en

25-OH Vitamin D
system
3L52
49-2152/R1

25-OH Vitamin D
Customer Service: Contact your local representative or find country specific contact information
on www.abbottdiagnostics.com

Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any
deviations from the instructions in this package insert.

Key to symbols used

List Number Control Number
In Vitro Diagnostic Medical Reaction Vessels
Device
Sample Cups
Lot Number
Septum
Expiration Date
Replacement Caps
Serial Number Warning: May cause
an allergic reaction
Store at 2-8°C Contains sodium azide.
Contact with acids
liberates very toxic gas.
Consult instructions for use Contains: Methanol

Assay CD-ROM - WW (excluding US) - Addition E

Manufacturer
Assay CD-ROM - WW (excluding US) - Addition E

See REAGENTS section for a full explanation of symbols used in reagent

component naming.

1
NAME • 1 Bottle (5.9 mL per 100-test bottle/26.3 mL per 500‑test
ARCHITECT 25-OH Vitamin D bottle) biotinylated vitamin  D anti-Biotin IgG (mouse, monoclonal)
acridinium-labeled conjugate complex in BIS-TRIS  HCl  buffer with
INTENDED USE
protein stabilizers (bovine gamma globulin) and detergent. Minimum
The ARCHITECT 25-OH Vitamin D assay is a chemiluminescent
concentration: 1.2 μg/mL anti-Biotin IgG and 0.1 μg/mL vitamin‑D‑biotin.
microparticle immunoassay (CMIA) for the quantitative determination of
Preservative: sodium azide.
25-hydroxyvitamin D (25-OH vitamin D) in human serum and plasma.
The ARCHITECT 25-OH Vitamin D assay is to be used as an aid in the • 1 Bottle (4.9 mL per 100-test bottle/21.2 mL per
assessment of vitamin D sufficiency. 500-test bottle) Assay Diluent containing acetic acid buffer with EDTA.
Preservatives: ProClin 300, ProClin 950.
SUMMARY AND EXPLANATION OF TEST • 1 Bottle (10.0 mL per 100-test bottle/50.9 mL per
Vitamin  D is a fat-soluble steroid prohormone mainly produced 500-test bottle) Pre-Treatment 1 containing triethanolamine methanol
photochemically in the skin from 7-dehydrocholesterol. buffer and 8-anilino-1-naphtalensulfonic acid (ANSA).
Two forms of vitamin D are biologically relevant – vitamin D3 (Cholecalciferol) • 1 Bottle (5.9 mL per 100-test bottle/26.3 mL per
and vitamin D2 (Ergocalciferol). Both vitamins D3 and D2 can be absorbed 500-test bottle) Pre-Treatment 2 containing triethanolamine methanol
from food, with vitamin D2 being an artificial source, but only an estimated buffer and 8-anilino-1-naphtalensulfonic acid (ANSA).
10-20% of vitamin D is supplied through nutritional intake1. Vitamins D3
and D2 can be found in vitamin supplements. Vitamin D is converted Other Reagents
to the active hormone 1,25-(OH)2-vitamin  D (Calcitriol) through two ARCHITECT i Pre-Trigger Solution
hydroxylation reactions. The first hydroxylation converts vitamin D into • Pre-trigger solution containing 1.32% (w/v)
25‑OH vitamin D and occurs in the liver. The second hydroxylation converts hydrogen peroxide.
25-OH vitamin D into the biologically active 1,25-(OH)2-vitamin D and ARCHITECT i Trigger Solution
occurs in the kidneys as well as in many other cells of the body. Most • Trigger solution containing 0.35N sodium
cells express the vitamin D receptor and about 3% of the human genome hydroxide.
is directly or indirectly regulated by the vitamin D endocrine system1. ARCHITECT i Wash Buffer
The major storage form of vitamin D is 25-OH vitamin D and is present • Wash buffer containing phosphate buffered saline
in the blood at up to 1,000 fold higher concentration compared to the solution. Preservatives: antimicrobial agents.
active 1,25-(OH)2-vitamin D. 25-OH vitamin D has a half-life of 2-3 weeks
vs. 4 hours for 1,25-(OH)2-vitamin D. Therefore, 25-OH vitamin D is the WARNINGS AND PRECAUTIONS
