2P13-28

en
2nd Generation Testosterone
ARCHITECT 2P13-23 2P13
2nd Generation Testosterone ABBL421 / R03
B2P1W0
Read Highlighted Changes: Revised November 2016.

Package insert instructions must be carefully followed. Reliability of 2. After incubation, testosterone acridinium-labeled conjugate is
assay results cannot be guaranteed if there are any deviations from added to the reaction mixture.
the instructions in this package insert. 3. After further incubation and washing, Pre-Trigger and Trigger
ll
NAME Solutions are added to the reaction mixture.
4. The resulting chemiluminescent reaction is measured as relative
ARCHITECT 2nd Generation Testosterone
light units (RLUs). There is an inverse relationship between the
ll
INTENDED USE amount of testosterone in the sample and the RLUs detected by
The ARCHITECT 2nd Generation Testosterone assay is a the ARCHITECT iSystem optics.
chemiluminescent microparticle immunoassay (CMIA) for the The concentration of testosterone is interpolated from a calibration
quantitative determination of testosterone in human serum and curve established with calibrators of known testosterone
plasma. Measurements of testosterone are used in the diagnosis and concentration.
treatment of disorders involving the male sex hormones (androgens), For additional information on system and assay technology, refer to
including primary and secondary hypogonadism, delayed or the ARCHITECT System Operations Manual, Section 3.
precocious puberty, impotence in males and, in females, hirsutism
(excessive hair) and virilization (masculinization) due to tumors, ll
REAGENTS
polycystic ovaries, and adrenogenital syndromes. Kit Contents
ll
SUMMARY AND EXPLANATION OF THE TEST ARCHITECT 2nd Generation Testosterone 2P13
Testosterone is regarded as the most important of the androgen NOTE: Some kit sizes are not available in all countries or for use on
steroids. In males, it is secreted by the Leydig and interstitial cells of all ARCHITECT iSystems. Please contact your local distributor.
the testes which are stimulated by luteinizing hormone (LH). Control 2P13-28 2P13-23
of testosterone secretion is through a negative feedback loop to the
hypothalamus where secretion of gonadotrophin-releasing hormone 100 400
promotes synthesis and release of LH and follicle-stimulating
1 x 6.6 mL 4 x 6.6 mL
hormone (FSH) from the anterior pituitary gland.
In females, testosterone is secreted by the follicular theca and 1 x 6.9 mL 4 x 6.9 mL
interstitial cells of the ovaries and also produced by metabolism of 1 x 25.0 mL 4 x 25.0 mL
adrenal androgens. The concentrations of testosterone are typically
1 x 12.2 mL 4 x 12.2 mL
about 10-20 times lower for females than for males.
In the circulation, approximately 97% of testosterone is transported Anti-Testosterone (sheep, monoclonal) coated
by proteins, most notably by binding to sex hormone-binding globulin microparticles in BIS Tris buffer with protein (bovine) stabilizer.
(SHBG) with an affinity of approximately 109 Lmol-1.1 Testosterone is Minimum concentration: 0.1% solids. Preservative: ProClin 300.
also weakly bound to albumin.
Testosterone acridinium-labeled conjugate in BIS Tris
The ARCHITECT 2nd Generation Testosterone assay releases
buffer with surfactant stabilizer. Minimum concentration: 6.5 nmol/L.
testosterone from binding proteins and measures total testosterone.
Preservative: ProClin 300.
Free testosterone can be calculated from the total testosterone,
SHBG and albumin concentrations.2 The Free Androgen Index (FAI) Testosterone Assay Specific Diluent
may also be calculated (FAI = [Total Testosterone] / [SHBG]) and containing phosphate and glycine in citrate buffer. Preservative:
provides an index of free testosterone status. This ratio correlates ProClin 300.
well with both measured and calculated values of free testosterone Testosterone Specimen Diluent containing PBS
and helps to discriminate subjects with excessive androgen activity buffer. Preservative: ProClin 300.
from normal individuals.3-5
The concentration of testosterone in an individual fluctuates over 24 Other Reagents
hours.6 The pulsatile release of LH in the night typically leads to a ARCHITECT Pre-Trigger Solution containing
peak of testosterone concentration in the morning. Time of day, age, 1.32% (w/v) hydrogen peroxide.
sex, puberty, pre- and post-menopause, and disease, all have an
influence on testosterone concentration and should be considered in ARCHITECT Trigger Solution containing 0.35 N
interpreting individual results. sodium hydroxide.

ll
BIOLOGICAL PRINCIPLES OF THE PROCEDURE ARCHITECT Wash Buffer containing phosphate
buffered saline solution. Preservatives: antimicrobial agents.
The ARCHITECT 2nd Generation Testosterone assay is a delayed
one-step immunoassay for the quantitative determination of Warnings and Precautions
testosterone in human serum and plasma using CMIA technology
•
with flexible assay protocols, referred to as Chemiflex.
• For In Vitro Diagnostic Use
1. Sample, assay specific diluent and anti-testosterone (sheep,
monoclonal) coated paramagnetic microparticles are combined.
The testosterone present in the sample binds to the anti-
testosterone coated microparticles.

1
Safety Precautions Prevention
CAUTION: This product requires the handling of human specimens. P234 Keep only in original container.
It is recommended that all human-sourced materials be considered P261 Avoid breathing mist / vapors / spray.
potentially infectious and handled in accordance with the OSHA P272 Contaminated work clothing should not be
Standard on Bloodborne Pathogens. Biosafety Level 2 or other allowed out of the workplace.
appropriate biosafety practices should be used for materials that
P280 Wear protective gloves / protective
contain or are suspected of containing infectious agents.7-10
clothing / eye protection.
The following warnings and precautions apply to: Response
P302+P352 IF ON SKIN: Wash with plenty of water.
P333+P313 If skin irritation or rash occurs: Get
medical advice / attention.
