en

Rubella IgG
08P46
Rubella IgG Reagent Kit G71305R04
B8P460
Read Highlighted Changes: Revised January 2020.

08P4622
08P4632
Instructions must be carefully followed. Reliability of assay results ll
BIOLOGICAL PRINCIPLES OF THE PROCEDURE
cannot be guaranteed if there are any deviations from these This assay is a two-step immunoassay for the quantitative
instructions. determination and qualitative detection of IgG antibodies to
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NAME rubella virus in human serum and plasma using chemiluminescent
Alinity i Rubella IgG Reagent Kit microparticle immunoassay (CMIA) technology.
Sample, assay diluent, and partially purified rubella virus coated
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INTENDED USE paramagnetic microparticles are combined and incubated. The
The Alinity i Rubella IgG assay is a chemiluminescent microparticle IgG antibodies to rubella present in the sample binds to the rubella
immunoassay (CMIA) used for the quantitative determination and virus coated microparticles. The mixture is washed. Anti-human IgG
qualitative detection of IgG antibodies to rubella virus in human acridinium-labeled conjugate is added to create a reaction mixture
serum and plasma on the Alinity i analyzer. and incubated. Following a wash cycle, Pre-Trigger and Trigger
The Alinity i Rubella IgG assay is to be used as an aid in the Solutions are added.
determination of immune status to rubella. The resulting chemiluminescent reaction is measured as relative light
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SUMMARY AND EXPLANATION OF THE TEST units (RLUs). There is a direct relationship between the amount of
IgG antibodies to rubella in the sample and the RLUs detected by the
Primary postnatal rubella virus infection is typically a mild self-limiting
disease characterized by a maculopapular rash, fever, malaise system optics.
and lymphadenopathy.1 In contrast to postnatal infections, primary For additional information on system and assay technology, refer to
prenatal infections may have devastating effects. In utero infections the Alinity ci-series Operations Manual, Section 3.
may severely damage the fetus, particularly if occurring during the ll
REAGENTS
first four months of gestation. The congenitally infected infant may
exhibit one or more of a variety of defects collectively known as
Kit Contents
the congenital rubella syndrome (CRS). Among these are low birth Alinity i Rubella IgG Reagent Kit 08P46
weight, cataracts, deafness, congenital heart disease, and mental NOTE: Some kit sizes are not available in all countries. Please
retardation.1 The World Health Organization (WHO) conducted a contact your local distributor.
worldwide survey on rubella, CRS, and rubella vaccine in 1995 and Volumes (mL) listed in the table below indicate the volume per
1996. They reported an incidence of CRS of 60 to 220 cases with cartridge.
100 000 live births during epidemics in developing countries, a rate
08P4622 08P4632
similar to those of industrialized countries before vaccination.2
Tests per cartridge 100 500
Both naturally acquired and vaccine induced immunity to rubella virus
Number of cartridges
associated with antibody persistence have been shown to provide 2 2
per kit
protection from clinical rubella upon reinfection.3-6 The widespread
Tests per kit 200 1000
use of highly effective and safe vaccines dramatically reduced the
incidence of rubella and CRS in the United States. In spite of this 6.6 mL 27.0 mL
reduction, rubella outbreaks continue to occur. The number of cases 6.1 mL 26.5 mL
of rubella reported annually to the WHO Regional Office for Europe 10.4 mL 47.1 mL
has remained fairly stable over the past decade, with 304 320
cases reported during 2003. These occurrences indicate a need for Partially purified rubella virus coated microparticles
continued serological surveillance to identify susceptible individuals in TRIS buffer with surfactant. Preservatives: sodium azide and
and reduce the potential risk to CRS.2 ProClin 950.
Specific antibodies correlate with immunity, but it has not been Anti-human IgG (mouse, monoclonal) acridinium-
possible to identify a specific type and level of antibodies which are labeled conjugate in MES buffer with surfactant and protein
invariably correlated with protection. Traditionally rubella IgG antibody (bovine) stabilizer. Minimum concentration: 16 ng/mL. Preservatives:
levels >10-15 IU/mL were used as decision point for seropositivity. antimicrobial agents.
Due to the routine vaccination of young children and the absence
of widespread virus circulation, individual antibody concentrations TRIS buffer with surfactant and protein (bovine, goat,
decline over time and there is an increasing number of true mouse) stabilizers. Preservatives: ProClin 950 and ProClin 300.
seropositive individuals who would be classified as seronegative if a
concentration of 10 IU/mL is applied.7-10 Warnings and Precautions
More recent studies have demonstrated that the presence of •
E1 antibodies as shown by immunoblot, corresponds well with • For In Vitro Diagnostic Use
neutralization assay results,11 and can be used as a measure of
seropositivity irrespective of antibody levels, also in individuals with
antibody levels below 10 IU/mL.