analyte of choice for determination of the vitamin D status2,3. •
Epidemiological studies have shown a high global prevalence of vitamin D • For In Vitro Diagnostic Use.
insufficiency and deficiency4. The measurement of vitamin D status Package insert instructions must be carefully followed. Reliability of
provides opportunities for preventive and therapeutic interventions5,6,7. assay results cannot be guaranteed if there are any deviations from the
Vitamin D deficiency is a cause of secondary hyperparathyroidism and instructions in this package insert.
diseases related to impaired bone metabolism (like rickets, osteoporosis, Safety Precautions
osteomalacia)2,8,9. Reduced 25-OH vitamin D concentrations in blood
• CAUTION: This product requires the handling of human specimens.
(vitamin D insufficiency) have been associated with an increasing risk
It is recommended that all human-sourced materials be considered
of many chronic illnesses, including common cancers, autoimmune or
potentially infectious and handled in accordance with the OSHA
infectious diseases or cardiovascular problems1,2,6,8,10-12.
Standard on Bloodborne Pathogens.13 Biosafety Level 214 or other
BIOLOGICAL PRINCIPLES OF THE PROCEDURE appropriate biosafety practices15,16 should be used for materials that
The ARCHITECT 25-OH Vitamin D assay is a delayed one-step contain or are suspected of containing infectious agents.
immunoassay including a sample pre-treatment for the quantitative • This product contains sodium azide; for a specific listing, refer to the
determination of vitamin D in human serum and plasma using CMIA REAGENTS section. Contact with acids liberates very toxic gas. This
technology with flexible assay protocols, referred to as Chemiflex. material and its container must be disposed of in a safe way.
Sample and pre-treatment reagent are combined. An aliquot of the • The following warnings and precautions apply to these components:
pre‑treated sample is combined with assay diluent and paramagnetic • Microparticles
anti‑vitamin D coated microparticles to create a reaction mixture. Vitamin D • Assay Diluent
present in the sample binds to anti-vitamin D coated microparticles. After
incubation a biotinylated vitamin D anti-Biotin acridinium-labeled conjugate WARNING: Contains methylisothiazolones.
complex is added to the reaction mixture and binds to unoccupied binding H317 May cause an allergic skin reaction.
sites of the anti-vitamin D coated microparticles. After washing, pre-trigger
and trigger solutions are added to the reaction mixture. The resulting Prevention
chemiluminescent reaction is measured as relative light units (RLUs). An P261 Avoid breathing mist/vapours/spray.
indirect relationship exists between the amount of vitamin D in the sample P272 Contaminated work clothing should not be
and the RLUs detected by the ARCHITECT i System optics. allowed out of the workplace.
For additional information on system and assay technology, refer to the P280 Wear protective gloves/protective clothing/eye
ARCHITECT System Operations Manual, Section 3. protection.
REAGENTS Response
Reagent Kit, 100 Tests/500 Tests
P302+P352 IF ON SKIN: Wash with plenty of water.
NOTE: Some kit sizes are not available in all countries or for use on all
ARCHITECT i Systems. Please contact your local distributor. P333+P313 If skin irritation or rash occurs: Get medical
advice/attention.
ARCHITECT Reagent Kit (3L52-25 or 3L52-35)
P363 Wash contaminated clothing before reuse.
• 1 Bottle (6.6 mL per 100-test bottle/27.0 mL per
500-test bottle) Anti-human vitamin D IgG (sheep, polyclonal) coated This material and its container must be disposed of in a safe way.
microparticles in TRIS buffer. Minimum concentration: 0.05% solids.
Preservatives: ProClin 300, ProClin 950.