P362+P364 Take off contaminated clothing and wash
WARNING Contains methylisothiazolones and sodium
it before reuse.
azide.
P390 Absorb spillage to prevent material
H317 May cause an allergic skin reaction.
damage.
EUH032 Contact with acids liberates very toxic gas.
Disposal
Prevention
P501 Dispose of contents / container in
P261 Avoid breathing mist / vapors / spray.
accordance with local regulations.
P272 Contaminated work clothing should not be
allowed out of the workplace. Safety Data Sheets are available at [Link] or
P280 Wear protective gloves / protective contact your local representative.
clothing / eye protection. For a detailed discussion of safety precautions during system
Response operation, refer to the ARCHITECT System Operations Manual,
P302+P352 IF ON SKIN: Wash with plenty of water. Section 8.
P333+P313 If skin irritation or rash occurs: Get Reagent Handling
medical advice / attention. • Do not use reagent kits beyond the expiration date.
P362+P364 Take off contaminated clothing and wash • Do not pool reagents within a kit or between kits.
it before reuse. • Before loading the reagent kit on the system for the first time, the
Disposal microparticle bottle requires mixing to resuspend microparticles
P501 Dispose of contents / container in that may have settled during shipment. For microparticle mixing
accordance with local regulations. instructions, refer to the PROCEDURE, Assay Procedure section
of this package insert.
The following warnings and precautions apply to: and
• Septums MUST be used to prevent reagent evaporation and
contamination and to ensure reagent integrity. Reliability of
assay results cannot be guaranteed if septums are not used
according to the instructions in this package insert.
• To avoid contamination, wear clean gloves when placing a
WARNING Contains methylisothiazolones. septum on an uncapped reagent bottle.
H317 May cause an allergic skin reaction. • Once a septum has been placed on an open reagent bottle,
Prevention do not invert the bottle as this will result in reagent leakage
P261 Avoid breathing mist / vapors / spray. and may compromise assay results.
P272 Contaminated work clothing should not be • Over time, residual liquids may dry on the septum surface.
allowed out of the workplace. These are typically dried salts and have no effect on assay
P280 Wear protective gloves / protective efficacy.
clothing / eye protection. For a detailed discussion of handling precautions during system
Response operation, refer to the ARCHITECT System Operations Manual,
P302+P352 IF ON SKIN: Wash with plenty of water. Section 7.
P333+P313 If skin irritation or rash occurs: Get Reagent Storage
medical advice / attention. When stored and handled as directed, reagents are stable until the
P362+P364 Take off contaminated clothing and wash expiration date.
it before reuse. Storage Maximum Additional Storage
Disposal Temperature Storage Time Instructions
P501 Dispose of contents / container in Unopened/ 2-8°C Until May be used immediately
accordance with local regulations. Opened* expiration after removal from 2-8°C
date storage.
The following warnings and precautions apply to:
Store in upright position.
On board System 30 days Discard after 30 days.
temperature For information on tracking
onboard time, refer to
the ARCHITECT System
WARNING Contains hydrochloric acid and Operations Manual,
methylisothiazolones. Section 5.
H317 May cause an allergic skin reaction.
* Reagents may be stored on or off the ARCHITECT iSystem. If
H290 May be corrosive to metals.
reagents are removed from the system, store them at 2-8°C (with
septums and replacement caps) in an upright position. For reagents
stored off the system, it is recommended that they be stored in

2
their original trays and boxes to ensure they remain upright. If the Preparation for Analysis
microparticle bottle does not remain upright (with a septum • Follow the tube manufacturer’s processing instructions for
installed) while in refrigerated storage off the system, the reagent collection tubes. Gravity separation is not sufficient for specimen
kit must be discarded. For information on unloading reagents, refer preparation.
to the ARCHITECT System Operations Manual, Section 5. • Mix thawed specimens thoroughly by low speed vortexing or by
Indications of Reagent Deterioration inverting 10 times. Visually inspect the specimens. If layering or
When a control value is out of the specified range, it may indicate stratification is observed, continue mixing until specimens are
deterioration of the reagents or errors in technique. Associated test visibly homogeneous.
results are invalid, and samples must be retested. Assay recalibration • To ensure consistency in results, specimens must be transferred
may be necessary. For troubleshooting information, refer to the to a centrifuge tube and centrifuged at ≥1,000 RCF (Relative
ARCHITECT System Operations Manual, Section 10. Centrifugal Force) for 10 minutes before testing if
ll
INSTRUMENT PROCEDURE • they contain fibrin, red blood cells, or other particulate
matter or
The ARCHITECT 2nd Generation Testosterone assay file must be
installed on the ARCHITECT iSystem from an ARCHITECT iSystem • they were frozen and thawed.
Assay CD-ROM prior to performing the assay. • Transfer clarified specimen to a sample cup or secondary tube
For detailed information on assay file installation and viewing for testing. For centrifuged specimens with a lipid layer, transfer
and editing assay parameters, refer to the ARCHITECT System only the clarified specimen and not the lipemic material.
Operations Manual, Section 2. • Inspect all specimens for bubbles. Remove bubbles with an
For information on printing assay parameters, refer to the ARCHITECT applicator stick before analysis. Use a new applicator stick for
System Operations Manual, Section 5. each specimen to prevent cross contamination.
For a detailed description of system procedures, refer to the Specimen Storage
ARCHITECT System Operations Manual. Specimen Type Storage Temperature Maximum Storage Time
Alternate Result Units Serum/Plasma Room temperature ≤ 8 hours
The default result unit for the ARCHITECT 2nd Generation 2-8°C ≤ 7 days
Testosterone assay is nmol/L. Alternate result units, ng/dL or ng/ -20°C or colder --
mL, may be selected for reporting results by editing assay parameter
“Result concentration units”, to ng/dL or ng/mL. The conversion Remove the serum from clot or separator gel as soon as possible
factor used by the system is 28.84 for the conversion to ng/dL, and after complete clot formation, or plasma from red blood cells as soon
0.2884 for the conversion to ng/mL. as possible upon receipt. If testing will not be performed within 8
ll
SPECIMEN COLLECTION AND PREPARATION hours of draw, specimens may be stored either at 2-8°C for up to 7
days or frozen (-20°C or colder), prior to being tested.
