ADVA Centaur Testosterone II (TSTII), ADVA Centaur Testosterone II (TSTII) Master Curve Material, ADVIA Centaur SHBG, ADVIA Centaur SHBG Calibrator, ADVIA Centaur SHBG Master Curve Material
Device Facts
| Record ID | K151986 |
|---|---|
| Device Name | ADVA Centaur Testosterone II (TSTII), ADVA Centaur Testosterone II (TSTII) Master Curve Material, ADVIA Centaur SHBG, ADVIA Centaur SHBG Calibrator, ADVIA Centaur SHBG Master Curve Material |
| Applicant | Siemens Healthcare Diagnostics, Inc. |
| Product Code | CDZ · Clinical Chemistry |
| Decision Date | Jun 29, 2016 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1680 |
| Device Class | Class 1 |
| Attributes | Real-World Evidence, Pediatric |
Real-World Evidence
| Submission | Device | Sponsor | RWD Sources | RWE Use Summary | Key Tags |
|---|---|---|---|---|---|
| K151986 · Jun 29, 2016 | ADVA Centaur Testosterone II (TSTII), ADVA Centaur Testosterone II (TSTII) Master Curve Material, ADVIA Centaur SHBG, ADVIA Centaur SHBG Calibrator, ADVIA Centaur SHBG Master Curve Material | Siemens Healthcare Diagnostics, Inc. | Clinical laboratory reference interval study cohorts (apparently healthy pediatric and adult populations) | Retrospective clinical data from healthy pediatric and adult subjects were used to establish reference intervals for the ADVIA Centaur TSTII and SHBG assays in accordance with CLSI EP28-A3c guidelines. | Reference Intervals; Clinical Cohort; Pediatric Population |
Clinical Evidence
| Study Design | Population | Comparator | Key Endpoints |
|---|---|---|---|
| Reference Interval Establishment; Retrospective analysis of clinical samples from healthy populations | Apparently healthy pediatric (age 2-21) and adult (age 21-89) subjects; Sample Size: Multiple cohorts ranging from 20 to 167 subjects per group | Not applicable for this study | Establishment of reference intervals for Testosterone and SHBG |
Indications for Use
The ADVIA Centaur Testosterone II (TSTII) assay is for in vitro diagnostic use in the quantitative determination of total testosterone (bound and unbound) in human serum and plasma using the ADVIA Centaur XP system. Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females, hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes. The ADVIA Centaur Testosterone II (TSTII) Master Curve Material is for in vitro diagnostic use in the verification of calibration and reportable range of the ADVIA Centaur Testosterone II (TSTII) assay. The ADVIA Centaur SHBG assay is an in vitro diagnostic immunoassay for the quantitative determination of sex hormone-binding globulin (SHBG) in human serum and plasma using the ADVIA Centaur XP system. The ADVIA Centaur SHBG assay is intended for use as an aid in the diagnosis of androgen disorders. The ADVIA Centaur SHBG Calibrator is for in vitro diagnostic use in calibrating the ADVIA Centaur SHBG assay on the ADVIA Centaur systems. The ADVIA Centaur SHBG Master Curve Material is for in vitro diagnostic use in the verification of calibration and reportable range of the ADVIA Centaur SHBG assay.
Device Story
ADVIA Centaur TSTII and SHBG assays are quantitative immunoassays for use on the ADVIA Centaur XP system. TSTII is a competitive chemiluminescent immunoassay; SHBG is a sandwich chemiluminescent immunoassay. TSTII uses a releasing agent to dissociate testosterone from endogenous binding proteins. Both assays utilize magnetic latex particles and acridinium ester-labeled reagents. The system detects relative light units (RLUs) proportional (SHBG) or inversely proportional (TSTII) to analyte concentration. Results are calculated via master curves adjusted by high/low calibrators. Used in clinical laboratories by trained personnel. Output aids clinicians in diagnosing androgen-related disorders. Benefits include standardized, automated measurement of hormone levels to support clinical decision-making.
Clinical Evidence
Bench testing only. Precision study (20 days, 80 replicates) showed total %CV 5.3-12.6% for TSTII. Linearity confirmed 7.0-1500 ng/dL (TSTII) and 1.6-180 nmol/L (SHBG). Method comparison against CDC ID-LC-MS/MS (n=128) yielded slope 0.971, r=0.982. Pediatric samples (n=61) showed r=0.99. Interference testing evaluated hemoglobin, bilirubin, triglycerides, and biotin; limitations noted for high-dose biotin and specific bilirubin levels. HAMA interference testing showed mean recovery 94.55-98.69%.
Technological Characteristics
Competitive (TSTII) and sandwich (SHBG) chemiluminescent immunoassays. Components: acridinium ester-labeled haptens/antibodies, streptavidin-coated magnetic latex particles. Energy source: ADVIA Centaur XP system. Connectivity: Standalone analyzer. Calibration: 2-point. Traceability: TSTII to CDC ID-LC-MS/MS; SHBG to WHO 2nd IS (08/226). Software: Embedded firmware for system control and data processing.
Indications for Use
Indicated for quantitative determination of total testosterone and SHBG in human serum/plasma to aid in diagnosis/treatment of androgen disorders (e.g., hypogonadism, delayed/precocious puberty, impotence, hirsutism, virilization) in male and female patients.
Regulatory Classification
Identification
A testosterone test system is a device intended to measure testosterone (a male sex hormone) in serum, plasma, and urine. Measurement of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.