2
• The following warnings and precautions apply to these components: Storage Instructions
• Pre-Treatment 1
• Pre-Treatment 2 • The ARCHITECT 25-OH Vitamin D Reagent Kit must be
DANGER: Contains methanol. stored at 2-8°C in an upright position and may be used immediately
after removal from 2-8°C storage.
H226 Flammable liquid and vapour.
• When stored and handled as directed, reagents are stable until the
H332 Harmful if inhaled. expiration date.
H370 Causes damage to organs. • The conjugate is temperature sensitive, minimize room temperature
Prevention exposure of conjugate bottle.
P210 Keep away from heat/sparks/open flames/hot • The ARCHITECT 25-OH Vitamin D 100-test Reagent Kit may be stored
surfaces. No smoking. on board the ARCHITECT i System for a maximum of 7 days.
P243 Take precautionary measures against static • The ARCHITECT 25-OH Vitamin D 500-test Reagent Kit may be stored
discharge. on board the ARCHITECT i System for a maximum of 14 days.
P233 Keep container tightly closed. • After these maximum storage times, the reagent kit must be discarded.
For information on tracking onboard time, refer to the ARCHITECT
P271 Use only in a well-ventilated area. System Operations Manual, Section 5.
P260 Do not breathe mist/vapours/spray. • Reagents may be stored on or off the ARCHITECT i System. If reagents
P280 Wear protective gloves/protective clothing/eye are removed from the system, store them at 2-8°C (with septums and
protection. replacement caps) in an upright position. For reagents stored off the
P264 Wash hands thoroughly after handling. system, it is recommended that they be stored in their original trays and
Response boxes to ensure they remain upright. If the microparticle bottle does
not remain upright (with a septum installed) while in refrigerated
P304+P340 IF INHALED: Remove victim to fresh air and storage off the system, the reagent kit must be discarded. For
keep at rest in a position comfortable for information on unloading reagents, refer to the ARCHITECT System
breathing. Operations Manual, Section 5.
P303+P361+ IF ON SKIN (or hair): Remove/Take off
P353 immediately all contaminated clothing. Rinse Indications of Reagent Deterioration
skin with water/shower. When a control value is out of the specified range, it may indicate
deterioration of the reagents or errors in technique. Associated test
P307+P311 IF EXPOSED: Call a POISON CENTER or
results are invalid and samples must be retested. Assay recalibration may
doctor/physician.
be necessary. For troubleshooting information, refer to the ARCHITECT
Storage System Operations Manual, Section 10.
P403+P235 Store in a well-ventilated place. Keep cool.
INSTRUMENT PROCEDURE
P405 Store locked up.
• The ARCHITECT 25-OH Vitamin D assay requires:
This material and its container must be disposed of in a safe way. • ARCHITECT i System e-Assay CD-ROM found on
• For information on the safe disposal of sodium azide and a detailed www.abbottdiagnostics.com under Support/Technical Library/
discussion of safety precautions during system operation, refer to the Assay Files or
ARCHITECT System Operations Manual, Section 8. • ARCHITECT i System Assay CD ROM - WW (excluding US) -
Handling Precautions Addition E.
• Do not use reagent kits beyond the expiration date. • The ARCHITECT 25-OH Vitamin D assay file must be installed on
• Do not pool reagents within a kit or between reagent kits. the ARCHITECT i System before performing the assay. For detailed
information on assay file installation and on viewing and editing assay
• The conjugate is temperature sensitive, minimize room temperature
parameters, refer to the ARCHITECT System Operations Manual,
exposure of conjugate bottle.
Section 2.
• Before loading the ARCHITECT 25-OH Vitamin D Reagent Kit on
• For information on printing assay parameters, refer to the ARCHITECT
the system for the first time, the microparticle bottle requires mixing
System Operations Manual, Section 5.
to resuspend microparticles that have settled during shipment. For
microparticle mixing instructions, refer to the PROCEDURE, Assay • For a detailed description of system procedures, refer to the
Procedure section of this package insert. ARCHITECT System Operations Manual.
• Septums MUST be used to prevent reagent evaporation and • The default result unit for the ARCHITECT 25-OH Vitamin D assay is
contamination and to ensure reagent integrity. Reliability of assay ng/mL. An alternate result unit, nmol/L, may be selected for reporting
results cannot be guaranteed if septums are not used according to results by editing assay parameter “Result concentration units” to
the instructions in this package insert. nmol/L. The conversion factor used by the system is 2.5.
• To avoid contamination, wear clean gloves when placing a septum on SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS
an uncapped reagent bottle. Specimen Types
• Once a septum has been placed on the reagent bottle, do not The specimen collection tubes listed below were verified to be used with
invert the bottle as this will result in reagent leakage and may the ARCHITECT 25-OH Vitamin D assay. Other specimen collection tubes
compromise assay results. have not been tested with this assay.
• Over time, residual liquids may dry on the septum surface. These • Human serum (including serum collected in serum separator tubes)
are typically dried salts, which have no effect on assay efficacy. • Human plasma collected in:
• For a detailed discussion of handling precautions during system • Potassium-EDTA
operation, refer to the ARCHITECT System Operations Manual,
Section 7. • Lithium heparin (powder or gel)
• Sodium heparin
• Human plasma collected in sodium citrate tube types showed a mean
bias of -13% versus serum.
• Other liquid anticoagulants may also have a dilution effect resulting in
lower concentrations for individual patient specimens.
• The ARCHITECT i System does not provide the capability to verify
specimen type. It is the responsibility of the operator to verify that the
correct specimen types are used in the ARCHITECT 25-OH Vitamin D
assay.