1
Safety Precautions P333+P313 If skin irritation or rash occurs: Get
medical advice / attention.
P362+P364 Take off contaminated clothing and wash
 CAUTION: This product contains human-sourced and/or it before reuse.
potentially infectious components. Refer to the REAGENTS section Disposal
of this package insert. No known test method can offer complete P501 Dispose of contents / container in
assurance that products derived from human sources or inactivated accordance with local regulations.
microorganisms will not transmit infection. Therefore, all human-
sourced materials should be considered potentially infectious. It Safety Data Sheets are available at www.abbottdiagnostics.com or
is recommended that these reagents and human specimens be contact your local representative.
handled in accordance with the OSHA Standard on Bloodborne For a detailed discussion of safety precautions during system
Pathogens. Biosafety Level 2 or other appropriate biosafety practices operation, refer to the Alinity ci-series Operations Manual, Section 8.
should be used for materials that contain or are suspected of Reagent Handling
containing infectious agents.12-15 • Upon receipt, gently invert the unopened reagent kit by rotating
The following warnings and precautions apply to: it over and back for a full 180 degrees, 5 times with green label
stripe facing up and then 5 times with green label stripe facing
down. This ensures that liquid covers all sides of the bottles
within the cartridges. During reagent shipment, microparticles
can settle on the reagent septum.
WARNING Contains methylisothiazolone and sodium –– Place a check in the square on the reagent kit to indicate to
azide. others that the inversions have been completed.
H317 May cause an allergic skin reaction. • After mixing, place reagent cartridges in an upright position for
EUH032 Contact with acids liberates very toxic gas. 1 hour before use to allow bubbles that may have formed to
Prevention dissipate.
P261 Avoid breathing mist / vapors / spray. • If a reagent cartridge is dropped, place in an upright position
P272 Contaminated work clothing should not be for 1 hour before use to allow bubbles that may have formed to
allowed out of the workplace. dissipate.
P280 Wear protective gloves / protective • Reagents are susceptible to the formation of foam and bubbles.
clothing / eye protection. Bubbles may interfere with the detection of the reagent level in
Response the cartridge and cause insufficient reagent aspiration that may
P302+P352 IF ON SKIN: Wash with plenty of water. adversely affect results.
P333+P313 If skin irritation or rash occurs: Get For a detailed discussion of reagent handling precautions during
medical advice / attention. system operation, refer to the Alinity ci-series Operations Manual,
P362+P364 Take off contaminated clothing and wash Section 7.
it before reuse. Reagent Storage
Disposal Storage Maximum Additional Storage
P501 Dispose of contents / container in Temperature Storage Time Instructions
accordance with local regulations. Unopened 2 to 8°C Until Store in upright position.
expiration If cartridge does not
The following warnings and precautions apply to: date remain upright, gently
invert the cartridge 10
times and place in an
upright position for 1 hour
WARNING Contains methylisothiazolones and before use.
polyethylene glycol octylphenyl ether. Onboard System 30 days
H317 May cause an allergic skin reaction. Temperature
H319 Causes serious eye irritation. Opened 2 to 8°C Until Store in upright position.
H412 Harmful to aquatic life with long lasting expiration If cartridge does not
effects. date remain upright during
Prevention storage, discard the
cartridge.
P261 Avoid breathing mist / vapors / spray.
P264 Wash hands thoroughly after handling. Do not reuse original
reagent caps or
P272 Contaminated work clothing should not be
replacement caps due to
allowed out of the workplace.
the risk of contamination
P280 Wear protective gloves / protective and potential to
clothing / eye protection. compromise reagent
P273 Avoid release to the environment. performance.
Response
P305+P351+P338 IF IN EYES: Rinse cautiously with water Reagents may be stored on or off the system. If removed from the
for several minutes. Remove contact system, store reagents with new replacement caps in an upright
lenses, if present and easy to do. position at 2 to 8°C. For reagents stored off the system, it is
Continue rinsing. recommended that they be stored in their original trays or boxes to
P337+P313 If eye irritation persists: Get medical ensure they remain upright.
advice / attention. For information on unloading reagents, refer to the Alinity ci-series
P302+P352 IF ON SKIN: Wash with plenty of water. Operations Manual, Section 5.

