials are provided:

REF Contents Number of Tests

05004126 1 ReadyPack primary reagent pack containing ADVIA Centaur 100
HAV IgM Lite Reagent, Solid Phase, and Ancillary Well Reagent
1 Ancillary pack containing ADVIA Centaur HAV IgM Ancillary
Reagent
ADVIA CentaurHAV IgM Master Curve card
1 vial ADVIA Centaur HAV IgM low calibrator
1 vial ADVIA Centaur HAV IgM high calibrator
ADVIA Centaur HAV IgM Calibrator Assigned Value card

Materials Required but Not Provided

The following materials are required to perform this assay, but are not 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.

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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.

Preparing the System

Ensure that the system has sufficient primary and ancillary reagent packs. For detailed
information about preparing the system, refer to the system operating instructions.
Load the ReadyPack primary reagent packs in the primary reagent compartment using the
arrows on the packs as a placement guide. The system automatically mixes the primary reagent
packs to maintain homogeneous suspension of the reagents. Load the Multi-Diluent 2 ancillary
reagent pack in the ancillary reagent entry. For detailed information about loading reagents,
refer to the system operating instructions.

Preparing the Samples

This assay requires 20 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 the system operating instructions.
Before placing samples on the system, ensure that samples have the following characteristics:
• Samples are free of fibrin or other particulate matter. Remove particulates by
centrifugation.
• Samples are free of bubbles or foam.

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.

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Perform the calibration procedure using the following steps:

Note This procedure uses calibrator volumes sufficient to measure each calibrator in
duplicate.
1. Schedule the calibrators to the worklist.
2. Label two sample cups with calibrator barcode labels: one for the low and another for the
high.
3. Gently mix the Low and High Calibrators and dispense at least 4 to 5 drops into the
appropriate sample cups.
Note Each drop from the calibrator 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 calibrator remaining in the sample cups after 8 hours. Do not refill
sample cups when the contents are depleted; if required, dispense fresh calibrators.

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.

Using Barcode Labels

Note Calibrator barcode labels are lot-number specific. Do not use barcode labels from one
lot of calibrators with any other lot of calibrators.
Use the ADVIA Centaur HAV IgM Calibrator barcode labels to identify the Low and High
Calibrator sample cups when performing the ADVIA Centaur HAV IgM assay. Place the
barcode label on the sample cup so that the readable characters on the side of the label are
vertical on the sample cup.

Performing Master Curve Calibration

The ADVIA Centaur HAV IgM assay requires a Master Curve calibration when using a new lot
number of Lite Reagent, Solid Phase, and Ancillary Well Reagent. For each new lot number of
Lite Reagent, Solid Phase, and Ancillary Well Reagent, use the barcode reader or keyboard to
enter the Master Curve values on the system. The Master Curve card contains the Master Curve
values. For detailed information about entering calibration values, refer to the system
operating instructions.

Performing Quality Control

Follow government regulations or accreditation requirements for quality control frequency.
For quality control of the ADVIA Centaur HAV IgM assay, use ADVIA Centaur HAV IgM quality
control material. Refer to the Expected Value card for the suggested expected values specific
for the lot number of the positive and negative controls.
For detailed information about entering quality control values, refer to the system operating
instructions.

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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.

Using Barcode Labels

Note Control barcode labels are lot-number specific. Do not use barcode labels from one lot
of controls with any other lot of controls.
Use the ADVIA Centaur HAV IgM quality control barcode labels to identify the positive and
negative sample cups when performing the ADVIA Centaur HAV IgM assay. Place the barcode
label on the sample cup so that the readable characters on the side of the label are vertical on
the sample cup.

Taking Corrective Action

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.

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.

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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.

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Clinical Sensitivity and Specificity

Clinical Specificity
A population of 515 hospitalized patients and 103 recovered HAV patients was tested using the
ADVIA Centaur HAV IgM assay and a commercially available automated anti-HAV IgM EIA. The 1
sample initially ADVIA Centaur HAV IgM equivocal and nonreactive using the comparative anti-
HAV IgM assay resolved as concordant reactive following consensus method testing. The
relative specificity of the ADVIA Centaur HAV IgM was 100% (616/616).
Relative Specificity

Comparative HAV IgM Assay

ADVIA Centaur HAV IgM Assay Reactive Equivocal Nonreactive Total

Reactive 1 0 0 1
Equivocal 0 0 1 1
Nonreactive 0 0 616 616

Total 1 0 617 618

Relative Specificity = 100% (616/616), 95% CI (Confidence Interval) = 99.40–100%

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

Comparative Anti-HAV IgM Assay

ADVIA Centaur HAV IgM Assay Reactive Equivocal Nonreactive Total

Reactive 93 3 0 96
Equivocal 1 0 0 1
Nonreactive 0 3 1 4

Total 94 6 1 101

Resolved Clinical Sensitivity

Further analysis of the 7 samples with equivocal results (1 ADVIA Centaur equivocal/
comparative assay reactive, 3 ADVIA Centaur nonreactive/comparative assay equivocal, and
3 ADVIA Centaur reactive/comparative assay equivocal) was performed using an additional
commercially available assay for anti-HAV IgM.
The sample that was equivocal using the ADVIA Centaur HAV IgM assay and reactive using the
comparative assay, became ADVIA Centaur equivocal/consensus method reactive.