3
Specimen Conditions
• Do not use specimens with the following conditions:
• heat-inactivated
• pooled
• grossly hemolyzed (> 500 mg/dL hemoglobin)
• obvious microbial contamination
• cadaver specimens or any other body fluids
• For accurate results, serum and plasma specimens should be free of
fibrin, red blood cells, and other particulate matter. Serum specimens
from patients receiving anticoagulant or thrombolytic therapy may
contain fibrin due to incomplete clot formation.
• Use caution when handling patient specimens to prevent cross
contamination. Use of disposable pipettes or pipette tips is
recommended.
• For optimal results, inspect all specimens for bubbles. Remove bubbles
with an applicator stick before analysis. Use a new applicator stick for
each specimen to prevent cross contamination.
Preparation for Analysis
• Follow the tube manufacturer’s processing instructions for serum
and plasma collection tubes. Gravity separation is not sufficient for
specimen preparation.
• Mix thawed specimens thoroughly by low speed vortexing or by
inverting 10 times. Visually inspect the specimens. If layering or
stratification is observed, continue mixing until specimens are visibly
homogeneous.
• To ensure consistency in results, specimens must be transferred to a
centrifuge tube and centrifuged at ≥ 10000 RCF (Relative Centrifugal
Force) for 10 minutes before testing if
• they contain fibrin, red blood cells, or other particulate matter,
• they require repeat testing, or
• they were frozen and thawed.
Transfer clarified specimen to a sample cup or secondary tube for
testing.
• Centrifuged specimens with a lipid layer on the top must be transferred
to a sample cup or secondary tube. Care must be taken to transfer
only the clarified specimen without the lipemic material.
Storage
• Specimens may be stored on or off the clot or red blood cells for:
• up to 12 days at 2-8°C or
• up to 72 hours at 15-30°C
• If testing will be delayed more than 12 days, remove serum or plasma
from the clot or red blood cells and store frozen at -20°C or colder.
• Specimens stored frozen for 1 month showed no performance
difference. Avoid more than 3 freeze/thaw cycles.
Shipping
• Before shipping specimens, it is recommended that specimens be
removed from the clot, red blood cells, or separator gel.
• When shipping specimens, package and label specimens in compliance
with applicable state, federal, and international regulations covering
the transport of clinical specimens and infectious substances.
• Specimens may be shipped: ambient, on wet ice, on dry ice. Do not
exceed the storage time limitations listed above.
PROCEDURE
Materials Provided
• 3L52-25 or -35 ARCHITECT 25-OH Vitamin D Reagent Kit
Materials Required but not Provided
• ARCHITECT i System
• 1L66 ARCHITECT i System
Assay CD-ROM - WW (excluding US) - Addition E  , Version 8.0 or
higher or
• ARCHITECT i System e-Assay CD-ROM found on
www.abbottdiagnostics.com under Support/Technical Library/Assay
Files.
4
• 3L52-01 ARCHITECT 25-OH Vitamin D Calibrators
• 3L52-10 ARCHITECT 25-OH Vitamin D Controls
• ARCHITECT i
• ARCHITECT i
• ARCHITECT i
• ARCHITECT i
• ARCHITECT i
• ARCHITECT i
• ARCHITECT i
• Pipettes or pipette tips (optional) to deliver the volumes specified on
the patient or control order screen.
For information on materials required for maintenance procedures, refer to
the ARCHITECT System Operations Manual, Section 9.
Assay Procedure
• Before loading the ARCHITECT 25-OH Vitamin D Reagent Kit on the
system for the first time, the microparticle bottle requires mixing to
resuspend microparticles that have settled during shipment. After the
first time the microparticles have been loaded, no further mixing is
required.
• Invert the microparticle bottle 30 times.
• Visually inspect the bottle to ensure microparticles are
resuspended. If microparticles remain adhered to the bottle,
continue inverting the bottle until the microparticles have been
completely resuspended.
• If the microparticles do not resuspend, DO NOT USE. Contact
your local Abbott representative.
• Once the microparticles have been resuspended, place a septum
on the bottle. For instructions on placing septums on bottles refer
to the Handling Precautions section of this package insert.
• Load the ARCHITECT 25-OH Vitamin D Reagent Kit on the ARCHITECT
i System.
• Verify that all necessary assay reagents are present.