2
Indications of Reagent Deterioration To ensure consistency in results, recentrifuge specimens prior to
Deterioration of the reagents may be indicated when a calibration testing if
error occurs or a control value is out of the specified range. • they contain fibrin, red blood cells, or other particulate matter
Associated test results are invalid, and samples must be retested. • they require repeat testing.
Assay recalibration may be necessary. NOTE: If fibrin, red blood cells, or other particulate matter are
For troubleshooting information, refer to the Alinity ci-series observed, mix by low speed vortex or by inverting 10 times prior to
Operations Manual, Section 10. recentrifugation.
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INSTRUMENT PROCEDURE Prepare frozen specimens as follows:
The Alinity i Rubella IgG assay file must be installed on the Alinity i • Frozen specimens must be completely thawed before mixing.
analyzer prior to performing the assay. • Mix thawed specimens thoroughly by low speed vortex or by
For detailed information on assay file installation and viewing and inverting 10 times.
editing assay parameters, refer to the Alinity ci-series Operations • Visually inspect the specimens. If layering or stratification is
Manual, Section 2. observed, mix until specimens are visibly homogeneous.
For information on printing assay parameters, refer to the Alinity ci- • If specimens are not mixed thoroughly, inconsistent results may
series Operations Manual, Section 5. be obtained.
For a detailed description of system procedures, refer to the Alinity • Recentrifuge specimens.
ci-series Operations Manual. Recentrifugation of Specimens
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SPECIMEN COLLECTION AND PREPARATION • Transfer specimens to a centrifuge tube and centrifuge at
100 000 g-minutes.
FOR ANALYSIS
• Examples of acceptable time and force ranges that meet this
Specimen Types criterion are listed in the table below.
The specimen types listed below were verified for use with this assay Centrifugation time using alternate RCF values can be calculated
on the ARCHITECT i System. using the following formula:
Other specimen types and collection tube types have not been 100 000 g-minutes
verified with this assay. Minimum Centrifugation time (minutes) =
RCF
Specimen Types Collection Tubes
Recentrifugation Time
Serum Serum (Minutes) RCF (x g) g-Minutes
Serum separator 10 10 000 100 000
Plasma Potassium EDTA 20 5000 100 000
Lithium heparin (plasma 40 2500 100 000
separator)
Sodium heparin RCF = 1.12 × rmax (rpm/1000)2
Lithium heparin RCF - The relative centrifugal force generated during
Sodium citrate* centrifugation.
rpm - The revolutions per minute of the rotor on which
* Liquid anticoagulants may have a dilution effect resulting in lower the specimens are being spun (usually the digital
concentration values for individual specimens. Sodium citrate readout on the centrifuge will indicate the rpm).
specimens may result in a -16.8% bias when compared to serum Centrifugation The time should be measured from the time the
specimens. Time - rotor reaches the required RCF or rpm to the time it
• Performance has not been established for the use of cadaveric begins decelerating.
specimens or the use of bodily fluids other than human serum rmax - Radius of the rotor in millimeters. NOTE: If custom
or plasma. tube adapters (i.e., adapters not defined by the
• The instrument does not provide the capability to verify specimen centrifuge manufacturer) are used, then the radius
types. It is the responsibility of the operator to verify that the (rmax) should be manually measured in millimeters
correct specimen types are used in the assay. and the RCF calculated.
Specimen Conditions g-minutes - The unit of measure for the product of RCF (× g)
• Do not use: and centrifugation time (minutes).
–– heat-inactivated specimens • Transfer clarified specimen to a sample cup or secondary tube
–– pooled specimens for testing. For centrifuged specimens with a lipid layer, transfer
–– grossly hemolyzed specimens only the clarified specimen and not the lipemic material.
–– specimens with obvious microbial contamination Specimen Storage
• For accurate results, serum and plasma specimens should be Specimen storage conditions were verified on the ARCHITECT i
free of fibrin, red blood cells, and other particulate matter. Serum System.
specimens from patients receiving anticoagulant or thrombolytic Maximum
therapy may contain fibrin due to incomplete clot formation. Specimen Storage
• To prevent cross contamination, use of disposable pipettes or Type Temperature Time Special Instructions
pipette tips is recommended. Serum/ 2 to 8°C 14 days Specimens may be
Preparation for Analysis Plasma stored on or off the
• Follow the tube manufacturer’s processing instructions for clot, red blood cells, or
collection tubes. Gravity separation is not sufficient for specimen separator gel.
preparation. If testing will be delayed more than 14 days, remove serum or plasma
• Specimens should be free of bubbles. Remove bubbles with an from clot, red blood cells, or separator gel and store frozen (-10°C or
applicator stick before analysis. Use a new applicator stick for colder).
each specimen to prevent cross contamination. Specimens stored frozen for 1 month showed acceptable
performance.