Predicate Devices
- Roche Diagnostics Elecsys Testosterone II (k093421)
- Siemens Healthcare Diagnostics ADVIA Centaur Testosterone (TSTO) Master Curve Material (k140505)
- Siemens Healthcare Diagnostics ADVIA Centaur SHBG Assay (k091867)
- Siemens Healthcare Diagnostics ADVIA Centaur SHBG Calibrator (k091867)
- Siemens Healthcare Diagnostics ADVIA Centaur SHBG Master Curve Material (k091867)
Submission Summary (Full Text)
{0}
1
# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE
A. 510(k) Number:
k151986
B. Purpose for Submission:
New device (ADVIA Centaur Testosterone II) and modification to the traceability and value assignment procedure to the ADVIA Centaur SHBG Assay cleared in k091867
C. Measurand:
Testosterone (TST) and Sex Hormone Binding Globulin (SHBG)
D. Type of Test:
Quantitative, competitive immunoassay (for Testosterone) and sandwich immunoassay (for SHBG)
E. Applicant:
Siemens Healthcare Diagnostics Inc.
F. Proprietary and Established Names:
ADVIA Centaur Testosterone II (TSTII), ADVIA Centaur Testosterone II (TSTII) Master Curve Material, ADVIA Centaur SHBG, ADVIA Centaur SHBG Calibrator, and ADVIA Centaur SHBG Master Curve Material
G. Regulatory Information:
1. Regulation section:
21 CFR 862.1680 Testosterone test system
21 CFR 862.1150 Calibrator
21 CFR 862.1660 Quality control material
2. Classification:
Class I, reserved (assays and master curve materials)
Class II (calibrator)
3. Product code:
CDZ Radioimmunoassay, testosterones and dihydrotestosterone
JIT Calibrator, secondary
JJX Single (specified) analyte controls
4. Panel:
Clinical Chemistry (75)
{1}
H. Intended Use:
1. Intended use(s):
See indication(s) for use below
2. Indication(s) for use:
The ADVIA Centaur Testosterone II (TSTII) assay is for in vitro diagnostic use in the quantitative determination of total testosterone (bound and unbound) in human serum and plasma using the ADVIA Centaur XP system. Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females, hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes.
The ADVIA Centaur Testosterone II (TSTII) Master Curve Material is for in vitro diagnostic use in the verification of calibration and reportable range of the ADVIA Centaur Testosterone II (TSTII) assay.
The ADVIA Centaur SHBG assay is an in vitro diagnostic immunoassay for the quantitative determination of sex hormone-binding globulin (SHBG) in human serum and plasma using the ADVIA Centaur XP system. The ADVIA Centaur SHBG assay is intended for use as an aid in the diagnosis of androgen disorders.
ADVIA Centaur SHBG Calibrator: For in vitro diagnostic use in calibrating the ADVIA Centaur SHBG assay on the ADVIA Centaur systems.
The ADVIA Centaur SHBG Master Curve Material is for in vitro diagnostic use in the verification of calibration and reportable range of the ADVIA Centaur SHBG assay.
3. Special conditions for use statement(s):
- For prescription use only
- For in vitro diagnostic use
4. Special instrument requirements:
All studies were performed using the ADVIA Centaur XP
{2}
I. Device Description:
| ADVIA Centaur TSTII Assay Components | | |
| --- | --- | --- |
| Component | Volume | Ingredients |
| ADVIA Centaur TSTII Primary Reagent ReadyPack (included in assay kit) | | |
| ADVIA Centaur TSTII Lite Reagent | 10.0 mL/pack | Acridinium ester-labeled hapten (36 μg/mL) in buffered saline with preservatives |
| ADVIA Centaur TSTII Solid Phase Reagent | 17.0 mL/pack | Streptavidin-coated latex particles (0.33 g/L) in buffered saline with preservatives |
| ADVIA Centaur TSTII Ancillary Reagent ReadyPack (included in assay kit) | | |
| ADVIA Centaur TSTII Releasing Agent | 10.0 mL/pack | Steroid releasing agent (0.4 μg/mL) and biotinylated sheep monoclonal anti-testosterone antibody (27 μg/L) in buffered saline and preservatives |
| ADVIA Centaur TSTII Calibrator (included in assay kit) | | |
| ADVIA Centaur TSTII Low and High Calibrators | 2.0 mL/vial | After reconstitution, low or high levels of USP-grade estradiol, testosterone, cortisol, and progesterone spiked in charcoal-stripped, defibrinated human plasma with sodium azide (0.1%) and preservatives |
| ADVIA Centaur TSTII Master Curve Material (sold separately) | | |
| ADVIA Centaur TSTII MCM1 | 1.0 mL/vial | Lyophilized human plasma with sodium azide (0.1% after reconstitution) and preservatives |
| ADVIA Centaur TSTII MCM2-7 | 1.0 mL/vial | Various levels of testosterone in lyophilized human plasma with sodium azide (0.1% after reconstitution) and preservatives |
3
{3}
| Summary of Ingredients of the ADVIA Centaur SHBG Assay Components | | |
| --- | --- | --- |
| Component | Volume | Ingredients |
| ADVIA Centaur SHBG Primary Reagent ReadyPack (included in assay kit) | | |
| ADVIA Centaur SHBG Lite Reagent | 3.0 mL/pack | Mouse monoclonal anti-SHBG antibody (~130 ng/mL) labeled with acridinium ester in HEPES-buffered saline with bovine serum albumin, mouse serum, sodium azide (< 0.1%), surfactant, and preservatives |
| ADVIA Centaur SHBG Solid Phase Reagent | 11.0 mL/pack | Streptavidin coupled to latex paramagnetic particles (~150 μg/mL) in HEPES buffered saline with bovine serum albumin, mouse serum, sodium azide (< 0.1%), surfactant, and preservatives |
| ADVIA Centaur SHBG Ancillary Well Reagent | 3.0 mL/pack | Biotinylated monoclonal mouse anti-SHBG antibody (~6 μg/mL) in HEPES buffered saline with bovine serum albumin, mouse serum, sodium azide (< 0.1%), surfactant, and preservatives |
| ADVIA Centaur SHBG Calibrator (sold separately) | | |
| ADVIA Centaur SHBG Low and High Calibrators | 2.0 mL/vial | After reconstitution, low or high levels of SHBG in equine serum, detergents and preservatives |
| ADVIA Centaur SHBG Master Curve Material (sold separately) | | |
| ADVIA Centaur SHBG MCM1-5 | 1.0 mL/vial | Various levels of SHBG in equine serum with detergents and preservatives |
All human source materials were tested by FDA approved methods and found to be non-reactive for HIV $\frac{1}{2}$ , HBsAg, and HCV.