• Ensure that septums are present on all reagent bottles.
• Order calibration, if necessary.
• For information on ordering calibrations, refer to the ARCHITECT
System Operations Manual, Section 6.
• Order tests.
• For information on ordering patient specimens and controls and
for general operating procedures, refer to the ARCHITECT System
Operations Manual, Section 5.
• The minimum sample cup volume is calculated by the system and
is printed on the Orderlist report. No more than 10 replicates may
be sampled from the same sample cup. To minimize the effects of
evaporation, verify adequate sample cup volume is present before
running the test.
• Priority: 60 μL for the first ARCHITECT 25-OH Vitamin D test plus
10 μL for each additional ARCHITECT 25-OH Vitamin D test from
the same sample cup.
• ≤ 3 hours on board: 150 µl for the first ARCHITECT 25-OH
Vitamin D test plus 10 µl for each additional ARCHITECT 25-OH
Vitamin D test from the same sample cup.
• > 3 hours on board: additional sample volume is required. For
information on sample evaporation and volumes, refer to the
ARCHITECT System Operations Manual, Section 5.
• If using primary or aliquot tubes, use the sample gauge to ensure
sufficient patient specimen is present.
• Prepare calibrators and controls.
• Mix ARCHITECT 25-OH Vitamin D Calibrators and Controls by
gentle inversion before use.
• To obtain the recommended volume requirements for the
ARCHITECT 25-OH Vitamin D Calibrators and Controls, hold the
bottles vertically and dispense 5 drops of each calibrator or
5 drops of each control into each respective sample cup.
• Load samples.
• For information on loading samples, refer to the ARCHITECT
System Operations Manual, Section 5.
• Press RUN. • Heterophilic antibodies in human serum can react with reagent
• For additional information on principles of operation, refer to the immunoglobulins, interfering with in vitro immunoassays.17 Patients
ARCHITECT Operations Manual, Section 3. routinely exposed to animals or to animal serum products can be
• For optimal performance, it is important to perform routine maintenance prone to this interference and anomalous values may be observed.
as described in the ARCHITECT System Operations Manual, Section 9. Additional information may be required for diagnosis.
Perform maintenance more frequently when required by laboratory • Specimens from patients who have received preparations of mouse
procedures. monoclonal antibodies for diagnosis or therapy may contain human
anti-mouse antibodies (HAMA).18,19 Specimens containing HAMA
Specimen Dilution Procedures may produce anomalous values when tested with assay kits that
Specimens with a 25-OH vitamin D concentration > 160.0 ng/mL employ mouse monoclonal antibodies.18
(> 400.0 nmol/L) will be flagged as “>160.0 ng/mL” (“>400.0 nmol/L”) and
may be diluted with a specimen with a low 25-OH vitamin D concentration EXPECTED VALUES
≤ 10.7 ng/mL (≤ 26.8 nmol/L). It is recommended that each laboratory establish its own reference
Manual dilutions should be performed as follows: Add 100 μL of the range, which may be unique to the population it serves depending upon
patient specimen to 100 μL of a specimen with a low 25-OH vitamin D geographical, season, patient, dietary, or environmental factors.
concentration (i.e. 1:2 dilution). A reference range study was conducted based on guidance from the
Concentration calculations are performed as follows: Clinical Laboratory and Standards Institute (CLSI), Protocol C28-A320.
Concentration (high specimen) = Serum specimens of apparently healthy individuals from the Netherlands,
collected during summertime (June to September) and wintertime (March
Concentration (1:2 diluted specimen) x 2 - Concentration (low specimen) to May, November and December) were evaluated in replicates of one
Calibration using the ARCHITECT 25-OH Vitamin D assay. A medical questionnaire
• The ARCHITECT 25-OH Vitamin D assay requires a 7-days has been used to select the reference sample group based on the following
recalibration interval. inclusion criteria: age between 17 and 60 years, normal nutrition habits
• To perform an ARCHITECT 25-OH Vitamin D calibration, test (no diet), no food supplements, no bone disease or rheumatism, no recent
calibrators A, B, C, D, E, and F in replicates of two. A single sample sunny vacation or sunbath, no current hospital treatment, no chronic