Avoid multiple freeze/thaw cycles.
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Specimen Shipping
Package and label specimens in compliance with applicable state,
federal, and international regulations covering the transport of clinical
specimens and infectious substances.
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PROCEDURE
Materials Provided
08P46 Alinity i Rubella IgG Reagent Kit
Materials Required but not Provided
• Alinity i Rubella IgG assay file
• 08P4601 Alinity i Rubella IgG Calibrators
• 08P4610 Alinity i Rubella IgG Controls or other control material
• 09P1540 Alinity i Multi-Assay Manual Diluent
• Alinity Trigger Solution
• Alinity Pre-Trigger Solution
• Alinity i-series Concentrated Wash Buffer
For information on materials required for operation of the instrument,
refer to the Alinity ci-series Operations Manual, Section 1.
For information on materials required for maintenance procedures,
refer to the Alinity ci-series Operations Manual, Section 9.
Assay Procedure
For a detailed description of how to run an assay, refer to the Alinity
ci-series Operations Manual, Section 5.
• If using primary or aliquot tubes, refer to the Alinity ci-series
Operations Manual, Section 4 to ensure sufficient specimen is
present.
• To minimize the effects of evaporation, verify adequate sample
cup volume is present prior to running the test.
• Maximum number of replicates sampled from the same sample
cup: 10
–– Priority:
◦◦ Sample volume for first test: 70 µL
◦◦ Sample volume for each additional test from same sample
cup: 20 µL
–– ≤ 3 hours on the reagent and sample manager:
◦◦ Sample volume for first test: 150 µL
◦◦ Sample volume for each additional test from same sample
cup: 20 µL
–– > 3 hours on the reagent and sample manager:
◦◦ Replace with a fresh aliquot of sample.
• Refer to the Alinity i Rubella IgG calibrator package insert and/
or Alinity i Rubella IgG control package insert for preparation and
usage.
• For general operating procedures, refer to the Alinity ci-series
Operations Manual, Section 5.
• For optimal performance, it is important to perform routine
maintenance as described in the Alinity ci-series Operations
Manual, Section 9. Perform maintenance more frequently when
required by laboratory procedures.
Sample Dilution Procedures
Samples with IgG antibodies to rubella value exceeding 500.0 IU/mL
are flagged with the code “>500.0 IU/mL” and may be diluted
with either the Automated Dilution Protocol or the Manual Dilution
Procedure.
Automated Dilution Protocol
The system performs a 1:10 dilution of the sample and automatically
calculates the concentration by multiplying the result by the dilution
factor.
Manual Dilution Procedure
Suggested dilution: 1:10
Add 20 μL of the sample to 180 μL of Alinity i Rubella IgG Calibrator
A or Alinity i Multi-Assay Manual Diluent.
4
The operator must enter the dilution factor in the Specimen or
Control tab of the Create Order screen. The system will use this
dilution factor to automatically calculate the concentration of the
sample and report the result. The result should be >5 IU/mL before
the dilution factor is applied.
If the operator does not enter the dilution factor, the result must be
manually multiplied by the appropriate dilution factor before reporting
the result. If a diluted sample result is less than 5 IU/mL do not
report the result. Rerun using an appropriate dilution.
For detailed information on ordering dilutions, refer to the Alinity ci-
series Operations Manual, Section 5.
Calibration
For instructions on performing a calibration, refer to the Alinity ci-
series Operations Manual, Section 5.
Each assay control must be tested to evaluate the assay calibration.
Once a calibration is accepted and stored, it may be used for 22
days. During this time, all subsequent samples may be tested without
further calibration unless:
• A reagent kit with a new lot number is used.
• Daily quality control results are outside of quality control limits
used to monitor and control system performance.
This assay may require recalibration after maintenance to critical
parts or subsystems or after service procedures have been
performed.
Quality Control Procedures
The recommended control requirement for the Alinity i Rubella IgG
assay is that a single sample of each control level be tested once
every 24 hours each day of use.
Additional controls may be tested in accordance with local, state,
and/or federal regulations or accreditation requirements and your
laboratory’s quality control policy.
