Elecsys Cortisol II, Cortisol II CalSet

K152227 · Roche Diagnostics · JFT · Apr 27, 2016 · Clinical Chemistry

Device Facts

• Record ID: K152227
• Device Name: Elecsys Cortisol II, Cortisol II CalSet
• Applicant: Roche Diagnostics
• Product Code: JFT · Clinical Chemistry
• Decision Date: Apr 27, 2016
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1205
• Device Class: Class 2

Intended Use

Immunoassay for the in vitro quantitative determination of cortisol in human serum, and plasma. The determination of cortisol is used for the recognition and treatment of functional disorders of the adrenal gland. The electrochemiluminescence immunoassay “ECLIA” is intended for use on Elecsys and cobas e immunoassay analyzers. Cortisol II CalSet is used for calibrating the quantitative Elecsys Cortisol II assay on the Elecsys and cobas e immunoassay analyzers

Device Story

Elecsys Cortisol II is an in vitro diagnostic immunoassay for quantitative cortisol measurement in human serum and plasma. It utilizes a competitive electrochemiluminescence immunoassay (ECLIA) principle on Elecsys and cobas e analyzers. The assay uses a monoclonal antibody specific to cortisol; endogenous cortisol is liberated from binding proteins via danazol and competes with a ruthenium-labeled exogenous cortisol derivative for binding sites on biotinylated antibodies. The device is operated by laboratory technicians in clinical settings. Results are calculated via a calibration curve generated by 2-point calibration and a master curve provided via reagent barcode. Healthcare providers use these results in conjunction with patient history and clinical examination to diagnose and manage adrenal gland disorders. The device benefits patients by providing accurate cortisol levels to support clinical decision-making for endocrine conditions.

Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP5-A2 (n=84, CVs 1.4-12.7%). Linearity confirmed 2.85-1898 nmol/L (R²=0.9995). LoB=1.0 nmol/L, LoD=1.5 nmol/L, LoQ=3.0 nmol/L. Interference testing showed no significant interference (≤10%) for common endogenous/exogenous substances. Cross-reactivity assessed for 16 compounds; limitations noted for prednisolone, 6-α-Methylprednisolone, prednisone, 11-deoxycortisol, and 21-deoxycortisol. Method comparison (n=536) vs predicate showed bias; comparison vs ID-GC-MS (n=208) showed good agreement (Slope 1.022, r=0.993). Reference ranges established from 300 healthy individuals.

Technological Characteristics

Quantitative competitive ECLIA. Reagents: biotinylated monoclonal anti-cortisol antibody (ovine), ruthenium-labeled cortisol derivative, streptavidin-coated microparticles, danazol, MES buffer. Platform: Elecsys/cobas e immunoassay analyzers. Standardization: IRMM/IFCC-451 panel via ID-GC/MS. Measuring range: 3.0–1750 nmol/L. Sample types: serum, Li-heparin plasma, K2/K3-EDTA plasma.

Indications for Use

Indicated for the in vitro quantitative determination of cortisol in human serum and plasma to assist in the recognition and treatment of functional disorders of the adrenal gland. Intended for use on Elecsys and cobas e immunoassay analyzers.

Regulatory Classification

Identification

A cortisol (hydrocortisone and hydroxycorticosterone) test system is a device intended to measure the cortisol hormones secreted by the adrenal gland in plasma and urine. Measurements of cortisol are used in the diagnosis and treatment of disorders of the adrenal gland.

Predicate Devices

• Roche Elecsys Cortisol and CalSet (k070788)

Submission Summary (Full Text)

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Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is circular, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. In the center of the circle is a stylized image of three human profiles facing to the right, stacked on top of each other.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

April 27, 2016

ROCHE DIAGNOSTICS KELLI TÜRNER REGULATORY AFFAIRS PRINCIPAL 9115 HAGUE ROAD INDIANAPOLIS IN 46250

Re: K15227

Trade/Device Name: Elecsys Cortisol II. Cortisol II CalSet Regulation Number: 21 CFR 862.1205 Regulation Name: Cortisol (hydrocortisone and hydroxycorticosterone) test system Regulatory Class: II Product Code: JFT, JIT Dated: March 24, 2016 Received: March 25, 2016

Dear Ms. Kelli Turner:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours.

# Katherine Serrano -S

For:

Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known) k152227

Device Name Elecsys Cortisol II

Indications for Use (Describe)

Immunoassay for the in vitro quantitative determination of cortisol in human serum, and plasma. The determination of cortisol is used for the recognition and treatment of functional disorders of the adrenal gland. The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys and cobas e immunoassay analyzers.

| Type of Use (Select one or both, as applicable) | |
|---------------------------------------------------------------------------------------------------------|---------------------------------------------|
| <div> <span> X </span> <span> Prescription Use (Part 21 CFR 801 Subpart D) </span> </div> | Over-The-Counter Use (21 CFR 801 Subpart C) |

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### DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

### Indications for Use

510(k) Number (if known)

### k152227

Device Name Cortisol II CalSet

Indications for Use (Describe)

Cortisol II CalSet is used for calibrating the quantitative Elecsys Cortisol II assay on the Elecsys and cobas e immunoassay analyzers.