FOR ANALYSIS
Avoid more than 1 freeze/thaw cycle.
Specimen Types
Specimen Shipping
Verified specimen types to be used with this assay:
• Package and label specimens in compliance with applicable
Specimen Types Collection Tubes
state, federal, and international regulations covering the transport
Serum Serum of clinical specimens and infectious substances.
Serum separator tubes • Do not exceed the storage limitations listed above.
Plasma Sodium heparin
Lithium heparin ll
PROCEDURE
Dipotassium EDTA Materials Provided
2P13 ARCHITECT 2nd Generation Testosterone Reagent Kit
• Lithium heparin plasma separator tubes (PST and PST-II) cannot
be used with the ARCHITECT 2nd Generation Testosterone Materials Required but not Provided
assay. • ARCHITECT 2nd Generation Testosterone Assay file obtained
• Other specimen collection tube types have not been tested with from the ARCHITECT iSystem e-Assay CD-ROM found on
this assay. [Link].
• The instrument does not provide the capability to verify specimen • 2P13-01 ARCHITECT 2nd Generation Testosterone Calibrators
type. It is the responsibility of the operator to verify that the • 2P13-10 ARCHITECT 2nd Generation Testosterone Controls (or
correct specimen types are used in the assay. other control material)
• Performance has not been established for the use of cadaveric • ARCHITECT Pre-Trigger Solution
specimens or the use of body fluids other than human serum • ARCHITECT Trigger Solution
and plasma. • ARCHITECT Wash Buffer
Specimen Conditions • ARCHITECT Reaction Vessels
• Do not use specimens with the following conditions: • ARCHITECT Sample Cups
• heat-inactivated • ARCHITECT Septum
• pooled • ARCHITECT Replacement Caps
• grossly hemolyzed • Pipettes or pipette tips (optional) to deliver the volumes specified
• obvious microbial contamination on the patient or control order screen.
• For accurate results, serum and plasma specimens should be For information on materials required for maintenance procedures,
free of fibrin, red blood cells, and other particulate matter. Serum refer to the ARCHITECT System Operations Manual, Section 9.
specimens from patients receiving anticoagulant or thrombolytic Assay Procedure
therapy may contain fibrin due to incomplete clot formation. • Before loading the reagent kit on the system for the first time, the
• To prevent cross contamination, use of disposable pipettes or microparticle bottle requires mixing to resuspend microparticles
pipette tips is recommended. that may have settled during shipment. After the first time the
microparticles have been loaded, no further mixing is required.

3
 • Invert the microparticle bottle 30 times. • ≤ 3 hours on board: 200 μL for the first ARCHITECT 2nd
• Visually inspect the bottle to ensure microparticles are Generation Testosterone test plus 150 μL for each additional
resuspended. If microparticles are still adhered to the bottle, ARCHITECT 2nd Generation Testosterone test from the same
continue to invert the bottle until the microparticles have sample cup.
been completely resuspended. • If using primary or aliquot tubes, use the sample gauge to ensure
• If the microparticles do not resuspend, DO NOT USE. sufficient patient specimen is present.
Contact your local Abbott representative. • Prepare calibrators and controls.
• Once the microparticles have been resuspended, place a • The ARCHITECT 2nd Generation Testosterone Calibrators
septum on the bottle. For instructions about placing septums and Controls may be thawed at room temperature for 90
on bottles, refer to the Reagent Handling section of this to 120 minutes or overnight at 2-8°C. After thawing, it is
package insert. suggested to record the thaw date on the carton or the
• Load the reagent kit on the ARCHITECT iSystem. bottles as an aid in tracking to the expiration date.
• Verify that all necessary reagents are present. • Mix the ARCHITECT 2nd Generation Testosterone Calibrators
• Ensure that septums are present on all reagent bottles. and all Controls by gentle inversion (10 times) prior to use.
• Order calibration, if necessary. • To obtain the recommended volume requirements for the
• For information on ordering calibrations, refer to the ARCHITECT 2nd Generation Testosterone Calibrators and
ARCHITECT System Operations Manual, Section 6. Controls, hold the bottles vertically, and dispense 10 drops
• Order tests. of each calibrator or 10 drops of the low control, and 5 drops
of the medium and high control into each respective sample
• For information on ordering patient specimens and
cup.
controls and for general operating procedures, refer to the
ARCHITECT System Operations Manual, Section 5. • Follow the manufacturer’s instructions for preparation of
commercially available control material.
• Select the appropriate assay protocol for control and specimen
testing. NOTE: It is very important to return the ARCHITECT 2nd
Generation Testosterone Calibrators and Controls to the
• Default Assay Dilution Protocol
correct storage conditions immediately after use, as follows:
The 1:3 dilution protocol is the default protocol for all
• Return ARCHITECT 2nd Generation Testosterone Calibrators
patient samples and the medium and high controls. In this and Controls to their carton and store at 2-8°C.
protocol, samples are diluted 1:3 with Specimen Diluent. If
• Load samples.
sample results are greater than 35 nmol/L, the instrument
• For information on loading samples, refer to the ARCHITECT
automatically orders a retest using the 1:4 dilution protocol.
System Operations Manual, Section 5.
If sample results are less than 0.45 nmol/L, the instrument
• Press RUN.
automatically orders a retest using the neat (undiluted)
• For additional information on principles of operation, refer to the
assay protocol. Laboratories may also choose to override the
ARCHITECT System Operations Manual, Section 3.
default for any sample with a result less than 1.0 nmol/L.