To establish statistically-based control limits, each laboratory should
establish its own concentration target and ranges for new control lots
at each clinically relevant control level. This can be accomplished
by assaying a minimum of 20 replicates over several (3-5) days and
using the reported results to establish the expected average (target)
and variability about this average (range) for the laboratory. Sources
of variation that should be included in this study in order to be
representative of future system performance include:
• Multiple stored calibrations
• Multiple reagent lots
• Multiple calibrator lots
• Multiple processing modules (if applicable)
• Data points collected at different times of the day
Refer to published guidelines for information or general control
recommendation, for example Clinical and Laboratory Standards
Institute (CLSI) Document C24-A3 or other published guidelines, for
general quality control recommendations.16
• If more frequent control monitoring is required, follow the
established quality control procedures for your laboratory.
• If quality control results do not meet the acceptance criteria
defined by your laboratory, sample results may be suspect.
Follow the established quality control procedures for your
laboratory. Recalibration may be necessary. For troubleshooting
information, refer to the Alinity ci-series Operations Manual,
Section 10.
• Review quality control results and acceptance criteria following a
change of reagent or calibrator lot.
Quality Control Guidance
Refer to “Basic QC Practices” by James O Westgard, Ph.D. for
guidance on laboratory quality control practices.17
Verification of Assay Claims
For protocols to verify package insert claims, refer to Verification of
Assay Claims in the Alinity ci-series Operations Manual.
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RESULTS
Standardization
The Alinity i Rubella IgG assay is referenced to the World Health
Organization (WHO) 1st International Standard (RUBI-1-94) for Anti-
Rubella Immunoglobulin.
The antibody levels of Rubella IgG for a given specimen may vary
when determined by different assay methods and should not be used
interchangeably. The absence of a reference measurement system
and a well defined measurand for Rubella IgG contribute to the
quantitative disagreement observed between assay methods.18
Calculation
The Alinity i Rubella IgG assay utilizes a 4 Parameter Logistic
Curve fit data reduction method (4PLC, Y-weighted) to generate a
calibration and results.
Interpretation of Results
IU/mL Interpretation
0.0 to 4.9 Negative
5.0 to 9.9 *Grayzone (Equivocal) ll
SPECIFIC PERFORMANCE CHARACTERISTICS
≥10.0 19 Positive Representative performance data are provided in this section. Results
obtained in individual laboratories may vary.
* Specimens with grayzone results may contain low levels of The Alinity i analyzer and the ARCHITECT i System utilize the same
Rubella-specific antibodies. For confirmation, samples may be tested reagents and sample/reagent ratios.
by other adequate assays, e.g. immunoblot for the presence of E1
Unless otherwise specified, all studies were performed on the Alinity
antibodies.
i analyzer.
For details on configuring the Alinity i analyzer to use the grayzone
interpretation, refer to the Alinity ci-series Operations Manual, Precision
Section 2. The grayzone interpretation is an editable parameter and Within-Laboratory Precision
should be utilized per end user requirements. A study was performed based on guidance from CLSI EP05-A2.
Testing was conducted using 3 lots of the Alinity i Rubella IgG
Flags
Reagent Kit, 3 lots of the Alinity i Rubella IgG Calibrators, and 3
Some results may contain information in the Flags field. For a
lots of the Alinity i Rubella IgG Controls and 1 instrument. Three
description of the flags that may appear in this field, refer to the
controls and 5 human serum panels were assayed in a minimum of 2
Alinity ci-series Operations Manual, Section 5.
replicates at 2 separate times per day on 20 different days.23
Measuring Interval Within-Run
Measuring interval is defined as the range of values in IU/mL which (Repeatability) Within-Laboratory (Total)a
meets the limits of acceptable performance for linearity, imprecision, Mean SD %CV
and bias. Sample n (IU/mL) SD %CV (Rangeb) (Rangeb)
The measuring interval of the Alinity i Rubella IgG assay is 0.5 to Negative 360 0.0 0.04 N/A 0.04 N/A
500.0 IU/mL. Control (0.00-0.06)
Positive 360 26.7 0.88 3.3 1.08 4.0
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LIMITATIONS OF THE PROCEDURE Control 1 (1.00-1.15) (3.8-4.3)
• For diagnostic purposes, results should be used in conjunction Positive 360 298.9 12.96 4.3 16.89 5.6
with other data; e.g., symptoms, results of other tests, clinical Control 2 (14.95-19.13) (5.2-6.1)
impressions, etc. Panel A 360 3.7 0.15 4.2 0.18 5.0
• Heterophilic antibodies in human serum can react with reagent (0.16-0.21) (4.3-5.6)
immunoglobulins, interfering with in vitro immunoassays. Patients Panel B 360 11.1 0.67 6.0 0.78 7.0
routinely exposed to animals or to animal serum products can (0.65-0.85) (5.9-7.6)
be prone to this interference, and anomalous values may be Panel C 360 100.2 4.29 4.3 4.84 4.8
observed. Additional information may be required for diagnosis.20 (4.51-5.04) (4.4-5.2)
• Specimens from patients who have received preparations of Panel D 359 140.9 6.40 4.5 8.53 6.1
mouse monoclonal antibodies for diagnosis or therapy may (6.58-10.13) (4.9-7.1)
contain human anti-mouse antibodies (HAMA). Specimens Panel E 359 439.2 24.43 5.6 28.40 6.5
containing HAMA may produce anomalous values when tested (21.83-35.96) (5.1-7.6)
with assay kits (such as Alinity i Rubella IgG) that employ mouse a
monoclonal antibodies.21, 22 Includes within-run, between-run, and between-day variability.