Type of Use (Select one or both, as applicable)

> Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

### CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.*

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

> Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

Form Approved: OMB No. 0910-0120 Expiration Date: January 31, 2017 See PRA Statement below.

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# 510(k) Summary

# (k152227)

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

| Submitter Name | Roche Diagnostics |
|----------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Address | 9115 Hague Road<br>Indianapolis, IN 46250 |
| Contact | Kelli Turner<br>Phone: [phone]<br>FAX: [phone]<br>Email: [e-mail] |
| Date Prepared | 04/22/2016 |
| Proprietary Name | 1) Elecsys Cortisol II<br>2) Cortisol II CalSet |
| Common Name | 1) Cortisol II immunoassay<br>2) Cortisol II CalSet |
| Classification Name | 1) Enzyme Immunoassay, Cortisol<br>2) Secondary, calibrator |
| Product Codes | 1) JFT, 862.1205<br>2) JIT, 862.1150 |
| Predicate Devices | Elecsys Cortisol (k070788)<br>Elecsys Cortisol CalSet (k070788) |
| Establishment Registration | Roche Diagnostics GmbH in Mannheim, Germany, is 9610126<br>Roche Diagnostics GmbH in Penzberg, Germany, is 9610529<br>Roche Diagnostics in the United States is 1823260 |

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#### 1. DEVICE DESCRIPTION

The Elecsys Cortisol II assay makes use of a competition test principle using a monoclonal antibody which is specifically directed against cortisol. Endogenous cortisol which has been liberated from binding proteins with danazol competes with exogenous cortisol derivative in the test which has been labeled with ruthenium complex for the binding sites on the biotinylated antibody.

Results are determined via a calibration curve which is instrument specifically generated by 2point calibration and a master curve provided via reagent barcode.

#### 1.1. Reagents

The reagent working solutions include:

- rackpack (kit placed on instrument) .
- Streptavidin coated microparticles, ş
- Reagent 1 (Anti-cortisol-Ab~biotin) and ş
- Reagent 2 (Cortisol-peptide~Ru(bpy)2+3). ş

#### Calibrator 1.2.

The Cortisol II CalSet is a lyophilized human serum with added cortisol in two concentration ranges.

The CalSet includes:

- Cal 1 (approximately 12.5 nmol/L cortisol in a human serum matrix) .
- Cal 2 (approximately 1000 nmol/L cortisol in a human serum matrix) .

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#### 2. INDICATIONS FOR USE

#### 2.1. Immunoassay

Immunoassay for the in vitro quantitative determination of cortisol in human serum, and plasma. The determination of cortisol is used for the recognition and treatment of functional disorders of the adrenal gland.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys and cobas e immunoassay analyzers.

#### 2.2. Calibrator

Cortisol II CalSet is used for calibrating the quantitative Elecsys Cortisol II assay on the Elecsys and cobas e immunoassay analyzers.

#### TECHNOLOGICAL CHARACTERISTICS 3.

| Table 1: | | Assay Comparison |
|----------|--|------------------|
|----------|--|------------------|

| Assay Comparison | | |
|------------------------|-------------------------------------------------|---------------------------------------|
| Feature | Predicate Device: Elecsys Cortisol<br>(k070788) | Candidate Device: Elecsys Cortisol II |
| General Assay Features | | |

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| Assay Comparison | | |
|-----------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use/<br>Indications<br>for Use | Immunoassay for the in vitro quantitative<br>determination of cortisol in human serum,<br>plasma, urine and saliva. The determination<br>of cortisol is used for the recognition and<br>treatment of functional disorders of the<br>adrenal gland.<br>The electrochemiluminescence<br>immunoassay "ECLIA" is intended for use on<br>Elecsys and cobas e immunoassay<br>analyzers. | Immunoassay for the in vitro quantitative determination of cortisol in<br>human serum, and plasma. The determination of cortisol is used for<br>the recognition and treatment of functional disorders of the adrenal<br>gland.<br>The electrochemiluminescence immunoassay "ECLIA" is intended<br>for use on Elecsys and cobas e immunoassay analyzers. |
| Assay<br>Protocol | The Elecsys Cortisol assay makes use of a<br>competition test principle using a polyclonal<br>antibody which is specifically directed<br>against cortisol. Endogenous cortisol which<br>has been liberated from binding proteins with<br>danazol competes with exogenous cortisol<br>derivative in the test which has been labeled<br>with ruthenium complex for the binding sites<br>on the biotinylated antibody. | The Elecsys Cortisol II assay makes use of a competition test<br>principle using a monoclonal antibody which is specifically directed<br>against cortisol. Endogenous cortisol which has been liberated from<br>binding proteins with danazol competes with exogenous cortisol<br>derivative in the test which has been labeled with ruthenium<br>complex for the binding sites on the biotinylated antibody. |
| Detection<br>Protocol | Electrochemiluminescent Assay | Electrochemiluminescent Assay. |
| Applications | 18 minute application | 18 minute application. |
| Instrument<br>Platform | Elecsys 1010, Elecsys 2010, MODULAR<br>ANALYTIC E170 and cobas e analyzers. | cobas e 411 |
| Sample<br>Volume | 20 µL | 10 µL. |
| Sample<br>Type | Human serum, plasma, saliva and urine. | Human serum, and plasma. |