• For optimal performance, it is important to perform routine
• 1:4 Assay Dilution Protocol maintenance as described in the ARCHITECT System Operations
The 1:4 dilution protocol is an alternate dilution protocol on Manual, Section 9. Perform maintenance more frequently when
the instrument. In this protocol, samples are diluted 1:4 with required by laboratory procedures.
Specimen Diluent. If sample results generated by the default
Samples Dilution Procedures
protocol (1:3) are greater than 35 nmol/L, the instrument
Samples with testosterone serum or plasma value exceeding
automatically orders a retest using this protocol. Laboratories
35 nmol/L are flagged with the code “ > 35” and may be diluted
may also choose to override the default for any sample with
using the Automated Dilution Protocol.
a result greater than 35 nmol/L.
Automated Dilution Protocol
• Neat (Undiluted) Assay Protocol
The system performs a 1:4 dilution of the sample with the Specimen
The neat protocol tests samples undiluted. This protocol is Diluent and automatically calculates the concentration of the
utilized for patient samples with results between 0.00 and specimen before dilution and reports the result.
1.0 nmol/L, and the low control. Samples may be tested For detailed information on ordering dilutions, refer to the ARCHITECT
using this protocol initially or if the result produced is less System Operations Manual, Section 5.
than 0.45 nmol/L.
Calibration
• The minimum sample cup volume is calculated by the system
• Test Calibrators A-F in duplicate. The calibrators should be
and is printed on the Orderlist report. No more than 10 replicates
priority loaded.
may be sampled from the same sample cup. To minimize the
effects of evaporation, verify adequate sample cup volume is A single replicate of each control level must be tested to
present prior to running the test. evaluate the assay calibration. Ensure that assay control values
are within the ranges specified in the respective control package
• For the default dilution protocol (1:3) / dilution protocol (1:4):
insert.
• Priority: 100 μL / 88 μL for the first ARCHITECT 2nd
• Calibration Range: 0.0 - 30.0 nmol/L.
Generation Testosterone test plus 50 μL / 38 μL for each
additional ARCHITECT 2nd Generation Testosterone test • Once an ARCHITECT 2nd Generation Testosterone calibration
from the same sample cup. is accepted and stored, all subsequent samples may be tested
without further calibration unless:
• ≤ 3 hours on board: 150 μL for the first ARCHITECT 2nd
Generation Testosterone test plus 50 μL for each additional • A reagent kit with a new lot number is used or
ARCHITECT 2nd Generation Testosterone test from the same • Controls are out of range.
sample cup. • For detailed information on how to perform an assay calibration,
• For the neat (undiluted) protocol: refer to the ARCHITECT System Operations Manual, Section 6.
• Priority: 200 μL for the first ARCHITECT 2nd Generation
Testosterone test plus 150 μL for each additional ARCHITECT
2nd Generation Testosterone test from the same sample
cup.

4
Quality Control Procedures Category Age Testosterone nmol/L (ng/dL)
The recommended control requirement for the ARCHITECT 2nd Apparently Range 5th 95th
Generation Testosterone assay is that a single replicate of each Healthy n (years) Median Min. Max. percentile percentile
control level be tested once every 24 hours each day of use. If the Males 129 21-49 17.13 1.63 34.00 8.33 30.19
quality control procedures in your laboratory require more frequent (21-49 years (494.03) (47.01) (980.56) (240.24) (870.68)
use of controls to verify test results, follow your laboratory-specific of age)
procedures. Males 71 50-77 15.34 4.41 35.38 7.66 24.82
Additional controls may be tested in accordance with local, state, (≥50 years of (442.41) (127.18) (1020.36) (220.91) (715.81)
age)
and/or federal regulations or accreditation requirements and your
laboratory’s quality control policy. Females 129 21-49 0.86 0.25 2.75 0.48 1.85
(21-49 years (24.80) (7.21) (79.31) (13.84) (53.35)
Each laboratory should establish control ranges to monitor the of age)
acceptable performance of the assay. If a control is out of its Females 52 50-82 0.82 0.30 1.28 0.43 1.24
specified range, the associated sample results are invalid and the (≥50 years of (23.50) (8.65) (36.92) (12.40) (35.76)
samples must be retested. Recalibration may be indicated. age)
Verification of Assay Claims
Expected ranges for pediatric samples were obtained for the
For protocols to verify package insert claims, refer to the ARCHITECT
ARCHITECT 2nd Generation Testosterone assay from testing a
System Operations Manual, Appendix B.
minimum of 200 samples divided by males 7-18 years of age (n=120)
The ARCHITECT 2nd Generation Testosterone assay belongs to and females 7-18 years of age (n=116). The individuals included
method group 6. in the study had no clinical or endocrinological signs of premature
ll
RESULTS or delayed puberty or virilization. The data are summarized in the
following tables.
Calculation
Males nmol/L (ng/dL)
The ARCHITECT 2nd Generation Testosterone assay utilizes a
4 Parameter Logistic Curve fit data reduction method (4PLC, 5th 95th
Stage n Median Min. Max. percentile percentile
Y-weighted) to generate a calibration curve.