b Maximum and minimum SD or %CV for each reagent lot and
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EXPECTED VALUES instrument combination.
This study was performed on the ARCHITECT i System.
Lower Limits of Measurement
Representative performance data are provided in this section.
A study was performed based on guidance from CLSI EP17-A2.
Results obtained in individual laboratories may vary.
Testing was conducted using 3 lots of the Alinity i Rubella IgG
It is recommended that each laboratory determine its own reference Reagent Kit on each of 2 instruments over a minimum of 3 days. The
range based upon its particular locale and population characteristics. maximum observed Limit of Blank (LoB), Limit of Detection (LoD),
The incidence of Rubella IgG antibodies varies among populations and Limit of Quantitation (LoQ) values are summarized below.24
depending on vaccination practices. In this study, an asymptomatic
IU/mL
population composed of 1253 specimens (fresh and frozen) from
LoBa 0.1
pregnant women, random individuals, and negative samples were
LoDb 0.2
tested. Of these specimens, 935 (75%) were positive, 74 (6%) were
equivocal, and 244 (19%) were negative by ARCHITECT Rubella IgG LoQc 0.5
assay. The distribution of this population is shown below.
5
a The LoB represents the 95th percentile from n ≥ 60 replicates of ARCHITECT Rubella IgG Assay
zero-analyte samples. Days Since Alinity i Rubella IgG (IU/mL) (IU/mL)
b The LoD represents the lowest concentration at which the analyte Panel ID 1st Bleed Cutoff: 10.0 IU/mL Cutoff: 10.0 IU/mL
can be detected with 95% probability based on n ≥ 60 replicates of SCP- 0 0.2 (Negative) 0.1 (Negative)
low-analyte level samples. RUB-002 / 5 0.2 (Negative) 0.2 (Negative)
c The LoQ was determined from n ≥ 60 replicates of low-analyte RP-014
7 0.1 (Negative) 0.1 (Negative)
level samples and is defined as the LoD or the mean concentration 12 0.2 (Negative) 0.3 (Negative)
of the samples with the lowest concentration at which the precision 14 1.1 (Negative) 1.1 (Negative)
is met. 19 24.4 (Positive) 22.2 (Positive)
Linearity 21 38.4 (Positive) 35.4 (Positive)
A study was performed based on guidance from CLSI EP06-A.25 26 53.4 (Positive) 54.1 (Positive)
This assay is linear across the measuring interval of 0.5 to 28 48.2 (Positive) 49.1 (Positive)
500.0 IU/mL. 33 55.6 (Positive) 54.3 (Positive)
35 52.7 (Positive) 48.5 (Positive)
Seroconversion Sensitivity
40 55.5 (Positive) 55.1 (Positive)
To determine the seroconversion sensitivity, 5 seroconversion panels
42 57.3 (Positive) 53.6 (Positive)
obtained from commercial vendors were tested on the Alinity i
3325 0 0.3 (Negative) 0.3 (Negative)
analyzer using the Alinity i Rubella IgG assay. The panel results were
evaluated against a comparator assay and data are summarized in 6 12.2 (Positive) 11.6 (Positive)
the following table. 8 22.6 (Positive) 23.4 (Positive)
13 50.0 (Positive) 47.5 (Positive)
ARCHITECT Rubella IgG Assay
Days Since Alinity i Rubella IgG (IU/mL) (IU/mL) 16 47.9 (Positive) 43.4 (Positive)
Panel ID 1st Bleed Cutoff: 10.0 IU/mL Cutoff: 10.0 IU/mL 21 51.4 (Positive) 49.8 (Positive)