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| | Assay Comparison | |
|---------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Reagents | M Streptavidin-coated microparticles:<br>Streptavidin-coated microparticles;<br>preservative.<br>R1 Anti-cortisol-Ab~biotin: Biotinylated<br>polyclonal anti-cortisol antibody (ovine);<br>danazol; MES buffer; preservative.<br>R2 Cortisol-peptide~Ru(bpy): Cortisol<br>derivative (synthetic), labeled with ruthenium<br>complex; danazol; MES buffer; preservative. | M Streptavidin-coated microparticles: Streptavidin-coated<br>microparticles; preservative.<br>R1 Anti-cortisol-Ab~biotin: Biotinylated monoclonal anti-cortisol<br>antibody (ovine); danazol; MES buffer; preservative.<br>R2 Cortisol-peptide~Ru(bpy): Cortisol derivative (synthetic),<br>labeled with ruthenium complex; danazol; MES buffer;<br>preservative. |
| Calibrator | Elecsys Cortisol CalSet | Elecsys Cortisol II CalSet |
| Calibration<br>Interval | Calibration must be performed once per<br>reagent lot using fresh reagent (i.e. not more<br>than 24 hours since the reagent kit was<br>registered on the analyzer). Renewed<br>calibration is recommended as follows:<br>After 28 days when using the same<br>reagent lot.<br>After 7 days (when using the same<br>reagent kit on the analyzer).<br>As required: e.g. quality control findings<br>outside the defined limits | Calibration must be performed once per reagent lot using fresh<br>reagent (i.e. not more than 24 hours since the reagent kit was<br>registered on the analyzer). Renewed calibration is recommended<br>as follows:<br>After 8 weeks when using the same reagent lot.<br>After 7 days (when using the same reagent kit on the analyzer).<br>As required: e.g. quality control findings outside the defined<br>limits |
| Controls | Elecsys PreciControl Universal | Elecsys PreciControl Universal |
| Traceability /<br>Standardizati<br>on | standardized against the Enzymun-Test<br>Cortisol method. | Standardized against the IRMM (Institute for Reference Materials<br>and Measurements)/IFCC 451 panel (ID GC/MS, isotope dilution-<br>gas chromatography/mass spectrometry). |
| Reagent<br>Stability | When stored and handled as directed,<br>reagents are stable until the expiration date.<br>After Opening at 2-8°C - 30 days<br>On the Analyzers - 30 days | Unopened: 2-8°C - Up to the stated expiration date.<br>After Opening at 2-8°C - 12 weeks<br>On the Analyzers – 8 weeks |
| Measuring<br>Range | 0.5 – 1750 nmol/L | 3.0- 1750 nmol/L |

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| Assay Comparison | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
|---------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|
| Precision | Elecsys 2010/cobas e 411:<br>Within-run | cobas e 411:<br>Serum<br>Within-run (will be labeled Repeatability) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| | Sample Mean (nmol/L) SD %CV HS 1 208 2.76 1.3 HS 2 561 7.40 1.3 HS 3 1268 14.0 1.1 PCU* 1 363 5.08 1.4 PCU* 2 865 8.54 1.0 | | | | | | | | | | | | | | | | | | | | | | | | | Sample Mean (nmol/L) SD CV HS 1 3.09 0.219 7.1% HS 2 35.8 0.718 2.0% HS 3 283 7.29 2.6% HS 4 548 10.4 1.9% HS 5 1592 29.3 1.8% PCU* 1 308 4.33 1.4% PCU* 2 719 10.4 1.4% | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| | Total Sample Mean (µg/dL) SD %CV HS 1 208 3.29 1.6 HS 2 561 8.36 1.5 HS 3 1268 19.9 1.6 PCU* 1 363 5.67 1.6 PCU* 2 865 12.5 1.4 *PreciControl Universal | | | | | | | | | | | | | | | | | | | | | | | | | Total (will be labeled Intermediate precision) Sample Mean (nmol/L) SD CV HS 1 3.09 0.392 12.7% HS 2 35.8 1.36 3.8% HS 3 283 9.39 3.3% HS 4 548 17.4 3.2% HS 5 1592 42.7 2.7% PCU* 1 308 8.35 2.7% PCU* 2 719 18.0 2.5% *PreciControl Universal | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| Analytical<br>Sensitivity | Limit of Detection = <0.500 nmol/L | Limit of Blank (LoB): = 1.0 nmol/mL<br>Limit of Detection (LoD): = 1.5 nmol/mL<br>Limit of Quantitation (LoQ): = 3.0 nmol/mL | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |

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| Analytical<br>Specificity | Compound | Concentration<br>(µg/mL) | %<br>Cross-<br>Reactivity | Cross reactant | concentration<br>tested<br>(µg/mL) | Highest<br>cross-<br>reactivity<br>observed<br>(%) |
|---------------------------|-----------------------------|--------------------------|---------------------------|------------------------------|------------------------------------|----------------------------------------------------|
| | Allotetrahydro-<br>Cortisol | 0.1 | 165 | 11-<br>Deoxycorticosterone | 10 | 0.640 |
| | Corticosterone | 10 | 5.8 | 11-<br>Deoxycortisol | 10 | 4.90 |
| | Cortisol 21<br>-sulfate | 10 | 0.04 | 17-α-<br>Hydroxyprogesterone | 10 | 0.080 |
| | Cortisone | 10 | 0.30 | Corticosterone | 10 | 2.48 |
| | 11-Deoxycortl-<br>costerone | 10 | 0.69 | Cortisone | 10 | 6.58 |
| | 11-Deoxycortisol | 10 | 4.1 | Dexamethasone | 10 | Not<br>detectable |
| | Dexamethasone | 10 | 0.08 | Fludrocortisone | 10 | 0.200 |
| | 21-deoxycortisol | 1 | 45.4 | Prednisone | 10 | 2.23 |
| | 6-b-Hydroxycortisol | 1 | 158 | Progesterone | 10 | 0.035 |
| | 17-Hydroxy-<br>progesterone | 10 | 1.5 | 21-<br>Deoxycortisol | 1 | 2.40 |
| | 6-a-methyl-<br>prednisolone | 0.1 | 389 | Prednisolone | 0.1 | 7.98 |
| | Prednisolone | 0.1 | 171 | 6-α-<br>Methylprednisolone | 0.1 | 12.0 |
| | Prednisone | 10 | 0.28 | Androstendion | 10 | 0.1 |
| | Progesterone | 10 | 0.35 | Estradiol | 10 | 0.2 |
| | | | | Estriol | 10 | Not<br>detectable |
| | | | | Estron | 10 | 0.8 |
| Linearity | 1 to 59.8 µg/dL | | | 3.0 to 1750 nmol/mL | | |

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| Assay Comparison | | |
|------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Limitations | When performed in serum and plasma, the assay is unaffected by:<br>Bilirubin <60 mg/dL<br>Hemolysis < 1.9 g/dL<br>Lipemia < 2700 mg/dL<br>Biotin< 30 ng/mL<br>Rheumatoid factors < 1100 IU/mL<br>In vitro tests were performed on 17<br>commonly used pharmaceuticals. No<br>interference with the assay was found.<br>The risk of interference from potential<br>immunological interactions between test<br>components and rare sera has been<br>minimized by the inclusion of suitable<br>additives. In rare cases, interference due<br>to extremely high titers of antibodies to<br>analyte-specific antibodies, streptavidin<br>or ruthenium can occur. The test<br>contains additives which minimize these<br>effects.<br>Pregnancy, contraceptives and estrogen<br>therapy give rise to elevated cortisol<br>concentrations.<br>In samples from patients who have been<br>treated with prednisolone, 6 α<br>Methylprednisolone or prednisone, falsely<br>elevated concentrations of cortisol may<br>be determined.<br>During metyrapon tests, 11 deoxycortisol<br>levels are elevated. Falsely elevated<br>cortisol values may be deter-mined due<br>to cross reactions (see section on<br>analytical specificity).<br>Patients suffering from 21 hydroxylase<br>deficiency exhibit elevated 21<br>deoxycortisol levels and this can also<br>give rise to falsely elevated cortisol<br>results.<br>The time of sample collection must be<br>taken into account when interpreting<br>results due to the cortisol secre-tion<br>circadian rhythm. Severe stress can also<br>give rise to elevated cortisol levels.<br>For diagnostic purposes, the results should<br>always be assessed in conjunction with the<br>patient's medical history, clinical examination<br>and other findings. | When performed in serum and plasma, the assay is unaffected by:<br>Bilirubin ≤ 25 mg/dL<br>Hemolysis ≤ 0.5 g/dL<br>Lipemia ≤ 1500 mg/dL<br>Biotin ≤ 30 ng/mL<br>Rheumatoid factors < 600 IU/mL<br>IgG ≤ 50 g/L<br>IgM ≤ 10 g/L<br>IgA ≤ 10 g/L<br>In vitro tests were performed on 16 commonly used<br>pharmaceuticals. No interference with the assay was found.<br>In rare cases, interference due to extremely high titers of<br>antibodies to analyte-specific antibodies, streptavidin or<br>ruthenium can occur. These effects are minimized by suitable<br>test design.<br>Pregnancy, contraceptives and estrogen therapy give rise to<br>elevated cortisol concentrations.<br>In samples from patients who have been treated with<br>prednisolone, 6 α Methylprednisolone or prednisone, falsely<br>elevated concentrations of cortisol may be determined.<br>During metyrapon tests, 11 deoxycortisol levels are elevated.<br>Falsely elevated cortisol values may be deter-mined due to<br>cross reactions (see section on analytical specificity).<br>Patients suffering from 21 hydroxylase deficiency exhibit<br>elevated 21 deoxycortisol levels and this can also give rise to<br>falsely elevated cortisol results.<br>The time of sample collection must be taken into account when<br>interpreting results due to the cortisol secre-tion circadian<br>rhythm. Severe stress can also give rise to elevated cortisol<br>levels.<br>For diagnostic purposes, the results should always be assessed in<br>conjunction with the patient's medical history, clinical examination<br>and other findings. |