# J. Substantial Equivalence Information:
1. Predicate Device name(s):
Roche Diagnostics Elecsys Testosterone II (k093421)
Siemens Healthcare Diagnostics ADVIA Centaur Testosterone (TSTO) Master Curve Material (k140505)
Siemens Healthcare Diagnostics ADVIA Centaur SHBG Assay (k091867)
Siemens Healthcare Diagnostics ADVIA Centaur SHBG Calibrator (k091867)
Siemens Healthcare Diagnostics ADVIA Centaur SHBG Master Curve Material (k091867)
{4}
2. Predicate 510(k) number(s):
k093421
k140505
k091867
3. Comparison with predicate:
Testosterone assay
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Candidate Device ADVIA Centaur Testosterone TSTII | Predicate Device Elecsys Testosterone II (k093421) |
| Intended Use | For in vitro diagnostic use in the quantitative determination of total testosterone (bound and unbound) in serum. Measurements of testosterone are used in the diagnosis and treatment of disorders involving the male sex hormones (androgens), including primary and secondary hypogonadism, delayed or precocious puberty, impotence in males and, in females hirsutism (excessive hair) and virilization (masculinization) due to tumors, polycystic ovaries, and adrenogenital syndromes. | Same |
| Calibration | 2 Point calibration | Same |
| Reportable range | 7.0-1500 ng/dL | 2.5-1500 ng/dL |
| Methodology | Chemiluminescence | Electrochemiluminescence |
| Traceability | ID-LC-MS/MS (CDC reference measurement) | ID-GC/MS |
{5}
Testosterone Master Curve Material
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Candidate Device ADVIA Centaur TST II Master Curve Material | Predicate Device ADVIA Centaur Testosterone Master Curve Material (k140505) |
| Intended Use | The ADVIA Centaur TST II Master Curve Material is for the in vitro diagnostic use in the verification of calibration and reportable range of the ADVIA Centaur Testosterone II assay | Same |
| Matrix | Lyophilized human plasma | Same |
| Levels | 7 | Same |
| Target values | Level 1: 0.00 ng/dL
Level 2: 50.0 ng/dL
Level 3: 100.0 ng/dL
Level 4: 500.0 ng/dL
Level 5: 750.0 ng/dL
Level 6: 1000.0 ng/dL
Level 7: 1600.0 ng/dL | Same |
| Value assignment Process | Full curve testing | Two-point nested testing. |
SHBG assay
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Candidate Device ADVIA Centaur SHBG Assay | Predicate Device ADVIA Centaur SHBG Assay (k091867) |
| Intended Use | An in vitro diagnostic immunoassay for the quantitative determination of sex hormone-binding globulin (SHBG) in human serum and plasma. The ADVIA Centaur SHBG assay is intended for use as an aid in the diagnosis of androgen disorders. | Same |
| Calibration | 2 Point calibration | Same |
| Reportable range | 1.60–180 nmol/L | Same |
| Methodology | Chemiluminescence | Same |
| Traceability | WHO 2nd International Standard (08/226) | WHO 1st International Standard (95/560) |
{6}
SHBG Calibrators
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Candidate Device
ADVIA Centaur SHBG Calibrators | Predicate Device
ADVIA Centaur SHBG Calibrators
(k091867) |
| Intended Use | For in vitro diagnostic use in calibrating the ADVIA Centaur SHBG assay on the ADVIA Centaur systems. | Same |
| Matrix | Equine serum | Same |
| Levels | Low and high | Same |
| Concentration | Calibrator 1:10 nmol/L SHBG;
Calibrator 2:160 nmol/L SHBG. | Same |
| Traceability | WHO 2nd International Standard (08/226) | WHO 1st International Standard (95/560) |
| Value assignment process | Full curve testing | Two-point nested testing |
SHBG Master Curve Material
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Candidate Device
ADVIA Centaur SHBG Master Curve Material | Predicate Device
ADVIA Centaur SHBG Master Curve Material
(k091867) |
| Intended Use | The ADVIA Centaur SHBG Master Curve Material is for the in vitro diagnostic use in the verification of calibration and reportable range of the ADVIA Centaur SHBG assay | Same |
| Matrix | Equine serum | Same |
| Levels | 5 levels at approximately 0, 10, 50, 100, 200 nmol/L | Same |
| Value assignment process | Two point nested testing | Same |
| Traceability | WHO 2nd International Standard (08/226) | WHO 1st International Standard (95/560) |
{7}
K. Standard/Guidance Document Referenced (if applicable):
- CLSI EP05-A2: Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline-Second Edition
- CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline-First Edition
- CLSI EP07-A2: Interference Testing in Clinical Chemistry; Approved Guideline; Approved Guideline-Second Edition
- CLSI EP9-A3: Measurement Procedure Comparison And Bias Estimation Using Patient Samples; Approved Guideline - Third Edition
- CLSI EP17-A2: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline-Second Edition
- CLSI EP28-A3c: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline-Third Edition
L. Test Principle:
The ADVIA Centaur TSTII assay is a competitive immunoassay using direct chemiluminescent technology. Testosterone in the sample competes with acridinium ester-labeled hapten in the Lite Reagent for binding with the anti-testosterone sheep monoclonal antibody. The assay uses ADVIA Centaur Testosterone releasing agent to release bound testosterone from the endogenous binding proteins in the sample. An inverse relationship exists between the amount of testosterone present in the sample and the amount of relative light units (RLUs) detected by the system. Results are calculated from a master curve adjusted by using high and low calibrators.
