CORTISOL ELISA

K062626 · Ibl-Hamburg GmbH · CGR · Dec 20, 2006 · Clinical Chemistry

Device Facts

• Record ID: K062626
• Device Name: CORTISOL ELISA
• Applicant: Ibl-Hamburg GmbH
• Product Code: CGR · Clinical Chemistry
• Decision Date: Dec 20, 2006
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1205
• Device Class: Class 2

Intended Use

The IBL Cortisol enzyme linked immunosorbent assay is for the in-vitro-diagnostic quantitative determination of cortisol in human serum and saliva. The Cortisol ELISA kit is useful as an aid in the differential diagnosis of Cushing syndrome and Addison’s disease.

Device Story

The IBL Cortisol ELISA is an in-vitro diagnostic test for quantitative determination of cortisol in human saliva and diluted serum. It utilizes a solid-phase enzyme-linked immunosorbent assay (ELISA) based on a competitive binding principle. Patient samples and enzyme-labeled cortisol compete for binding sites on antibody-coated microtiter wells. After incubation and washing, a substrate reaction occurs; the resulting color intensity is inversely proportional to the cortisol concentration in the sample. Results are determined via a standard curve. The device is intended for use by clinical laboratory professionals to aid in the assessment of Cushing Syndrome and Addison's Disease. Healthcare providers use the quantitative results to support clinical diagnosis. The device provides a standardized method for measuring cortisol levels, facilitating clinical decision-making regarding adrenal function.

Clinical Evidence

No clinical studies performed. Evidence consists of analytical performance data: intra-assay precision (mean CV 5.7% saliva, 8.8% serum), inter-assay precision (mean CV 7.4% saliva, 11.2% serum), and linearity (mean recovery 97% saliva, 105% serum). Method comparison against predicate using 129 serum and 130 saliva samples showed high correlation (R² ≥ 0.990).

Technological Characteristics

Solid-phase competitive ELISA. Components: antibody-coated microtiter strips, enzyme conjugate, standards, controls, wash buffer, TMB substrate, and stop solution. Connectivity: standalone. Energy source: none (manual/automated plate reader). Sterilization: N/A. Software: N/A (manual calculation/plate reader processing).

Indications for Use

Indicated for the in-vitro diagnostic quantitative determination of cortisol in human serum and saliva as an aid in the differential diagnosis of Cushing syndrome and Addison's disease.

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

• IBL Cortisol LIA TEST KIT (k052359)

Submission Summary (Full Text)

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K062626

## SUMMARY OF SAFETY AND EFFECTIVENESS FOR IBL CORTISOL ELISA

DEC 2 0 2006

| Manufacturer: | IBL. Immuno Biological Laboratories<br>Flughafenstrasse 52A, D-22335<br>Hamburg, Germany |
|----------------------|----------------------------------------------------------------------------------------------------------|
| Contact Information: | Victor Herbst<br>IBL Immuno Biological Laboratories<br>Flughafenstrasse 52A, D-22335<br>Hamburg, Germany |

#### Device Name / Classification:

The device trade name is the IBL Cortisol ELISA having FDA assigned name: Cortisol (hydrocortisone and hydroxycorticosterone) test system, 21 CFR, 862.1205, categorized as Class II medical devices for the Clinical Chemistry Panel, as Product Code CGR.

### Test Principle

Solid phase enzyme-linked immunosorbent assay (ELISA) based on the competition principle. An unknown amount of antigen present in the sample and a fixed amount of enzyme labelled antigen compete for the binding sites of the antibodies coated onto the wells. After incubation the wells are washed to stop the competition reaction. After the substrate reaction the intensity of the developed color is inversely proportional to the amount of the antigen in the sample. Results of samples can be determined directly using the standard curve.

#### Device Intended Use:

Solid phase enzyme-linked immunosorbent assay for the in-vitro diagnostic quantitative determination of free Cortisol in human saliva and of total Cortisol in diluted serum as an aid in the assessment of Cushing Syndrome and Addison's Disease.