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### Assay Comparison

| Labeled Performance Characteristics | | | | | | | | | | |
|---------------------------------------------------------------------|---------------------------------------------------|----------------|--------|--------------------------------------------|------|--------|------------------------------------------------|---------------------|----------------------|-----------------------|
| Method<br>Comparison<br>(LC/MS) | n = 208<br>Min = 10.9 nmol/L<br>Max = 1599 nmol/L | | | Passing/Bablok<br>1.022<br>2.92<br>0.930 | | | Deming regression<br>1.055<br>-6.10<br>0.993 | | | |
| Method<br>Comparison<br>(Cortisol II (y)<br>versus Cortisol<br>(x)) | n = 536<br>Min = 1.54 nmol/L<br>Max = 1680 nmol/L | | | Passing/Bablok<br>0.758<br>10.20<br>0.8725 | | | Deming regression<br>0.806<br>-10.31<br>0.9676 | | | |
| | Subject<br>cohort | Sample<br>size | Min | Max | Mean | Median | 2.5th percen-<br>tile | 5th percen-<br>tile | 95th percen-<br>tile | 97.5th<br>percen-tile |
| Reference<br>range<br>serum/plasma | | N | nmol/L | | | | | | | |
| | 6 - 10 am | 296 | 23.7 | 835 | 316 | 303 | 133 | 166 | 507 | 537 |
| | LCL - UCL* | - | - | - | - | - | 70.1 - 161 | 133 - 183 | 488 - 537 | 511 - 584 |
| | 4 - 8 pm | 300 | 19.1 | 544 | 172 | 161 | 68.2 | 73.8 | 291 | 327 |
| | LCL - UCL* | - | - | - | - | - | 46 - 72 | 68 - 87 | 269 - 327 | 301 - 401 |

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| Characteristic | Predicate device: Elecsys Cortisol CalSet<br>(k070788) | Candidate device: Cortisol II CalSet |
|----------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | Cortisol CalSet is used for calibrating the<br>quantitative Elecsys Cortisol assay on the<br>Elecsys and cobas e immunoassay<br>analyzers. | Cortisol II CalSet is used for calibrating the<br>quantitative Elecsys Cortisol II<br>assay on the Elecsys and cobas e<br>immunoassay analyzers. |
| Analyte | Cortisol (synthetic) | Same |
| Matrix | Human serum matrix | Same |
| Levels | Two | Same |
| Target Ranges | Cal 1: 12.5 nmol/L<br>Cal 2: 1000 nmol/L | Same. |
| Format | Lyophilized | Same |
| Traceability | standardized against the Enzymun-Test<br>Cortisol method. | The Elecsys Cortisol II assay has been<br>standardized against the IRMM(Institute for<br>Reference Materials and<br>Measurements)/IFCC-451 panel (ID-GC/MS,<br>isotope dilution- gas chromatography/mass<br>spectrometry) |
| Handling | Carefully dissolve the contents of one bottle<br>by adding exactly 1.0 mL of distilled or<br>deionized water and allow to stand closed for<br>15 minutes to reconstitute. Mix carefully,<br>avoiding foam formation.<br>Transfer aliquots of the reconstituted<br>calibrators into empty labeled snap-cap<br>bottles (CalSet Vials). Attach the supplied<br>labels to the additional bottles. Store the<br>aliquots immediately at -20 °C.<br>Perform only one calibration procedure per<br>aliquot. | Same. |

### Table 2: CalSet Comparison

#### 4. NON-CLINICAL PERFORMANCE EVALUATION

Non-clinical performance evaluations for the Elecsys Cortisol II executed with the study briefly summarized.

#### 4.1. Precision (Human Serum)

Precision of the Elecsys Cortisol II assay was evaluated on one cobas e 411 Immunoassay Analyzer according to CLSI EP5-A2 guideline. One reagent lot was evaluated.

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The protocol consisted of testing 2 replicates each of the controls (PC Universal) and 5 human serum samples (HS) per run, 2 runs per day for 21 days. The samples were run in randomized order on the analyzer. Samples used were native, diluted and spiked human serum pools.

Repeatability and Intermediate precision were calculated according to EP5-A2.

#### Limit of Blank 4.2.

For the analytical sensitivity studies, 2 cobas e 411 analyzers and 2 lots of reagents were used. The Limit of Blank (LoB) was determined using native human serum pools and Universal Diluent. A total of n = 60 LoB measurements were made (5 blank samples, 1 replicate, 2 runs per day on 2 instruments over 3 days). The LoB was calculated according to CLSI EP17-A2 (non-parametric approach).

#### Limit of Detection 4.3.

For the analytical sensitivity studies, 2 cobas e 411 analyzer and 2 lots of reagents were used. The Limit of Detection (LoD) was determined using 5 low-level human serum samples (diluted). A total of n = 60 LoD measurements were made (5 samples, 2 runs per day on 2 instruments over 3 days). The LoD was calculated according to CLSI EP17-A2.