The ADVIA Centaur SHBG assay is a sandwich immunoassay that uses three reagent components: Solid Phase, Ancillary Well Reagent, and Lite Reagent. Solid Phase is streptavidin-coupled magnetic latex particles, Ancillary Well Reagent contains biotinylated F(ab)2 monoclonal antibody to SHBG, and Lite Reagent consists of a second F(ab)2 monoclonal antibody to SHBG labeled with acridinium ester (SHBG-AE). The amount of SHBG present in the patient sample is proportional to the amount of relative light units (RLUs) detected by the system. Results are calculated from a master curve adjusted by using high and low calibrators.
M. Performance Characteristics (if/when applicable):
1. Analytical performance:
For ADVIA Centaur SHBG assay, sponsor has conducted a precision study and detection limits study to verify that the traceability change of the new assay do not have any significant impact on the performance of the precision and detection limits results of the unmodified assay (originally cleared SHBG assay in k091867).
a. Precision/Reproducibility:
{8}
# Testosterone:
The precision study was performed for twenty (20) days in accordance with CLSI EP5-A3 for the ADVIA Centaur TSTII assay on 3 different ADVIA Centaur XP instruments using 3 different lots of reagent. Each run was evaluated with a single replicate from two separate aliquots of 3 levels of controls (Bio-Rad Lyphochek), 5 levels of serum patient pools spiked with testosterone (Medical Decision Pools; MDPs), and one low native female patient specimen. Samples were tested twice a day in replicates of 2, for 20 days for a total of 80 replicates per sample. Each lot produced similar precision data. Data analysis from one representative lot is show below:
| | | | Within-run | | Total | |
| --- | --- | --- | --- | --- | --- | --- |
| Sample | N | Mean (ng/dL) | SD | %CV | SD | %CV |
| Serum from female | 80 | 10.58 | 0.90 | 8.5 | 1.33 | 12.6 |
| Serum pool 1 | 80 | 26.45 | 1.41 | 5.3 | 2.07 | 7.8 |
| Serum pool 2 | 80 | 81.78 | 2.49 | 3.0 | 4.29 | 5.3 |
| Serum pool 3 | 80 | 311.97 | 12.37 | 4.0 | 16.54 | 5.3 |
| Serum pool 4 | 80 | 768.74 | 23.81 | 3.1 | 41.71 | 5.4 |
| Serum pool 5 | 80 | 1127.07 | 54.15 | 4.9 | 77.67 | 6.9 |
| Control 1 | 80 | 226.34 | 7.11 | 3.1 | 10.19 | 4.5 |
| Control 2 | 80 | 578.19 | 19.28 | 3.3 | 34.53 | 6.0 |
| Control 3 | 80 | 735.72 | 24.61 | 3.3 | 44.43 | 6.0 |
SHBG: see previous performance in k091867
# b. Linearity/assay reportable range:
Testosterone:
A linearity study was performed by preparing intermediate concentrations of testosterone by diluting a high concentration testosterone human serum sample with a low concentration human serum sample for a total of 11 samples equally distributed
{9}
over the entire claimed measuring range (7.0 to 1500 ng/dL). The samples were run in duplicate. The test results ranged from 6.72 to 1654 ng/dL. The deviation of the ADVIA testosterone results from the expected linear fit was analyzed. The linear regression formula obtained from the analysis of the results was as follows: y = 0.92x - 0.22 ng/dL.
The results from the linearity study support an analytical measuring range from 7.0 ng/dL to 1500 ng/dL
## SHBG:
A linearity study was performed by preparing intermediate concentrations of SHBG by diluting a high concentration SHBG human serum sample with a SHBG-negative base-pool for a total of 11 samples equally distributed over the entire claimed measuring range (1.6 to 180.0 nmol/L). The samples were run in duplicate. The test results ranged from 1.16 to 195.95 nmol/L. The deviation of the ADVIA SHBG results from the expected linear fit was analyzed. The linear regression formula obtained from the analysis of the results was as follows: y = 0.984x + 0.245 ng/dL.
The results from the linearity study support an analytical measuring range from 1.6 to 180.0 nmol/L.
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
## Traceability:
### ADVIA Centaur TSTII Calibrators
The ADVIA Centaur TSTII Calibrators used for two-point calibration of the ADVIA Centaur TSTII assay are value assigned using standards which are traceable to isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS), (Center for Disease and Prevention(CDC) Hormone Standardization Program Testosterone Reference Measurement Procedure). The ID-LC-MS/MS is traceable to the primary testosterone standard National Measurement Institute (NMI) M914 reference material.
### ADVIA Centaur SHBG Calibrators
The ADVIA Centaur SHBG Calibrators used for two-point calibration of the ADVIA Centaur SHBG assay are traceable to the WHO 2nd International Standard for Sex Hormone Binding Globulin (08/226) from NIBSC.
## Calibrator value assignment:
### ADVIA Centaur TSTII Calibrators
The ADVIA Centaur TSTII calibrators are lot-locked to reagents (i.e. only one lot of
10
{10}
reagents is used during value assignment of each lot of calibrators). The calibrators are value assigned during the Master curve runs for a full curve value assignment. The value assignment consists of samples (Master curve standards, primary controls, low calibrator, and high calibrator) run in replicates of 6. The value assignment protocol requires 1 reagent lot (ADVIA Centaur TSTII reagent ReadyPacks with kit calibrators) using a minimum of 2 runs (1 run performed on 2 instruments), resulting in a total of 12 replicates per sample.