Tanner Stage I 22 0.28 0.08 1.05 0.09 1.02
Flags (7.93) (2.31) (30.28) (2.52) (29.29)
Some results may contain information in the Flags field. For a Tanner Stage II 23 0.87 0.13 9.78 0.13 9.66
description of the flags that may appear in this field, refer to the (25.09) (3.75) (282.06) (3.86) (278.59)
ARCHITECT System Operations Manual, Section 5. Tanner Stage III 25 0.86 0.30 23.64 0.31 22.73
Measuring Interval (Reportable Range) (24.80) (8.65) (681.78) (8.91) (655.39)
Tanner Stage IV 25 7.62 0.62 27.24 0.69 26.16
Measuring interval is defined as the range of values in nmol/L which
(219.76) (17.88) (785.60) (19.96) (754.45)
meets the acceptable performance for both imprecision and bias
Tanner Stage V 25 18.44 0.46 31.42 0.58 31.28
across all available assay file dilutions. The range was 0.15 nmol/L
(531.81) (13.27) (906.15) (16.64) (902.09)
(Limit of Quantitation - LoQ) to 64.57 nmol/L.
ll
LIMITATIONS OF THE PROCEDURE
Females nmol/L (ng/dL)
5th 95th
• Lithium heparin plasma separator tubes (PST and PST-II) cannot Stage n Median Min. Max. percentile percentile
be used with the ARCHITECT 2nd Generation Testosterone Tanner Stage I 25 0.28 0.02 1.15 0.04 1.12
assay. (8.08) (0.58) (33.17) (1.18) (32.30)
• Results should be used in conjunction with other data; e.g., Tanner Stage II 24 0.42 0.15 0.80 0.18 0.77
symptoms, results of other tests, and clinical impressions. (12.11) (4.33) (23.07) (5.05) (22.21)
• If the testosterone results are inconsistent with clinical evidence, Tanner Stage III 24 0.89 0.24 1.49 0.26 1.43
additional testing is recommended. (25.67) (6.92) (42.97) (7.43) (41.24)
• Specimens from patients who have received preparations of Tanner Stage IV 19 0.94 0.53 1.86 0.53 1.86
mouse monoclonal antibodies for diagnosis or therapy may (27.11) (15.29) (53.64) (15.29) (53.64)
contain human anti-mouse antibodies (HAMA). Such specimens Tanner Stage V 24 1.31 0.52 3.55 0.59 3.43
may show either falsely elevated or depressed values when (37.78) (15.00) (102.38) (17.02) (98.78)
tested with assay kits such as ARCHITECT 2nd Generation
A third study was conducted testing a minimum of 120 samples
Testosterone that employ mouse monoclonal antibodies.
from individuals in the following categories: normal males 21-49
Additional information may be required for diagnosis.11, 12
years of age, normal males ≥ 50 years of age, premenopausal
• Heterophilic antibodies in human serum can react with reagent normal females 21-49 years of age, and postmenopausal normal
immunoglobulins, interfering with in vitro immunoassays. Patients females ≥ 50 years of age not on hormone replacement therapy.
routinely exposed to animals or to animal serum products can The Free Testosterone Index (% FTI) or Free Androgen Index (% FAI)
be prone to this interference, and anomalous values may be correlates with the value of free testosterone. Therefore, in addition
observed. Additional information may be required for diagnosis.13 to ARCHITECT 2nd Generation Testosterone (List Number 2P13), all
• A strong interaction with 19-nortestosterone (Nandrolone) was samples were tested with ARCHITECT Sex Hormone Binding Globulin
found. Do not use samples from patients receiving Nandrolone (ARCHITECT SHBG, List Number 8K26). The % FTI or % FAI was
treatment. calculated on a molar/molar basis. These samples gave values for
ll
EXPECTED VALUES the different groups summarized in the following table*.
The expected ranges for the ARCHITECT 2nd Generation
Testosterone assay were obtained from testing a minimum of
120 samples from apparently healthy individuals in the following
categories: normal males, 21-49 years of age with an intact
reproductive system, and normal females 21-49 years of age.
Additional samples were tested from apparently healthy males
and females (≥ 50 years of age). The data are summarized in the
following table.

5
 SHBG and Total Testosterone Within -
Testosterone nmol/L Mean Laboratory
SHBG nmol/L (ng/dL) Reagent nmol/L Within Run (Total)
2.5th 97.5th 2.5th 97.5th Sample Instrument Lot n (ng/dL) SD %CV SD %CV
Category n Median percentile percentile Median percentile percentile Control 1 1 80 0.34 0.016 4.6 0.017 4.8
Males 163 31.1 16.2 68.5 15.33 8.76 27.85 Level 1 (9.88) (0.456) (0.477)
(21-49 years of (442.07) (252.73) (803.24) 2 80 0.32 0.012 3.9 0.016 5.1
age) (9.24) (0.358) (0.472)
Males 144 35.3 13.7 69.9 14.42 8.58 23.37 2 1 80 0.33 0.014 4.1 0.015 4.6
( ≥50 years of (415.85) (247.50) (674.13) (9.56) (0.390) (0.439)
age) 2 80 0.31 0.016 5.1 0.016 5.2
Females 174 48.6 14.7 122.5 1.05 0.52 1.72 (9.02) (0.459) (0.468)
(Premenopausal, (30.43) (14.92) (49.56) Control 1 1 80 2.64 0.081 3.1 0.095 3.6
21-49 years of Level 2 (76.07) (2.339) (2.734)
age)
2 80 2.50 0.079 3.2 0.082 3.3
Females 175 49.9 16.7 124.4 0.76 0.46 1.18
(72.07) (2.277) (2.361)
(Postmenopausal, (21.83) (13.34) (33.90)
≥50 years of age) 2 1 80 2.57 0.086 3.4 0.092 3.6
(74.24) (2.492) (2.661)
Free Testosterone Index or Free Androgen Index
2 80 2.53 0.061 2.4 0.093 3.7
FTI or FAI (%) a (72.86) (1.769) (2.684)
2.5th 97.5th Control 1 1 80 7.92 0.167 2.1 0.208 2.6
Category n Median percentile percentile Level 3 (228.38) (4.811) (5.993)
Males 163 46.6 24.5 113.3 2 80 7.86 0.161 2.0 0.208 2.7
(21-49 years of age) (226.67) (4.643) (6.008)
Males 144 40.7 19.3 118.4 2 1 80 7.88 0.199 2.5 0.260 3.3
( ≥50 years of age) (227.22) (5.732) (7.496)
Females 174 2.0 0.7 8.7 2 80 7.97 0.191 2.4 0.222 2.8
(Premenopausal, (229.95) (5.504) (6.391)
21-49 years of age)
Panel 1 1 80 2.16 0.079 3.7 0.082 3.8
Females 175 1.5 0.5 4.7 (62.35) (2.287) (2.379)
(Postmenopausal,
2 80 2.11 0.045 2.1 0.062 3.0
≥50 years of age)
(60.72) (1.285) (1.796)
a FTI or FAI (%) = ARCHITECT 2nd Generation Testosterone Value (nmol/L) 2 1 80 2.13 0.065 3.1 0.076 3.6
x 100
ARCHITECT SHBG (nmol/L) (61.30) (1.888) (2.184)
2 80 2.13 0.054 2.5 0.070 3.3
* Representative performance data are shown. Results obtained at
(61.53) (1.558) (2.028)
individual laboratories may vary.