SCP- 0 0.3 (Negative) 0.2 (Negative) 23 54.7 (Positive) 48.7 (Positive)
RUB-001 / 2 0.3 (Negative) 0.2 (Negative) 30 64.4 (Positive) 57.3 (Positive)
RP-001 34 58.8 (Positive) 53.8 (Positive)
7 0.2 (Negative) 0.2 (Negative)
9 0.2 (Negative) 0.2 (Negative) 37 57.4 (Positive) 58.9 (Positive)
14 0.4 (Negative) 0.3 (Negative) 41 56.3 (Positive) 54.6 (Positive)
17 3.6 (Negative) 3.0 (Negative) SCP- 0 0.0 (Negative) 0.0 (Negative)
21 19.2 (Positive) 19.7 (Positive) RUB-004 7 0.0 (Negative) 0.0 (Negative)
24 28.0 (Positive) 27.4 (Positive) 18 5.3 (Grayzone) 4.9 (Negative)
28 35.1 (Positive) 31.1 (Positive) 21 22.0 (Positive) 17.5 (Positive)
31 66.8 (Positive) 59.9 (Positive) 25 41.4 (Positive) 41.7 (Positive)
35 59.3 (Positive) 54.8 (Positive) 28 51.6 (Positive) 46.0 (Positive)
38 45.9 (Positive) 46.9 (Positive) 33 65.1 (Positive) 63.3 (Positive)
42 42.3 (Positive) 41.1 (Positive) 36 58.4 (Positive) 54.2 (Positive)
45 40.9 (Positive) 39.0 (Positive) 39 59.9 (Positive) 62.7 (Positive)
50 40.6 (Positive) 40.8 (Positive) 42 66.3 (Positive) 68.6 (Positive)
SCP- 0 0.1 (Negative) 0.1 (Negative) 46 70.3 (Positive) 71.6 (Positive)
RUB-003 / 3 0.0 (Negative) 0.0 (Negative) 49 65.5 (Positive) 65.1 (Positive)
RP-011 53 66.9 (Positive) 70.2 (Positive)
9 0.1 (Negative) 0.1 (Negative)
12 0.1 (Negative) 0.1 (Negative) 56 66.8 (Positive) 75.4 (Positive)
16 3.0 (Negative) 2.7 (Negative) 60 68.1 (Positive) 69.1 (Positive)
19 27.1 (Positive) 27.3 (Positive)
24 66.3 (Positive) 64.7 (Positive) Interference
27 77.0 (Positive) 79.2 (Positive) This study was performed on the ARCHITECT i System.
31 66.6 (Positive) 64.2 (Positive) A study was performed based on guidance from CLSI EP07-A2.
36 76.8 (Positive) 67.8 (Positive) Potential interference was assessed with the ARCHITECT Rubella
39 73.1 (Positive) 60.2 (Positive) IgG assay and the mean interference was determined to be ≤ 10%
for bilirubin, hemoglobin, total protein, and triglycerides at the levels
43 70.4 (Positive) 73.7 (Positive)
indicated below.26
46 74.8 (Positive) 67.0 (Positive)
50 79.8 (Positive) 70.1 (Positive) Potentially Interfering Substance Concentration
53 77.1 (Positive) 73.0 (Positive) Bilirubin ≤ 20 mg/dL
57 73.5 (Positive) 78.9 (Positive) Hemoglobin ≤ 500 mg/dL
60 67.7 (Positive) 68.9 (Positive) Total protein ≤ 12 g/dL
64 71.6 (Positive) 75.3 (Positive) Triglycerides ≤ 3000 mg/dL
67 68.5 (Positive) 68.4 (Positive)
71 81.7 (Positive) 76.8 (Positive)

6
Other Potential Interferents Initial Relative Sensitivity
Additional studies were performed to evaluate other potential This study was performed on the ARCHITECT i System.
interfering disease states on the ARCHITECT Rubella IgG assay. The initial relative sensitivity was 98.4% (932/947) (95% confidence
Potential Interfering Disease interval: 97.4% to 99.1%).
States % Agreementa Number of specimensb
ANA 100.0 9
Initial Relative Specificity
Epstein-Barr Virus 96.0 25
This study was performed on the ARCHITECT i System.
HAMA 100.0 9 The initial relative specificity was 99.0% (191/193) (95% confidence
Herpes Simplex Virus 100.0 20
interval: 96.3% to 99.9%).
Hyper IgG 100.0 9 Comparison of ARCHITECT Rubella IgG with another commercially-
available Rubella IgG assay.