For each new lot of calibrators, the new calibrator values are calculated as the overall mean of the value assignment runs read off the standard curve. The calibrator target values and allowable ranges are presented below:
| Calibrator | Target value (ng/dL) | Range (ng/dL) |
| --- | --- | --- |
| Low | 27 | 17.4-36.6 |
| High | 1250 | 1162.6-1337.4 |
## ADVIA Centaur SHBG Calibrators:
The ADVIA Centaur SHBG Calibrators used for two-point calibration of the ADVIA Centaur SHBG assay are value assigned using standards prepared from the WHO 2nd International Standard for Sex Hormone Binding Globulin (08/226) from NIBSC. The in-process value assignment consists of samples (Primary standards, primary controls, low calibrator, and high calibrator) run in replicates of 6. The value assignment protocol requires 2 reagent lots (ADVIA Centaur SHBG reagent ReadyPacks) using a minimum of 2 runs (1 run performed with 2 reagent lots), resulting in a total of 12 replicates per sample. The calibrator target values and allowable ranges are presented below:
| Calibrator | Target value (nmol/L) | Range (nmol/L) |
| --- | --- | --- |
| Low | 10 | 8.0-12.0 |
| High | 140 | 119.0-160.0 |
## Value assignments of Testosterone and SHBG Master Curve Material:
The value assignment of the ADVIA Centaur TSTII Master Curve Material is verified by running three testosterone internal controls, and the results must meet internal specifications. The target values of the ADVIA Centaur TSTII MCM materials are listed below:
Level 1: 0.00 ng/dL
Level 2: 50.0 ng/dL
Level 3: 100.0 ng/dL
Level 4: 500.0 ng/dL
Level 5: 750.0 ng/dL
{11}
The value assignment of the ADVIA Centaur SHBG Master Curve Material is verified by running three levels of internal controls and three levels of external controls which must meet internal specifications. The target values of the ADVIA Centaur SHBG MCM materials listed are below:
Level 1: 0.00 nmol/L
Level 2: 10.0 nmol/L
Level 3: 50.0 nmol/L
Level 4: 100.0 nmol/L
Level 5: 200.0 nmol/L
Stability of SHBG and Testosterone Calibrators and Master Curve Material:
Stability study protocols and acceptance criteria were provided and found to be acceptable. The studies support the sponsor following stability claims:
ADVIA Centaur TSTII Calibrators:
- Shelf life: 6 months when stored at 2-8° C
- Open-vial: 14 days when stored at 2-8° C
- On board analyzer: 4 hours
ADVIA Centaur TSTII Master Curve Material:
- Unopened lyophilized: 20 months stored at 2-8° C
- Reconstituted Open-vial: 14 days stored at 2-8° C
- On board analyzer: 4 hours
ADVIA Centaur SHBG Calibrators:
- Unopened lyophilized: 39 weeks stored at 2-8° C
- Reconstituted Open-vial: 60 days stored at -20° C
ADVIA Centaur SHBG Master Curve Material:
- Unopened lyophilized: 39 weeks stored at 2-8° C
- Reconstituted Open-vial: use immediately and discard remaining material.
d. Detection limit:
Testosterone:
Limit of Blank (LoB)
Limit of Blank was determined for the ADVIA Centaur TSTII assay as described in CLSI Guideline EP17-A2. Testing was performed using 3 lots of ADVIA Centaur TSTII reagents and 5 ADVIA Centaur XP analyzers. For each lot/instrument combination, 80 determinations were obtained by testing 4 negative base-pools in duplicate (1 test each from two independent aliquots) twice a day for 5 days. The LoB was calculated non-parametrically following the recommendations in the CLSI guideline. The sponsor determined that the LoB of the assay is 1.09 ng/dL (the sponsor claims 2.5 ng/dL).
12
{12}
13
Limit of Detection (LoD)
Limit of Detection was determined for the ADVIA Centaur TSTII assay as described in CLSI Guideline EP17-A2. Testing was performed using 3 lots of ADVIA Centaur TSTII and two ADVIA Centaur XP instruments. For each lot/instrument combination, 80 determinations were obtained by testing 4 low analyte samples in duplicate (1 test each from two independent aliquots) twice a day for 5 days. The low analyte samples were prepared by pooling low level testosterone samples from different females. The LoD was determined following the parametric model described in the guideline. The sponsor determined that the LoD of the assay is 3.49 ng/dL (the sponsor claims 5.0 ng/dL).
Limit of Quantitation (LoQ)
Four low samples were prepared by pooling different female samples with low concentrations of testosterone. The 4 serum pools and a blank sample were tested in duplicate (1 test each from two independent aliquots). Testing was performed using 3 lots of ADVIA Centaur TSTII reagents, over 20 testing days, on five ADVIA Centaur XP instruments. The Within-lab %CV was plotted vs. the testosterone concentration to determine the LoQ (concentration at 20% CV). The LoQ was determined to be 7.00 ng/dL.
Detection Limits summary for ADVIA Centaur TSTII assay
| Limit of Blank | Limit of Detection | Limit of Quantitation |
| --- | --- | --- |
| 2.50 ng/dL | 5.00 ng/dL | 7.00 ng/dL |
The claimed measuring range of the ADVIA Centaur TSTII assay is 7.0-1500 ng/dL.