#### Limit of Quantitation 4.4.

The Limit of Quantitation (LoQ) was determined using a set of nine human serum samples, three reagent lots on two cobas e 411 analyzer. The Limit of Quantitation (LoQ) was determined in accordance with CLSI Guideline EP17-A2. Each sample was analyzed in replicates of 2, two runs per day over 3 days.

#### Linearity 4.5.

### 4.5.1. Serum

The linearity results were obtained with serum samples on the cobas e 411 Immunoassay analyzer. A high analyte serum sample (pooled human serum sample, spiked) was diluted with Diluent Universal (protein matrix). 19 concentrations (thereof 17 dilutions) throughout the

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measuring range were prepared. Samples were assayed in 3-fold determination within a single run.

The linearity data were analyzed with regards to linear, quadratic and cubic polynomials according to CLSI EP6-A. In a first step, a linearity check was performed with a first order (linear) regression and then with higher order models (quadratic and cubic).

# 4.5.2.

Linearity of the Elecsys Cortisol II assay was assessed on the cobas e 411 Immunoassay Analyzer according to CLSI EP6-A.

A high analyte plasma sample (pooled human plasma sample, spiked) was diluted with Diluent Universal (protein matrix). 19 concentrations (thereof 17 dilutions) throughout the measuring range were prepared. Samples were assayed in 3-fold determination within a single run.

The linearity data were analyzed with regards to linear, quadratic and cubic polynomials according to CLSI EP6-A. In a first step, a linearity check was performed with a first order (linear) regression and then with higher order models (quadratic and cubic).

#### Analytical Specificity 4.6.

### 4.6.1. Serum/Plasma

The specificity of the Elecsys Cortisol II assay was determined using two human serum sample pools (diluted as well as spiked) spiked with potential cross-reactant compounds. The analyte concentration of the samples was at approx. 60 and 300 µg/L Cortisol (conversion factor: µg/L x 2.7586 = nmol/L). The spiked and non-spiked samples were tested in duplicates on the cobas e 411 Immunoassay Analyzer. Cross-reactivity results will be reported in the method sheet.

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#### Endogenous Interferences 4.7.

The effect on quantitation of analyte in the presence of endogenous interfering substances using the Elecsys Cortisol II assay was determined on the cobas e 411 Immunoassay analyzer using human serum samples (single samples as well as pools, native, diluted and spiked).

For each interfering substance three human serum samples containing low, mid and high concentrations of Cortisol were tested. The recovery for each sample was calculated by comparison to the reference sample. Predefined acceptance criterion was met. The claims included in the method sheet were set to the specification concentration.

#### 4.8. HASA Effect

The effect of the presence of human anti-sheep antibodies (HASA) on the Elecsys Cortisol II assay was assessed on the cobas e 411 analyzer.

As no native human sample with this type of interference was available the effect of human antisheep antibodies was investigated in a model system using donkey anti-sheep polyclonal antibodies (DASA).

Two serum samples with cortisol concentrations of 173 respectively 789 nmol/L were divided into two aliquots each. One aliquot was spiked with the 20ug/mL DASA and was then diluted with the unspiked aliqout of the serum sample in 10% increments.

The recovery for each sample was calculated by comparison to the unspiked reference sample.

#### Exogenous Interferences- Drugs 4.9.

The effect on quantitation of analyte in the presence of drugs was determined by comparing values obtained from samples spiked with 16 pharmaceutical compounds spiked into two human serum samples (native, diluted and spiked human serum pools as well as single samples) and tested on the cobas e 411 Immunoassay Analyzer. The analyte concentrations of the samples were approximately 170 and 750 nmol/L. The drug concentrations tested are in accordance with the recommendation (if available) given in the CLSI guideline EP7-A2. The two serum samples were divided into aliquots and spiked with the potential interferents. The reference sample without interferent was spiked with the respective amount of solvent only.

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The Cortisol II concentration of the spiked aliquots was determined in 3-fold determination and compared to the Cortisol concentration determined for the reference aliquot (also in 3-fold determination) on one cobas e 411 Immunoassay Analyzer.

# 4.10. Exogenous Interferences- Anticoagulants

The effect on quantitation of analyte in the presence of anticoagulants with the Elecsys Cortisol II Immunoassay was determined by comparing values obtained from samples (native, diluted and spiked human serum pools as well as single samples) drawn into Serum, Li-Heparin, K2-EDTA-, K3-EDTA-plasma primary tubes and Li-Heparin Plasma Gel Separation Tubes. A minimum of 48 serum/plasma pairs per sample material were tested in duplicate with one reagent lot on one cobas e 411 Immunoassay Analyzer. Potential effects are assessed by regression analysis.

# 4.11. Method Comparison

To show correlation, the following two method comparisons were performed:

Method Comparison 1: Elecsys Cortisol II vs. LC-MS (Reference Method)

Method Comparison 2: Elecsys Cortisol II vs. Cortisol I (k070788)

4.11.1. Method Comparison 1: Elecsys Cortisol II vs. LC-MS (Reference Method)

A total of 208 human serum samples (all native single donors) were measured in singleton covering the entire measuring range. The study was performed on the cobas e 411 (Y) and the LC-MS method (X) at UZ Gent, Lab voor Klinische Biologie (Gent, Belgium).