of each 25-OH vitamin D control level must be tested to evaluate disease (especially diabetes, renal failure, high cholesterol). In addition,
the assay calibration. Ensure that assay control values are within individuals have been excluded when showing results out of the expected
the concentration ranges specified in the control package insert. values for i PTH (15.0 – 68.3 pg/mL) and calcium (2.10 – 2.55 mmol/L).
Calibrators should be priority loaded. The observed values are summarized in the following table.*
• Calibration Range: 0.0 to 160.0 ng/mL (0.0 to 400.0 nmol/L). Season
• For detailed information on how to perform an assay calibration, refer Gender Summer- Winter-
to the ARCHITECT System Operations Manual, Section 6. All Female Male time time Unit
QUALITY CONTROL PROCEDURES n 360 178 182 120 240
The recommended control requirement for the ARCHITECT 25-OH 2.5th Percentile 9.5 9.4 9.4 15.7 8.8 ng/mL
Vitamin  D assay is that a single sample of each control be tested 97.5th Percentile 55.5 59.1 52.4 60.3 46.3 ng/mL
once every 24 hours each day of use. If your laboratory quality control
procedures require more frequent use of controls to verify test results, * Representative data; results in individual laboratories and in different
follow those procedures. geographical areas may vary from these data.
Additional controls may be tested in conformance with local, state, There is currently debate over the recommended target range of vitamin D
and/or federal regulations or accreditation requirements and your in serum, but an expert panel recently suggested a target range of at least
laboratory´s quality control procedure. 30 - 40 ng/mL21.
Each laboratory should establish control ranges to monitor the acceptable SPECIFIC PERFORMANCE CHARACTERISTICS
performance of the assay. If a control is out of its specified range, Data in the section SPECIFIC PERFORMANCE CHARACTERISTICS were
the associated test results are invalid and samples must be retested. generated using the ARCHITECT i2000SR System.
Recalibration may be indicated. Assay results obtained in individual laboratories may vary from data
Verification of Assay Claims presented.
For protocols to verify package insert claims, refer to the ARCHITECT Precision
System Operations Manual, Appendix B. The ARCHITECT 25-OH Vitamin D The ARCHITECT 25-OH Vitamin D assay is designed to have an assay
assay belongs to method group 2. precision of ≤ 10% total %CV.
RESULTS A study was performed with the ARCHITECT 25-OH Vitamin D assay based
The ARCHITECT 25-OH Vitamin D assay uses a 4 Parameter Logistic on guidance from the CLSI Protocol EP5-A222. Six samples consisting of
Curve fit (4PLC, Y-weighted) data reduction method to generate a 3 ARCHITECT 25-OH Vitamin D Controls and 3 serum based panels were
calibration curve. assayed, using two lots of reagents, on one instrument, in replicates of
two at two separate times per day for 20 days. Data from this study are
Calculation
summarized in the following table.
The ARCHITECT i System calculates the Calibrator A through F mean
chemiluminescent signal from two Calibrator A through F replicates, Mean
generates a calibration curve and stores the result. The default result unit Reagent Conc. Within Run Total
for the ARCHITECT 25-OH Vitamin D assay is ng/mL. Sample Lot n (ng/mL) SD %CV SD %CV
Flags Low 1 80 19.0 0.709 3.7 0.712 3.8
Some results may contain information in the Flags field. For a description Control 2 80 19.5 0.589 3.0 0.889 4.6
of the flags that may appear in this field, refer to the ARCHITECT System Medium 1 80 38.5 0.873 2.3 1.142 3.0
Operations Manual, Section 5. Control 2 80 38.0 0.879 2.3 1.062 2.8
Measuring Interval (Reportable Range) High 1 80 78.4 1.470 1.9 2.201 2.8
The measuring interval of the ARCHITECT 25-OH Vitamin D assay is Control 2 80 76.3 1.485 1.9 2.034 2.7
8.0 ng/mL to 160.0 ng/mL (20.0 nmol/L to 400.0 nmol/L). Serum 1 80 23.0 0.714 3.1 0.912 4.0
LIMITATIONS OF THE PROCEDURE Panel 1 2 80 22.4 0.548 2.4 0.780 3.5
• If the vitamin D results are inconsistent with clinical evidence, Serum 1 80 42.5 1.095 2.6 1.346 3.2
additional testing is suggested to confirm the result. Panel 2 2 80 40.1 0.668 1.7 1.274 3.2
• For diagnostic purposes, results should be used in conjunction with Serum 1 80 75.4 1.088 1.4 2.064 2.7
other data; e.g., symptoms, results of other tests, clinical impressions, Panel 3 2 80 71.3 1.242 1.7 1.869 2.6
etc.