SHBG: see previous performance in k091867
e. Analytical specificity:
Testosterone:
i.) Potential cross-reactivity of 19 drugs and metabolites on the ADVIA Centaur TSTII assay were evaluated. Two sample pools of approximately 0 ng/dL and 300 ng/dL (serum patient pool spiked with testosterone to ~300 ng/dL testosterone) were spiked with each test compound so that the final test sample contained the compound at the required concentration. A control sample was prepared for each potential cross-reactant by spiking with the appropriate diluent at the same volume as the potential cross-reactant. Testing was performed in replicates of three (3) per sample with 2 reagent lots on an ADVIA Centaur XP instrument.
{13}
The mean of 3 replicates was used to calculate cross-reactivity using the following equation:
$$
\% \text {Cross - Reactivity} = 100 \times \frac {\left(\text {Dose of Test Sample - Dose of Control Sample}\right)}{\text {Concentration of Substance Added}}
$$
Cross-reactivity results from testing with 2 reagent lots are summarized below.
| Substance | Substance concentration (ng/mL) | Maximum % Cross Reactivity at 0 ng/dL TST | Maximum % Cross Reactivity at 300 ng/dL TST |
| --- | --- | --- | --- |
| 5-androstene-3β,17β-diol | 100,000 | 0.124 | -0.167 |
| Andostenedione | 10,000 | 1.397 | -0.886 |
| Androsterone | 100,000 | 0.003 | -0.026 |
| Canrenone | 18,000 | 0.000 | 0.066 |
| Canrenone | 50,000 | 0.001 | 0.005 |
| Canrenone | 100,000 | 0.001 | 0.008 |
| Corticosterone | 100,000 | 0.000 | 0.014 |
| Cortisol | 100,000 | 0.000 | 0.022 |
| Cyproterone | 10,000 | 0.128 | 0.054 |
| Danazol | 100,000 | 0.120 | -0.121 |
| 11-dexoycortisol | 100,000 | 0.002 | 0.002 |
| Dexamethasone | 100,000 | 0.000 | 0.006 |
| DHEA | 100,000 | 0.006 | -0.042 |
| DHEA-Sulfate | 5,000,000 | 0.001 | -0.001 |
| 5α-dihydro-testosterone | 10,000 | 0.754 | -1.410 |
| 17 β-estradiol | 100,000 | 0.021 | -0.267 |
| Esterone | 10,000 | 0.002 | -0.009 |
| Ethisterone | 100,000 | 0.446 | 0.408 |
| 11 β-hydroxytestosterone | 10,000 | 15.469 | 13.413 |
| 11-ketotestosterone | 100,000 | 1.766 | 1.583 |
| Nandrolone decanonate | 100,000 | * | * |
| Norgestrel | 100,000 | 0.145 | -0.073 |
| Oxymetholone | 10,000 | 0.008 | -0.576 |
| Prednisolone | 100,000 | 0.000 | 0.093 |
| Prednisone | 100,000 | 0.000 | 0.011 |
| Progesterone | 100,000 | 0.000 | 0.014 |
| Spironolactone | 8,000 | 0.005 | 0.105 |
| Spironolactone | 100,000 | 0.000 | 0.011 |
| Testosterone proprionate | 10,000 | 0.0356 | 2.946 |
| 7α-Thiomethyl spironolactone | 40,000 | 0.001 | 0.001 |
| 7α-Thiomethyl spironolactone | 100,000 | 0.000 | -0.002 |
{14}
*above assay measuring range
Based on the cross reactivity results, the sponsor included the following limitations in the labeling:
"A strong interaction with Nandrolone decanoate, $11\beta$ -hydroxytestosterone, and 11-keto-testosterone was found. Do not use samples from patients receiving these compounds."
"With the advent of new steroid-based medications (analogues) with similar chemical structures to testosterone, there is the possibility of cross-reactivity and falsely elevated results. For diagnostic purposes, the results should always be assessed in conjunction with the patient's medical history, clinical examination, and other findings. If the testosterone results are inconsistent with clinical evidence, additional testing is suggested to confirm the result."
# ii.) Interference Study:
The interference study was designed in accordance with CLSI EP7-A2 to evaluate the performance of the ADVIA Centaur TSTII assay in the presence of hemoglobin, intralipid (triglycerides), conjugated and unconjugated bilirubin, and biotin. A stock solution of each interferent was prepared and spiked into two serum patient pools with concentration approximately $30\mathrm{ng / dL}$ and $300\mathrm{ng / dL}$ testosterone. A control sample was prepared for each interfering substance by spiking with the appropriate diluent at the same volume as the interfering substance. All samples were run in triplicate with 2 reagent lots on an ADVIA Centaur XP instrument. The mean of 3 replicates was used to calculate percent interference using the following equation:
$\%$ Interference $= \frac{\text{spiked sample conc.} - \text{unspiked sample conc.}}{\text{unspiked sample conc.}}$ x100
In addition, a dose response with lower concentrations of endogenous substance was added to samples that did not meet acceptance criteria of $\leq \pm 10\%$ bias to determine the concentration at which interference was not significant. Sponsor defines significant interference as bias $>10\%$ between the tested sample and the control sample. The non-interference claims are listed in the following table.
| Substance | Highest Tested Concentration at which no significant interference (≤±10%) was observed |
| --- | --- |
| Hemoglobin | 500 mg/dL |
| Bilirubin (unconjugated) | 20 mg/dL |
| Bilirubin (conjugated) | 15 mg/dL |
| Triglycerides | 1000 mg/dL |
| Cholesterol | 500 mg/dL |
| Rheumatoid Factor | 200 IU/mL |
| Biotin | 30 ng/mL |
{15}
Based on the interference results, the sponsor included the following limitations in the labeling.
"Samples with conjugated bilirubin concentrations > 15 mg/dL will cause erroneous results. Specimens that contain 20 mg/dL of conjugated bilirubin demonstrate a -14.4% change in results."