It is recommended that each laboratory establish its own reference Linearity
range that is appropriate for the laboratory’s patient population (i.e., A study was performed based on guidance from the NCCLS
a normal range that reflects the type of specimen and demographic document EP6-A.15 Using an absolute deviation from linearity of
variables such as age and sex, as applicable). 0.125 nmol/L for samples with concentrations of ≤ 0.5 nmol/L, and
ll
SPECIFIC PERFORMANCE CHARACTERISTICS 10% for samples with concentrations > 0.5 nmol/L, a linear range of
Data in the SPECIFIC PERFORMANCE CHARACTERISTICS section 0.13 – 64.57 nmol/L (3.82 – 1862.27 ng/dL) was demonstrated for
were generated using the ARCHITECT i 2000SR System. Assay the ARCHITECT 2nd Generation Testosterone assay.
results obtained in individual laboratories may vary from data Sensitivity
presented. Limit of Quantitation
Precision The ARCHITECT 2nd Generation Testosterone assay is designed
The ARCHITECT 2nd Generation Testosterone assay is designed to to have a Limit of Quantitation (LoQ) of ≤ 0.15 nmol/L, where the
have a within-laboratory (total) imprecision of ≤ 10% CV for samples LoQ is defined as the lowest analyte concentration that yields an
with testosterone concentrations ≥ 0.5 nmol/L to 35 nmol/L. estimated total error less than the total allowable error (TEa), where,
Within-Laboratory Precision the TEa is:
A study was performed based on guidance from the National • TEa = 0.125 nmol/L for samples with concentrations
Committee for Clinical Laboratory Standards (NCCLS) document ≤ 0.5 nmol/L
EP5-A2.14 Testing was conducted using two instruments, two • TEa = 25% for samples with concentrations > 0.5 nmol/L
ARCHITECT 2nd Generation Testosterone Reagent Kit lots, and one The LoQ was determined based on guidance from NCCLS document
lot each of ARCHITECT 2nd Generation Testosterone Calibrators and EP17-A.16 The observed LoQ for the ARCHITECT 2nd Generation
Controls. Three levels of controls and 1 human serum panel were Testosterone assay was 0.08 nmol/L (2.30 ng/dL).
assayed with a minimum of 2 replicates at 2 separate times per day Limit of Blank and Limit of Detection
for 20 different days. The data are summarized in the following table. In the same study, the Limit of Blank (LoB) and Limit of Detection
(LoD) were determined. The LoB was 0.03 nmol/L (0.87 ng/dL) and
the LoD was 0.05 nmol/L (1.44 ng/dL).
Interference
Potentially Interfering Endogenous Substances
Potential interference in the ARCHITECT 2nd Generation Testosterone
assay from hemoglobin, bilirubin, triglycerides, protein and biotin
was evaluated to be ≤ 10%. Interference was demonstrated by a

6
study based on guidance from the Clinical and Laboratory Standards Testosterone Concentration
Institute (CLSI, formerly NCCLS) protocol EP7-A2.17 The data are Test
Compound 2.4 nmol/L 10 nmol/L
summarized in the following table.
Test Compound Conc.a % Cross- % Cross-
% Interference (Drugs) nmol/L Conc. Diff.b Reactivityc Conc. Diff.b Reactivityc
Minimum
Potentially Interfering Interferent Testosterone Concentration Testosterone 250 0.43 0.2 0.88 0.4
Endogenous Substance Concentration 7 nmol/L 24.3 nmol/L Enanthate
Bilirubin (unconjugated) 15 mg/dL 3.3 4.2 Triamcinolone 100 0.00 0.0 0.05 0.0
Bilirubin (conjugated) 15 mg/dL 1.5 4.1 Spironolactone 500 ng/mL -0.02 0.0 -0.10 0.0
Hemoglobin 100 mg/dL -1.4 -0.5 Testosterone Concentration
Total Protein 12 g/dL -4.6 -7.0 Test
Compound 2.4 nmol/L 10 nmol/L
Triglycerides 1,000 mg/dL -6.4 -4.4 Test Compound (Other Conc.a Conc. % Cross- Conc. % Cross-
Biotin 30 ng/mL -2.1 -0.8 Compounds) nmol/L Diff.b Reactivityc Diff.b Reactivityc
Aldosterone 500 -0.07 0.0 0.17 0.0
Potentially Interfering Drugs and Other Compounds
5α-androstan-3α,17β-diol 20 -0.01 -0.1 -0.10 -0.5
A study was performed based on guidance from the CLSI document
5α-androstan-3β,17β-diol 10 0.03 0.3 0.05 0.5
EP7-A2.17 Potentially interfering drugs and other compounds were
Androstanedione 1,000 0.07 0.0 -0.30 0.0
evaluated to determine whether testosterone concentrations were
Androstenediol 40 1.34 3.3 -2.16 -5.4