5
Linear Range Potentially Interfering Range of
Based on guidance from the NCCLS Protocol EP6-A23 a study was Substance Concentration % Recoverya
performed to establish the linear range of the ARCHITECT 25-OH Hemoglobinb 200 mg/dL 91 - 98
Vitamin D assay.
Bilirubin 20 mg/dL 100 - 104
Four sample pairs were prepared with 11 dilutions for each pair by
mixing a high 25-OH vitamin D sample (in the range from 78.4 ng/mL to Triglycerides 5000 mg/dL 94 - 103
165.5 ng/ mL) in specific ratios with a low 25-OH vitamin D sample (in the Protein (Human Albumin) 12 g/dL 90 - 102
range from 0.2 ng/mL to 10.7 ng/mL) and tested using the ARCHITECT Rheumatoid Factorc 400 IU/mL 106 - 109
25-OH Vitamin D assay. Using an absolute deviation from linearity of HAMA 1000 ng/mL 96 - 99
≤ 20% a linear range of 9.4 ng/mL to 165.5 ng/mL was established for the
Red blood cells 0.4 %(v/v) 99 - 103
ARCHITECT 25-OH Vitamin D assay.
Measuring Interval a % Recovery = Mean Observed Value (ng/mL) x 100
Measuring interval is defined as the range of values in ng/mL which Mean Control Value (ng/mL)
meets the limits of acceptable performance for both imprecision and bias b For hemoglobin with concentrations between 200 mg/dL and
for an undiluted sample. For the verification studies described in this 500 mg/dL the % Recovery ranged from 90% to 62%.
package insert, the range was 8.0 ng/mL to 160.0 ng/mL (20.0 nmol/L c For rheumatoid factors with concentrations between 400 IU/mL and
to 400.0 nmol/L).
800 IU/mL the % Recovery ranged from 107% to 118%.
Sensitivity
Method Comparison
The ARCHITECT 25-OH Vitamin D assay is designed to have a Limit of
Blank (LoB) of ≤ 4.0 ng/mL, a Limit of Detection (LoD) of ≤ 10.0 ng/mL The ARCHITECT 25-OH Vitamin D assay is designed to have a correlation
and a Limit of Quantitation (LoQ) of ≤ 20 ng/mL. coefficient of ≥ 0.80 for serum samples when evaluated against the
DiaSorin LIAISON 25-OH Vitamin D Total.
Based on guidance from the CLSI Protocol EP17-A24 a study was
performed with 4 zero-level samples (Calibrator A) and 8 samples with A study was performed with the ARCHITECT 25-OH Vitamin D assay,
25-OH vitamin D concentrations ranging from 3.4 ng/mL to 9.5  ng/ mL. where regression analysis was performed using the Passing-Bablok26
These samples were tested in 5 separate runs over a minimum of 5 days method. Data from this study are summarized in the following table and
using two reagent lots and two instruments. graph.*
In the above described study the LoB was 1.9 ng/mL, the LoD was Regression Correlation
3.1 ng/ mL and the LoQ was 8.0 ng/mL. Method n Slope Intercept Coefficient
Specificity Passing-
The specificity of the ARCHITECT 25-OH Vitamin D assay was assessed Babloka 108 1.02 -0.54 0.94
by testing the cross-reactants listed in the table below. a A linear regression method with no special assumptions regarding the
A study was performed with the ARCHITECT 25-OH Vitamin D assay based distribution of the samples and measurement errors.
on guidance from the CLSI Protocol EP7-A225. Aliquots of ARCHITECT * Representative data; variables such as differences in sampling size and
25-OH Vitamin D Calibrator A were supplemented with potential sample population may impact the correlation of the assay, therefore,
cross-reactants at the concentrations listed and tested for 25-OH vitamin D. results in individual laboratories may vary from these data.
Data from this study are summarized in the following table. In this evaluation, serum specimen concentrations ranged from 8.2 ng/ mL
Concentration % Cross- to 70.5 ng/mL with the ARCHITECT 25-OH Vitamin D assay and from
Cross-Reactant ng/mL Reactivitya 5.1  ng/mL to 63.2 ng/mL with the DiaSorin LIAISON 25-OH Vitamin D
Total. The specimens included in the study are sourced from a clinical
25-OH vitamin D3 100 105
research organization.
25-OH vitamin D2 100 52
Vitamin-D2 (Ergocalciferol) 1000 0.1 ARCHITECT 25-OH Vitamin D
vs.
Vitamin-D3 (Cholecalciferol) 1000 0.3
DiaSorin LIAISON 25-OH Vitamin D Total (Passing-Bablok)
24,25-(OH)2-vitamin D3 20 112
1,25-(OH)2-vitamin D3 100 12.6
3-epi 25-OH vitamin D3 100 2.7
Paricalcitol (Zemplar) 24 0.4