"Samples with unconjugated bilirubin concentrations > 20 mg/dL will cause erroneous results. Specimens that contain 30 mg/dL of conjugated bilirubin demonstrate a 20% change in results"
"Specimens that contain 40 ng/mL of biotin demonstrate a 14.6% change in results." "In patients receiving therapy with high doses of biotin (> 5 mg/day), no sample should be taken until at least 8 hours after the last biotin administration."
iii.) Human anti-mouse antibodies (HAMA) interference: HAMA interference testing was performed by spiking 19 HAMA-positive serum samples with USP testosterone at concentrations of approximately 30 ng/dL and 300 ng/dL. The mean recovery for all samples spiked with 30 ng/dL testosterone was 98.69%. The mean recovery for all samples spiked with 300 ng/dL testosterone was 94.55%.
Sponsor has provided the limitations in their package insert:
"Heterophilic antibodies in human serum can react with reagent immunoglobulins, interfering with in vitro immunoassays. Patients routinely exposed to animals or to animal serum products can be prone to this interference and anomalous values may be observed. Additional information may be required for diagnosis."
SHBG: See previous performance in k091867
2. Comparison studies:
a. Method comparison with predicate device:
Testosterone:
The method comparison study was designed in accordance with CLSI EP9-A3 to compare the performance of the ADVIA Centaur TSTII assay to that of the recognized reference method (CDC HoSt Testosterone RMP ID-LC-MS/MS). The study evaluated a total of 128 individual male and female (adults) serum samples spanning the range of the assay. Of these, 88 samples were individual (non-pooled) human sera from single donors obtained by Siemens, with testosterone concentrations assigned by CDC HoSt Testosterone Reference Measurement Procedure (RMP) ID-LC-MS/MS. The remaining 40 samples were supplied directly by CDC, and were individual human serum obtained from individual blood bank donors, with testosterone concentrations assigned by CDC HoSt Testosterone RMP ID-LC-MS/MS. None of the samples were altered. All samples were run in singlicate on one
16
{16}
ADVIA Centaur XP system using one reagent TSTII lot over 2 days. The results for ADVIA Centaur TSTII versus CDC HoSt Testosterone RMP ID-LC-MS/MS were analyzed using Passing-Bablok regression analysis. The results are summarized below:
| N | 128 |
| --- | --- |
| Slope | 0.971 |
| Intercept | 1.944 |
| R | 0.982 |
| Test range | 8.61-1394 ng/dL |
An additional method comparison study was performed using 56 adult samples and 61 pediatric samples (with 27 female, age from 7 to 18 and 34 male, age from 9-18) against a commercially available Testosterone assay to support sponsor's claim that pediatric samples could be used with the candidate device. Weighted Deming regression was used for the data analysis. The combined total samples with range tested between 8.0 to $825\mathrm{ng / dL}$ resulted in $\mathrm{Y} = 1.01\mathrm{X} + 0.99$ , $\mathrm{r} = 0.99$ , $\mathrm{n} = 117$ . The pediatric samples with range tested between 8.0 to $732\mathrm{ng / dL}$ resulted in $\mathrm{Y} = 1.04\mathrm{X} + 0.22$ , $\mathrm{r} = 0.99$ , $\mathrm{n} = 61$ .
# SHBG:
A method comparison of the ADVIA Centaur SHBG assay to the Elecsys SHBG assay on the Roche Cobas e 411 analyzer was performed by testing 174 serum samples distributed over the assay range. In order cover the entire measuring range, 17 of the 174 (9.77%) samples were diluted. Data were analyzed using Weighted Deming regression. The results are summarized below:
| N | 174 |
| --- | --- |
| Slope | 0.99 |
| Intercept | 0.11 |
| R | 0.994 |
| Test range | 2.32 to 178.3 nmol/L |
# b. Matrix comparison:
# Testosterone:
Fifty matched sample sets spanning the assay range for serum, EDTA plasma, lithium heparin plasma, and sodium heparin plasma were run in singlicate using 2 lots of reagent. Five samples were altered in order to span the assay range. Data were analyzed by linear regression with results from plasma samples, plasma separator tubes (PSTs) and serum separator tubes (SSTs) were compared to results of serum samples in glass tubes. The results are summarized in the following table.
{17}
| Tube type | n | Slope | Intercept (ng/dL) | r |
| --- | --- | --- | --- | --- |
| Serum separator tube | 50 | 0.98 | 3.76 | 0.991 |
| Plasma separator tube | 50 | 0.94 | 3.56 | 0.993 |
| Dipotassium EDTA | 50 | 0.97 | -1.61 | 0.994 |
| Lithium heparin | 50 | 0.95 | 4.50 | 0.996 |
| Sodium heparin | 50 | 0.94 | 2.85 | 0.996 |
The matrix comparison study results support the use of serum separator serum, serum separator plasma, Li-heparin, Na-heparin, and K2-EDTA plasma with this assay.