affected when using the ARCHITECT 2nd Generation Testosterone
assay. The data are summarized in the following table. Androstenedione 20 0.20 1.0 0.02 0.1
Androsterone 1,000 -0.10 0.0 0.02 0.0
NOTE: Test compound concentration is in nmol/L unless noted
otherwise. Androsterone glucuronide 1,000 -0.01 0.0 -0.01 0.0
Androsterone sulphate 1,000 0.00 0.0 -0.14 0.0
Testosterone Concentration
Test Corticosterone 5,000 0.10 0.0 0.22 0.0
Compound 2.4 nmol/L 10 nmol/L
Cortisol 10,000 0.12 0.0 0.18 0.0
Test Compound Conc.a % Cross- % Cross-
(Drugs) nmol/L Conc. Diff.b Reactivityc Conc. Diff.b Reactivityc Cortisone 1,000 -0.02 0.0 0.07 0.0
Buserelin 300 ng/mL -0.03 0.0 0.04 0.0 Desoxycorticosterone 1,000 0.37 0.0 0.55 0.1
Clomiphene Citrate 10,000 -0.08 0.0 0.01 0.0 DHEA 50 -0.01 0.0 -0.03 -0.1
Cyproterone acetate 2,000 -0.07 0.0 -0.16 0.0 DHEAS 50,000 1.34 0.0 0.05 0.0
Danazol 1,000 2.48 0.2 0.04 0.0 Dihydrotestosterone 40 0.13 0.3 -0.57 -1.4
11-deoxy-17- hy- 1,000 -0.01 0.0 0.12 0.0 Epiandrosterone 250 -0.02 0.0 -0.25 -0.1
droxycorticosterone Epitestosterone 100 0.05 0.0 0.01 0.0
Desoxycorticosterone 5,000 0.06 0.0 0.04 0.0 17α-estradiol 1,000 -0.08 0.0 -0.04 0.0
acetate 17β-estradiol 200 0.06 0.0 -0.31 -0.2
Dexamethasone 5,000 -0.01 0.0 0.04 0.0 17β-estradiol-3- 500 0.22 0.0 -0.68 -0.1
Diethylstilbestrol 2 μg/mL 0.01 0.0 -0.06 0.0 glucuronide
(DES) 17β-estradiol-3-sulphate 2,000 0.02 0.0 -0.12 0.0
17β-estradiol-17- 10,000 0.03 0.0 -0.24 0.0 Estriol 800 0.03 0.0 -0.34 0.0
propionate
Estriol 3-(β-D-glucuronide 500 0.02 0.0 -0.24 0.0
17β-estradiol-17- 10,000 0.05 0.0 0.00 0.0 sodium salt)
valerate
Estrone 500 -0.03 0.0 -0.03 0.0
Ethisterone 20 0.01 0.0 -0.37 -1.8
Ethynodiol diacetate 50 ng/mL 0.00 0.0 0.04 0.0
Flunisolide 1,000 -0.01 0.0 0.01 0.0
17α-ethynyl estradiol 1000 ng/mL 0.0 0.0 0.0 0.0
Fluoxymesterone 500 0.00 0.0 -0.34 -0.1
Etiocholan-3,17-dione 500 0.00 0.0 -0.04 0.0
Flutamide 250 ng/mL 0.03 0.0 -0.01 0.0
Etiocholan-3α,17β-diol 500 0.03 0.0 -0.34 -0.1
Goserelin acetate 10 ng/mL -0.05 -0.6 -0.05 -0.6
19-hydroxyandrostene- 100 0.07 0.1 -0.86 -0.9
Hydroxyflutamide 5 μg/mL 0.01 0.0 0.02 0.0 dione
Ketoconazole 20 μg/mL 0.02 0.0 0.06 0.0 16α-hydroxyestrone 400 0.01 0.0 -0.11 0.0
Leuprolide acetate 150 ng/mL 0.03 0.0 0.04 0.0 17α-hydroxypregnanolone 5,000 0.03 0.0 -0.01 0.0
Livial (Tibolone) 1,000 0.06 0.0 -0.21 0.0 17α-hydroxyprogesterone 5,000 0.05 0.0 -0.10 0.0
Lynestrenol 1,000 0.25 0.0 -0.58 -0.1 6β-hydroxytestosterone 5 0.42 8.4 -0.48 -9.6
Medroxyprogesterone 5,000 -0.06 0.0 -0.28 0.0 11β-hydroxytestosterone 5 1.53 30.6 0.60 12.0
Mestranol 1,000 0.02 0.0 0.24 0.0 11-ketotestosterone 5 0.07 1.4 -0.35 -6.9
(17α-ethynylestradiol ng/mL
17α-methyltestosterone 10 0.62 6.2 -0.40 -4.0
3 methyl ether)
Pregnanolone 2,000 -0.01 0.0 0.01 0.0
Nilutamide 25 μg/mL 0.03 0.0 -0.27 0.0
Progesterone 2,000 -0.03 0.0 0.04 0.0
Norethindrone 10 0.07 0.7 0.11 1.1
Norethindrone 500 0.05 0.0 -0.22 0.0 a Test compounds were tested at or above the listed concentration.
acetate b Conc. Diff. = Concentration Difference
Norgestrel 20 ng/mL 0.19 0.3 -0.61 -1.0
c % Cross-Reactivity = Concentration Difference
19-nortestosterone 30 106.11 353.7 98.65 328.8 x 100
(Nandrolone) Test Compound Concentration
Oxymethalone 100 0.04 0.0 0.16 0.2
Method Comparison
Prednisone 2000 0.03 0.0 -0.02 0.0
The ARCHITECT 2nd Generation Testosterone assay is designed
Stanazolol 400 0.17 0.0 -0.35 -0.1
to have a slope of 1.0 ± 0.2 and a correlation coefficient (r) of
Tamoxifen 1,000 0.01 0.0 -0.07 0.0 ≥ 0.95 for samples with testosterone concentrations ranging
Testosterone Acetate 250 4.80 1.9 2.33 0.9 from 0.15 nmol/L (LoQ) to 35 nmol/L when compared to Liquid
Chromatography - Tandem Mass Spectrometry (LCMS). A correlation
study was performed based on guidance from the NCCLS document

7
EP9-A218 using the Passing- Bablok regression method to compare ll
Key to Symbols
the ARCHITECT 2nd Generation Testosterone assay to the LCMS
Consult instructions for use
testosterone method using serum specimens (n = 198). The data are
summarized in the following table.