Hyper IgM 100.0 11
Influenza Vaccinees 100.0 10 Initial Relative Agreement (%)
Measles Virus 100.0 10 Site I Site II Site III Total
Parvovirus B19 100.0 7 Mean 99.5 95.6 96.2 98.5
Rheumatoid Factor 100.0 7 95% CI (98.8-99.9) (91.1-98.2) (92.1-98.6) (97.6-99.1)
Systemic Lupus 100.0 6 N 819/823 151/158 153/159 1123/1140
Erythematosis Initial Relative Sensitivity (%)
Varicella Zoster Virus 100.0 10 Mean 99.4 96.6 95.9 98.4
a
95% CI (98.4-99.8) (92.3-98.9) (91.5-98.5) (97.4-99.1)
Compared to a comparator Rubella IgG assay.
b
N 648/652 143/148 141/147 932/947
Specimens giving equivocal results removed.
Initial Relative Specificity (%)
CDC Panels Mean 100.0 80.0 100.0 99.0
This study was performed on the ARCHITECT i System. 95% CI (98.3-100) (44.4-97.5) (77.9-100) (96.3-99.9)
The following information is from a panel of coded serum N 171/171 8/10a 12/12 191/193
specimens (n=100) provided by the Center for Disease Control
(CDC) and tested in a blind study. The sera panel was titered by NOTE: Specimens giving equivocal results using ARCHITECT and
Hemagglutination Inhibition. The ARCHITECT Rubella IgG assay another commercially-available assay were not included in the
results on this sera panel consists of 82 positive tests on 82 positive calculation of relative agreement, relative sensitivity and relative
sera and 18 negative tests on 18 negative sera. This does not imply specificity.
a Specificity 100% following consensus testing.
an endorsement of the assay by the CDC.
Initial Relative Agreement Consensus Testing
This study was performed on the ARCHITECT i System. This study was performed on the ARCHITECT i System.
The presence of IgG antibodies to rubella virus in 1253 specimens Further evaluation of the 17 discordant samples was performed using
(fresh and frozen) was determined at three sites using the another commercially-available Rubella IgG assay (Comparison
ARCHITECT Rubella IgG assay. In addition, each specimen was Assay). From this testing, two samples were equivocal, 13 were
tested using another commercially available Rubella IgG assay. negative and two were positive.
Specimens with results of ≥ 10.0 IU/mL were considered positive, Comparison Assay
specimens with results of 5.0 - 9.9 IU/mL were considered grayzone + -
and specimens with results of < 5.0 IU/mL were considered negative
to calculate sensitivity and specificity. + 2 0
ARCHITECT Result
Of the 1253 specimens evaluated, 113 were equivocal by - 0 13
ARCHITECT and/or the commercially available assay and removed After this additional testing, the consensus relative sensitivity of the
from analysis. Seventeen specimens yielded discordant results ARCHITECT Rubella IgG assay was 100% (95%CI: 99.4%-100%), the
between ARCHITECT and the commercially available assay. The consensus relative specificity was 100% (95%CI: 98.5%-100%), and
relative agreement was 98.5% (1123/1140) (95% confidence interval: the consensus relative agreement was 100% (95%CI: 99.5%-100%).
97.6% to 99.1%).
Method Comparison
Percent Agreement A study was performed based on guidance from CLSI EP09-A3 using
A study was performed based on guidance from CLSI EP12-A2. the Passing-Bablok regression method.28
A study was performed to determine the negative and positive Correlation Concentration
percent agreement between the Alinity i Rubella IgG and the Units n Coefficient Intercept Slope Range
ARCHITECT Rubella IgG assay by testing a minimum of 186 positive Alinity i Serum IU/mL 109 0.99 -1.85 1.1 2.2-452.8
and 299 negative specimens. The specimens were tested in Rubella
singlicate on 6 Alinity i analyzers using 4 lots of Alinity i Rubella IgG IgG vs
Reagent Kit, 1 lot of Alinity i Rubella IgG Calibrator, and 1 lot Alinity ARCHITECT
Rubella IgG
i Rubella IgG Controls. The specimens were also tested in singlicate
using the ARCHITECT Rubella IgG assay. Eighty-six specimens
on the first reagent lot combination, 81 specimens on the second
reagent lot combination and 92 specimens on the third reagent lot
combination that either showed a grayzone result interpretation on
the Alinity i Rubella IgG assay or the ARCHITECT Rubella IgG assay
or on both assays were not included in the evaluation. The negative
and positive percent agreement was 100%.27

7
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