4.11.2. Method Comparison 2: Elecsys Cortisol II vs. Cortisol I (k070788)

The second method comparison shows the correlation of the Elecsys Cortisol II to the current Elecsys Cortisol assay. The expected differences due the improved specificity of the Elecsys Cortisol II assay are shown. The study was performed on the cobas e 411 (Y and X) analyzer at UZ Gent, LMU Großhadern, Uni Klinik Leipzig and Labor Limbach, respectively. A total of 536 human serum samples (all native single donors) were measured.

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# 4.12. Reagent Stability

To test reagent stability, three studies were executed with two studies completed.

### 4.12.1. Study 1. Reagent stability onboard (8 weeks)

Reagent onboard stability for the Elecsys Cortisol II assay was tested on one cobas e 411 Immunoassay Analyzer.

A fresh reagent rackpack was placed on the analyzer and calibrated. Reference values for the samples tested were determined. After measurement the rackpack was removed from the analyzer and kept at 20°C + 3°C (on-board condition). On day 8, 36 and day 64, the same samples were measured with this reagent kit using the calibration curves established on day 1, 29 and 57, respectively.

Samples tested in duplicate include five human serum (HS) samples and two controls. Native, diluted and spiked human serum samples were used.

# 4.12.2. Study 2. Reagent stability after first opening at 2-8°C (84 days)

Reagent stability after first opening for the Elecsys Cortisol II assay was tested on one cobas e 411 Immunoassay Analyzer.

A fresh reagent rackpack was placed on the analyzer and calibrated. Reference values for the samples tested were determined. After measurement the kit was removed from the analyzer and kept at 2-8 °C for 29 days, 57 days and 92 days. After 29 days, 57 days and 92 days the kit was placed on the analyzer again, calibrated and the test samples were determined.

Samples tested in duplicate include five human serum (HS) samples and two controls. Native, diluted and spiked human serum samples were used.

4.12.3. Study 3: A real-time stability study is ongoing to support shelf-life stability claim. In the ongoing real-time stability study, the Elecsys Cortisol II assay material is stored at 2-8°C. The stored assay reagents are tested at time point T=0 and at specified intervals over the shelf life of the device up to the planned shelf life plus one month. Testing will be performed using

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PreciControl Universal. Level 1 and 2 (lyophylized, stored at -20°C) and human serum samples (stored at -80°C).

Data for the time-points at 0, 13, 16 and 19 months for MP lot and at 0, 7, 16 and 19 months for P2 and P3 lot tested in duplicate will be available. The average on-test recovery value will be calculated as percent recovery compared to the reference value (Assigned value for PreciControl Universal, Level 1 and 2).

# 4.13. Sample Stability

To test sample stability, three studies were completed. Because these studies are not analyzer dependent, and the assay is a global product currently available rest of world (ROW), the studies were executed on the cobas e 601. Study results can be applied to the cobas e 411 since sample stability is independent of the analyzer. For each study, the samples used were all single donors (native, spiked and diluted) and the following acceptance criteria was applied.

### 4.13.1. Study 1. Sample stability at 2-8°C

Twelve human serum, Li-Heparin, K2-EDTA, and K3-EDTA plasma samples were aliquoted and measured fresh (reference value) and after storage at 2-8°C for 4 days.

Measurements were performed with three-fold determination on a cobas e 601 Immunoassay Analyzer and recovery was calculated as percent of the reference value.

The human serum samples used were native, spiked and diluted single donors.

Study 2. Sample stability at Room Temperature (15-25°C) 4.13.2.

Twelve human serum, Li-Heparin, K2-EDTA, and K3-EDTA plasma samples were aliquoted and measured fresh (reference value) and after storage at 20-25°C for 24 hours.

Measurements were performed with three-fold determination on a cobas e 601 Immunoassay

Analyzer and recovery was calculated as percent of the reference value.

The human serum samples used were native, spiked and diluted single donors.

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### 4.13.3. Study 3. Sample stability -15 to -25°C

Twelve human serum. Li-Heparin, K2-EDTA, and K3-EDTA_plasma samples were aliquoted and measured fresh (reference value) and after storage of 12 months at -15 to -25°C.

Measurements were performed with three-fold determination on a cobas e 601 Immunoassay Analyzer and recovery was calculated as percent of the reference value.

The human serum samples used were native, spiked and diluted single donors.

# 4.14. Calibration Stability

To test calibration stability, two studies were completed, including:

- . Study 1. Lot calibration stability
- Study 2. On-board calibration stability

# 4.14.1. Study 1

Calibration of an Elecsys Cortisol II reagent lot is recommended every 8 weeks. During that time period fresh reagent kits of the same lot can be used without calibration using the calibration curve of the Day 1 reagent kit.

Elecsys Cortisol II was calibrated with a fresh reagent kit on Day 1 using a cobas e 411 Immunoassay Analyzer. After 29 days, 57 days and 92 days a new reagent kit of the same lot was used and recovery of samples was determined using the calibration curve of day 1.