SHBG: see previous performance in k091867
3. Clinical studies:
a. Clinical Sensitivity: Not applicable
b. Clinical specificity: Not applicable
c. Other clinical supportive data (when a. and b. are not applicable): Not applicable
4. Clinical cut-off: Not applicable
5. Expected values/Reference range:
Testosterone
A reference interval study was performed for the ADVIA Centaur Testosterone II assay. The sample groups tested consisted of males, $\geq 50$ years old (n=137); males between the ages of 21 to 49 years old (n=119); post-menopausal females, ages 48 to 89 years (n=127); and pre-menopausal females, ages 21 to 60 years (n=167). The central $95^{\text{th}}$ percentile range of each sample category was calculated by finding the $2.5^{\text{th}}$ ranked sample on the low end and the $97.5^{\text{th}}$ ranked sample on the high end. The follow
{18}
reference intervals were obtained for each category:
Adult males (21 to 49 yrs): 123.06-813.86 ng/dL
Adult males $(\geq 50$ yrs): 86.98-780.10 ng/dL
Pre-menopausal females: 9.01-47.94 ng/dL
Post-menopausal females: $< 7.0$ to $45.62\mathrm{ng / dL}$
A reference range study was performed on the pediatric population following CLSI EP28-A3c. Male and female pediatric patient samples were obtained spanning multiple age ranges and Tanner Stages. Each category of samples was analyzed separately. The reference intervals are presented in tables below for both Tanner Stages and age range.
Male Pediatric Reference Ranges According to Tanner Stage
| Tanner Stage | Gender | N | Lower range (ng/dL) | Upper range (ng/dL) |
| --- | --- | --- | --- | --- |
| Stage I | Male | 36 | <7 | 47.43 |
| Stage II | Male | 38 | <7 | 174.45 |
| Stage III | Male | 44 | 10.54 | 802.75 |
| Stage IV | Male | 44 | 63.89 | 736.20 |
| Stage V | Male | 42 | 55.69 | 897.40 |
Female Pediatric Reference Range According to Tanner Stage
| Tanner Stage | Gender | N | Lower range (ng/dL) | Upper range (ng/dL) |
| --- | --- | --- | --- | --- |
| Stage I | Female | 47 | <7 | 89.56 |
| Stage II | Female | 42 | <7 | 38.29 |
| Stage III | Female | 42 | <7 | 33.86 |
| Stage IV | Female | 40 | <7 | 38.72 |
| Stage V | Female | 42 | 10.83 | 50.08 |
Male Pediatric Reference Ranges According to Age
| Age | Gender | N | Lower range (ng/dL) | Upper range (ng/dL) |
| --- | --- | --- | --- | --- |
| 2-10 years | Male | 40 | <7 | 29.44 |
| 11 years | Male | 20 | <7 | 353.0 |
| 12 years | Male | 25 | <7 | 562.4 |
| 13 years | Male | 21 | 8.1 | 582.9 |
| 14 years | Male | 31 | 20.35 | 777.4 |
| 15 years | Male | 22 | 127.2 | 849.4 |
| 16-21 years | Male | 44 | 113.2 | 882.1 |
{19}
Female Pediatric Reference Ranges According to Age
| Age | Gender | N | Lower range (ng/dL) | Upper range (ng/dL) |
| --- | --- | --- | --- | --- |
| 2-10 years | Female | 40 | <7 | 117.8 |
| 11-15 years | Female | 130 | <7 | 38.9 |
| 16-21 years | Female | 36 | 15.1 | 42.4 |
Data Analysis: Each category of samples was analyzed separately. Due to the limited availability of pediatric samples, the sample analysis method varied depending on the number of samples in each subgroup. Please see the following descriptions of each subgroup reference intervals determination.
For groups of 120 or more subjects, the lower and upper reference limits were estimated as the $2.5^{\text{th}}$ and the $97.5^{\text{th}}$ percentiles of the distribution of test results using a non-parametric approach in accordance with the recommendation in CLSI guideline EP28-A3.
For groups of 40 to 119 subjects, the lower and upper reference limits were estimated as the $2.5^{\text{th}}$ and the $97.5^{\text{th}}$ percentiles of the distribution of test results using a robust measure of location and spread, as developed by Horn and Pesce.
For groups of less than 40 subjects, the lower and upper reference limits were estimated as the $5^{\text{th}}$ and the $95^{\text{th}}$ percentiles of the distribution of test results.
## SHBG
A reference interval study was performed for the ADVIA Centaur SHBG assay. The sample groups tested consisted of males, $>49$ years old $(n=137)$; males between the ages of 21 to 50 years old $(n=122)$; post-menopausal females, ages 48 to 89 years $(n=134)$; and pre-menopausal females, ages 21 to 60 years $(n=167)$. The central $95^{\text{th}}$ percentile range of each sample category was calculated by finding the $2.5^{\text{th}}$ ranked sample on the low end and the $97.5^{\text{th}}$ ranked sample on the high end. The follow reference intervals were obtained for each category:
- Adult males (21 to 49 yrs): 14.55 to 94.64 nmol/L
- Adult males ($\geq$50 yrs): 21.63 to 113.1 nmol/L
- Pre-menopausal females: 10.84 to $>180.0$ nmol/L
- Post-menopausal females: 23.15 to 159.1 nmol/L
{20}
# Free Androgen Index (FAI)
The free androgen Index reference range for the adult population ( $\geq 21$ yrs) was determined using the data collected from the reference interval studies performed from the ADVIA Centaur TSTII and ADVIA SHBG assays described above. The FAI was calculated for each data set using the following equation:
$$
\text {FAI} (\%) = \frac {\text {ADVIA Centaur TST II (nmol/L)}}{\text {ADVIA Centaur SHBG (nmol/L)}} \times 100
$$
The following reference intervals were obtained:
Free Androgen Index
| Gender | Age | N | Lower range (%) | Upper range (%) |
| --- | --- | --- | --- | --- |
| Male | 21-49 | 119 | 11.14 | 81.01 |
| Male | ≥50 | 137 | 8.99 | 46.51 |
| Female | 21-60 (premenopausal) | 167 | 0.27 | 7.64 |
| Female | 45-89 (postmenopausal) | 127 | 0.17 | 4.15 |
The sponsor states the following in the package insert labeling:
Consider these ranges as guidelines only. Each laboratory should determine its own reference ranges for the diagnostic evaluation of patient results.
# N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
# O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