Manufacturer
Correlation of ARCHITECT 2nd Generation Testosterone to LCMS
Testosterone
Sufficient for
Concentration Range Correlation
(nmol/L) Coefficient (r) Intercept
ARCHITECT LCMS r 95% CLa (nmol/L) 95% CIb Slope 95% CIb Temperature limitation
0.38-32.53 0.34-32.79 0.964 0.952 0.05 (-0.06, 0.94 (0.92,
0.15) 0.96)
Use by/Expiration date
a 95% CL= Confidence Limit (Lower, One-sided)
b CI = Confidence Interval
Assay Specific Diluent
ll
BIBLIOGRAPHY Conjugate
1. Dunn JF, Nisula BC, and Rodbard D. Transport of Steroid Hormones:
Binding of 21 Endogenous Steroids to Both Testosterone-Binding Contains Sodium Azide. Contact
Globulin and Corticosteroid-Binding Globulin in Human Plasma. J. Clin with acids liberates very toxic
Endocrin. Metab. 1981;53:58-68. gas.
2. Vermeulen A, Verdonck L, Kaufman J, et al. A critical evaluation
of simple methods for the estimation of free testosterone in serum. Control Number
Journal of Endocrinology and Metabolism1999; 84:3666-3672. In Vitro Diagnostic Medical
3. Selby, C. Sex hormone binding globulin: origin, function and clinical
significance. Ann Clin Biochem. 1990;27:532-541.
Device
4. Pugeat M, Crave JC, Tourniare J, Forest MG. Clinical Utility of Sex Lot Number
Hormone Binding Globulin Measurement. Horm Res. 1996; 45 (3-
5):148-155. Microparticles
5. Braunstein GD. Androgen insufficiency in women: summary of critical Pre-Trigger Solution
issues. Fertility and Sterility 2002;77(4, suppl 4):S94–95.
6. Brambilla D, O’Donnell A, Matsumoto A, McKinlay J, et al. Produced for Abbott by
Intraindividual variation in levels of serum testosterone and other
reproductive and adrenal hormones in men. Clinical Endocrinology Product of United Kingdom
2007; 67:853-862. Reaction Vessels
7. US Department of Labor, Occupational Safety and Health
Administration, 29 CFR Part 1910.1030, Bloodborne pathogens. Reagent Lot
8. US Department of Health and Human Services. Biosafety in
Microbiological and Biomedical Laboratories. 5th ed. Washington, DC: List Number
US Government Printing Office; December 2009.
9. World Health Organization. Laboratory Biosafety Manual. 3rd ed. Replacement Caps
Geneva: World Health Organization; 2004. Sample Cups
10. Clinical and Laboratory Standards Institute (CLSI). Protection of
Laboratory Workers From Occupationally Acquired Infections; Approved Septum
Guideline—Fourth Edition. CLSI Document M29-A4. Wayne, PA: CLSI;
2014. Serial number
11. Primus FJ, Kelley EA, Hansen HJ, et al. “Sandwich”-type immunoassay Specimen Diluent
of carcinoembryonic antigen in patients receiving murine monoclonal
antibodies for diagnosis and therapy. Clin Chem 1988;34(2):261-264. Trigger Solution
12. Schroff RW, Foon KA, Beatty SM, et al. Human anti-murine
immunoglobulin responses in patients receiving monoclonal antibody Warning: May cause an allergic
therapy. Cancer Res 1985;45(2):879-885. reaction.
13. Boscato LM, Stuart MC. Heterophilic antibodies: a problem for all Wash Buffer
immunoassays. Clin Chem 1988;34(1):27-33.
14. National Committee for Clinical Laboratory Standards (NCCLS).
Evaluation of Precision Performance of Quantitative Measurement The following U.S. Patents are relevant to the ARCHITECT iSystem
Methods; Approved Guideline—Second Edition. NCCLS Document
EP5-A2. Wayne, PA: NCCLS; 2004. or its components. There are other such patents and patent
15. National Committee for Clinical Laboratory Standards (NCCLS). applications in the United States and worldwide.
Evaluation of the Linearity of Quantitative Measurement Procedures: 5 468 646 5 543 524 5 545 739
A Statistical Approach; Approved Guideline. NCCLS Document EP6-A.
Wayne, PA: NCCLS; 2003. 5 565 570 5 669 819 5 783 699
16. National Committee for Clinical Laboratory Standards (NCCLS).
Protocols for Determination of Limits of Detection and Limits of ARCHITECT and Chemiflex are trademarks of Abbott Laboratories
Quantitation; Approved Guideline. NCCLS Document EP17-A. Wayne, in various jurisdictions. All other trademarks are property of their
PA: NCCLS; 2004. respective owners.
17. Clinical and Laboratory Standards Institute (CLSI). Interference Testing
in Clinical Chemistry; Approved Guideline—Second Edition. CLSI Abbott GmbH & Co. KG
Document EP7-A2. Wayne, PA: CLSI; 2005. Max-Planck-Ring 2
18. National Committee for Clinical Laboratory Standards (NCCLS). Method 65205 Wiesbaden
Comparison and Bias Estimation Using Patient Samples; Approved Germany
Guideline—Second Edition. NCCLS Document EP9-A2. Wayne, PA: +49-6122-580
NCCLS; 2002.

Axis-Shield Diagnostics Ltd., Dundee, UK

Customer Service: Contact your local representative
or find country-specific contact information on
[Link]
Revised November 2016.
©2015, 2016 Abbott Laboratories