Mean Value spiked (ng/mL) -

a % Cross-Reactivity = Mean Value non spiked (ng/mL) x 100
Concentration of Cross-Reactant (ng/mL)
Interference
Potential interference in the ARCHITECT 25-OH Vitamin  D assay from
hemoglobin, bilirubin, triglycerides, protein, rheumatoid factor, HAMA
(human anti-mouse antibodies) and red blood cells is designed to be
≤ 10%.
Interference was demonstrated by a study based on guidance from
the CLSI Protocol EP7-A2. Data from this study are summarized in the
following table.

DiaSorin LIAISON 25-OH Vitamin D Total (ng/mL)

6
The same specimens have been measured with a chromatographic method
(LC-MS/MS) and evaluated against the ARCHITECT 25-OH Vitamin D
assay. The regression analysis was performed using the Passing-Bablok
method. Data from this study are summarized in the following table and
graph.*
Regression Correlation
Method n Slope Intercept Coefficient
Passing-
Babloka 107 1.01 -1.45 0.90
a A linear regression method with no special assumptions regarding the
distribution of the samples and measurement errors.
* Representative data; variables such as differences in sampling size
and sample population may impact the correlation of the assay,
therefore, results in individual laboratories may vary from these data.
Additionally, LC-MS/MS methodology variables, such as extraction
method, ionization method, type of chromatographic separation and
type of internal standard may impact the LC-MS/MS results; therefore
results in individual laboratories may vary from these data27.
In this evaluation, serum specimen concentrations ranged from
8.2 ng/mL to 70.5 ng/mL with the ARCHITECT 25-OH Vitamin D assay and
from 7.5 ng/mL to 64.4 ng/mL with the LC-MS/MS test method.
ARCHITECT 25-OH Vitamin D
vs.
LC-MS/MS 25-OH Vitamin D (Passing-Bablok)
LC-MS/MS 25-OH Vitamin D (ng/mL)
BIBLIOGRAPHY
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7
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The following US Patents are relevant to the ARCHITECT i System or its
components. There are other such patents and patent applications in the
United States and worldwide.
5 468 646 5 543 524 5 545 739
5 565 570 5 669 819 5 783 699

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