Five human serum (HS) samples and two control samples were tested; each sample was tested with two-fold determination. The human serum samples used were native, diluted and spiked.

# 4.14.2. Study 2

Reagent Calibration Stability for the Elecsys Cortisol II assay was tested on one cobas e 411 Immunoassay Analyzer.

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A fresh Reagent Rack-Pack was placed on the analyzer and calibrated. All samples were measured on day 1. On day 8 the same samples were measured with the same reagent kit kept at 20°C + 3°C (on-board condition) using the calibration curves established on day 1.

Five human serum (HS) samples and two control samples were tested; each sample was tested with two-fold determination. The human serum samples used were native, diluted and spiked.

# 4.15. Calibrator studies

The Elecsys Cortisol II CalSet was evaluated for value assignment, reconstitution and stability.

### 4.15.1. Value assignment

Value assignment testing was conducted and passed pre-defined acceptance criteria. The target values for the two levels of the Cortisol II CalSet kit are chosen to obtain the best fit with the Master Calibration Curve, together with the Rodbard curve parameters encoded in the reagent barcode. For each Elecsys Cortisol II CalSet lot manufactured, the calibrators are run in duplicate on at least three (3) cobas e 411 analyzers and at least three (3) cobas e 601/cobas e 602/MODULAR ANALYTICS E170 analyzers with all Cortisol II reagent lots available. The assigned value of each calibrator is defined as the mean value obtained over at least six (6) runs on at least three (3) analyzers) of the respective calibrator.

Measurement values for PreciControl Universal (Levels 1 & 2), a multi-analyte control recommended for use to monitor accuracy and precision of specified analytes, are read from the calibration curves generated. The pre-defined acceptance criteria for PreciControl Universal have to be met to release the Assigned Values for Cortisol II CalSet.

### 4.15.2. Reconstitution

Reconstitution time for the lyophilized Cortisol II CalSet was tested. Two sets of Cortisol II CalSet were reconstituted, one for 15 minutes and the other for 30 minutes. Signal recovery after 30 minutes reconstitution was compared to the signal value after 15 minutes.

Cortisol II CalSet was evaluated in duplicate on the cobas e 411 analyzer.

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### 4.15.3. Stability

Three studies were performed in order to verify the stability claims for the Cortisol II CalSet. Stability studies after reconstitution and an accelerated stability study were completed on the Elecsys 2010.

### 4.15.3.1. Study 1- Stability at -20°C (after reconstitution)

The on-test and reference materials were tested in duplicate on the Elecsys 2010. The on-test material was reconstituted and stored in closed vials for 3 months at -20°C. The on-test signal recovery was calculated as percent of the reference value.

### 4.15.3.2. Study 2- Onboard stability at 20-25°C after reconstitution

The on-test and reference materials were tested in duplicate on the Elecsys 2010. The on-test material was reconstituted and stored in open vials for 4 hours at 20-25°C. The on-test signal recovery was calculated as percent of the reference value.

### 4.15.3.3. Real-time Stability

Since there was no change in formulation and target values to the Cortisol II CalSet the real-time stability data (18 months at 2-8°C) of the current Cortisol CalSet is applicable.

In the real-time stability study, the Cortisol CalSet on-test material was stored at 2 - 8°C. The CalSets were tested at specified intervals over the shelf life of the device up to the planned shelf life plus one month.

Data for the time-points after manufacturing, 7 (Lot 1), 6 (Lot 2 & 3), 14 (Lot 1), 13 (Lot 2 & 3), and 19 months were tested in duplicate. The stability of the CalSet was assessed via accurate recovery of PreciControl Universal.

### 4.15.3.4. Reconstitution study

Reconstitution time for the lyophilized Cortisol II CalSet was tested. Two sets of Cortisol II CalSet were reconstituted, one for 15 minutes and the other for 30 minutes. Signal recovery after

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30 minutes reconstitution was compared to the signal value after 15 minutes. Cortisol II CalSet was evaluated in duplicate on the cobas e 411 analyzer.

#### CLINICAL PERFORMANCE EVALUATION 5.

Clinical samples were collected at three sites in the United States in order to establish the reference range values for the Elecsys Cortisol II assay. Reference ranges for self-reported healthy males and females were determined using the median value, the 5th -95th and the 2.5th -97.5th percentiles (nmol/L) as lower and upper limit of normal, respectively. The evaluation was done at one site in St Louis, Missouri with one reagent lot (MP) using one cobas e 411 analyzer. Samples were all native human serum samples measured in singleton, over 2 runs within 10 days.

#### 6. ADDITIONAL INFORMATION

Two additional products are used with the Elecsys Cortisol II assay; Cortisol CalCheck and the PreciControl Universal. There has been no change to the manufacturing process, value assignment process, product stability or specifications of these products.

The Elecsys Cortisol CalCheck labeling has been updated to incorporate the Cortisol II information and has been included for review.

#### 7. CONCLUSIONS

The information provided in this Premarket Notification [510(k)] will support a determination of substantial equivalence for the Elecsys Cortisol II, and Elecsys Cortisol II CalSet. The data supports a safe, effective device which performs as well as or better than the predicate devices.