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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

Consensus HAV IgM Assay Results

ADVIA Centaur HAV IgM Assay Reactive Equivocal Nonreactive Total

Reactive 96 0 0 96
Equivocal 2 0 0 2
Nonreactive 0 2 1 3

Total 98 2 1 101

Resolved Sensitivity = 100% (96/96), 95% CI (Confidence Interval) = 96.23–100%

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

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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.

Within-run Between run Total

Mean
Sample S/CO SD CV (%) SD CV (%) SD CV (%)
Negative Control 0.13 0.00 N/Aa 0.01 N/A 0.01 N/A
Positive Control 1.77 0.06 3.7 0.12 6.5 0.13 7.5
K2 EDTA 1 0.19 0.01 N/A 0.00 N/A 0.01 N/A
K2 EDTA 2 0.67 0.03 3.8 0.03 4.0 0.04 6.2
K2 EDTA 3 1.24 0.05 4.2 0.06 4.5 0.08 6.7
K2 EDTA 4 1.69 0.07 4.3 0.07 4.3 0.12 6.8
K2 EDTA 5 2.31 0.10 4.5 0.14 6.1 0.21 9.0
Lithium heparin 1 0.16 0.00 N/A 0.00 N/A 0.01 N/A
Lithium heparin 2 0.69 0.03 4.4 0.02 3.3 0.04 5.7
Lithium heparin 3 1.32 0.08 6.0 0.06 4.6 0.10 7.6
Lithium heparin 4 1.81 0.11 5.9 0.11 5.9 0.15 8.3
Lithium heparin 5 2.50 0.19 7.6 0.10 3.8 0.21 8.5
Sodium heparin 1 0.29 0.01 N/A 0.00 N/A 0.01 N/A
Sodium heparin 2 0.72 0.04 5.6 0.02 2.3 0.05 6.5
Sodium heparin 3 1.46 0.11 7.4 0.05 3.6 0.14 9.7
Sodium heparin 4 1.98 0.14 7.0 0.12 6.3 0.20 9.9
Sodium heparin 5 2.71 0.24 8.7 0.13 4.8 0.28 10.3
Serum 1 0.17 0.01 N/A 0.01 N/A 0.01 N/A
Serum 2 0.69 0.03 4.1 0.02 3.4 0.04 6.2
Serum 3 1.22 0.06 4.9 0.04 3.3 0.08 6.4
Serum 4 1.75 0.09 4.9 0.07 4.3 0.12 6.8
Serum 5 2.52 0.12 4.9 0.13 5.2 0.18 7.3
a N/A = Not Applicable

Interferences

Serum specimens that are . . . Demonstrate ≤ 10% change in results up to . . .

hemolyzed 500 mg/dL of hemoglobin
lipemic 3000 mg/dL of triglycerides
icteric 60 mg/dL of conjugated bilirubin
icteric 40 mg/dL of unconjugated bilirubin
proteinemic 12 g/dL of protein
proteinemic 3.5 g/dL of protein

Interference testing was determined according to CLSI Document EP7-A2.13

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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.

ADVIA Centaur HAV IgM Results

Number
Clinical Category Tested Nonreactive Equivocal Reactive
Hepatitis B Infection (HBV) 2 2 0 0
Hepatitis C Infection (HCV) 11 11 0 0
Non-viral Liver Disease 8 8 0 0
Epstein-Barr Virus (EBV) IgG 10 9 1 0
Epstein-Barr Virus (EBV) IgM 10 10 0 0
Herpes Simplex Virus (HSV) IgG 10 10 0 0
Herpes Simplex Virus (HSV) IgM 10 10 0 0
Syphilis IgG 10 10 0 0
Syphilis IgM 10 10 0 0
Human Immunodeficiency Virus 10 10 0 0
(HIV 1/2)
Varicella Zoster (VZV) IgG 10 10 0 0
Cytomegalovirus (CMV) IgG 2 2 0 0
Cytomegalovirus (CMV) IgM 3 3 0 0
Toxoplasma IgG 10 10 0 0
Toxoplasma IgM 9 9 0 0
Rubella IgG 10 10 0 0
Alcoholic Hepatitis 2 2 0 0
Multiparity 10 10 0 0
Flu Vaccine Recipient 6 6 0 0
Rheumatoid Arthritis (RF) 10 10 0 0
Anti-Nuclear Antibody (ANA) 10 10 0 0
Systemic Lupus Erythematosus (SLE) 10 10 0 0
Human Anti-Mouse Antibodies (HAMA) 9 9 0 0

Total Samples Tested 192 191 1 0

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

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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.

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Definition of Symbols
The following symbols may appear on the product labeling:

Symbol Definition Symbol Definition

In vitro diagnostic medical device Catalog number

Authorized Representative in
Legal manufacturer
the European Community

CE Mark with identification number

CE Mark
of notified body

Consult instructions for use Biological risk

Do not freeze (> 0°C) Temperature limitation

Lower limit of temperature Upper limit of temperature

Keep away from sunlight and heat Up

Use by Contains sufficient for (n) tests

Shake the reagent pack vigorously.

Refer to Preparing Reagents in the
Batch code assay-specific ADVIA Centaur
product instructions for detailed
information.

YYYY-MM-DD Date format (year-month-day) Rev. Revision

Variable hexadecimal number that

ensures the Master Curve and
Master Curve Definition
Calibrator definition values entered
are valid.

Lot Details Green dot

Recycle Printed with soy ink

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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

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