#### Device Performance

All technical data are included in this 510(k) submission. The normal ranges and stability data will be overtaken from the original Cortisol LIA submission No. K052359 (respectively No. K010790) which is manufactured in same way. All single components keep the same, except the substrate system which uses an ordinary TMB (Tetramethylbenzidine) substrate with a given shelf life of 18months by manufacturer. The shelf lifes are therefore as follows:

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## Stability of kit components at (2 - 8 ℃) :

| Microtiter strips | 12 months |
|----------------------------|-----------|
| Enzyme conjugate. | 9 months |
| Standard A-G | 9 months |
| Kit control 1, 2 | 9 months |
| Wash buffer. Concentrate | 12 months |
| ready to use TMB substrate | 18 months |
| TMB Stop solution | 36 months |

Therefore the complete Kit will have a shelf life of 9 months at 2 - 8 ℃.

## Method comparison

A comparison study was performed using 130 saliva and 1290 serum samples. These samples were tested on the IBL Cortisol ELISA and compared to the Cortiso! LIA, The results from measuring the samples in both methods yielded the following correlation:

| Method<br>Comparison<br>versus LIA | | Saliva | $IBL-ELISA = 0.92 x IBL-Luminescence IA + 0.06 µg/dL$ | r = 0.995;n = 130 |
|------------------------------------|--|--------|-------------------------------------------------------|--------------------|
| | | Serum | $IBL-ELISA = 1.17 x IBL-Luminescence IA - 2.2 µg/dL$ | r = 0.997; n = 129 |

Additionally 33 serum samples from the DGKC (Deutsche Gesllschaft für klinische Chemie, Bonn Germany) quality assessment scheme for hormones which were obtained using a GC/MS method, according to: Siekmann et al., J.Clin.Chem.Clin.Biochem. 20 (1982) 883-892, were used for comparison study to the given GC/MS reference values. The results from measuring the samples yielded the following correlation:

| Method<br>Comparison<br>versus GC/MS | Serum | $IBL-ELISA = 0.97 x GCMS + 2.3 \mu g/dL$ | r = 0.982; n = 33 |
|--------------------------------------|-------|------------------------------------------|-------------------|
|--------------------------------------|-------|------------------------------------------|-------------------|

#### Interference Studies

The following blood components have been tested in serum and saliva and do not have a significant effect (+/- 20 % of expected) on the test results up to the concentrations stated below:

| | Serum | |
|--------------|-----------|------------------|
| | Conc. | Cortisol (µg/dL) |
| Hemoglobin | 4.0 mg/mL | 0.06; 0.33; 0.62 |
| Bilirubin | 0.5 mg/mL | 0.07; 0.35; 0.63 |
| Triglyceride | 30 mg/mL | 0.07; 0.40; 0.75 |
| | Saliva | |
| | Conc. | Cortisol (µg/dL) |
| Thimerosal | 0.50 % | 0.19; 0.25; 0.34 |
| Blood | 0.125 % | 0.09; 0.26 |
| NaN3 | 0.60 % | 0.23; 0.31 |

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## The overall performance of the IBL Cortisol ELISA is:

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:

:

| | Substance | Cross<br>Reactivity (%) | | | | | |
|------------------------------------------------------|-------------------------------|-----------------------------------|------------------------------------------------------------------|------------------|-------------------------|-------------|-----|
| Analytical<br>Specificity<br>(Cross<br>Reactivity) | Prednisolone | 29 | Cross-reactivity<br>of other<br>substances<br>tested<br>< 0.01 % | | | | |
| | 11-Desoxy-Cortisol | 16 | | | | | |
| | Corticosterone | 2.4 | | | | | |
| | Cortisone | 3.3 | | | | | |
| | Prednisone | 2.2 | | | | | |
| | 17α-OH-Progesterone | 1.2 | | | | | |
| | Desoxy-Corticosterone | 0.5 | | | | | |
| | 6α-Methyl-17α-OH-Progesterone | 0.3 | | | | | |
| Analytical<br>Sensitivity<br>(Limit of<br>Detection) | 0.015 µg/dL | Mean signal (Zero-Standard) - 2SD | | | | | |
| Functional<br>Sensitivity | 0.060 µg/dL | Mean Conc. < 20 % CV | | | | | |
| | | Saliva (n = 20) | Serum (1:50 diluted; n = 20) | | | | |
| Precision | Conc.<br>(µg/dL) | SD<br>(µg/dL) | CV<br>(%) | Conc.<br>(µg/dL) | SD<br>(µg/dL) | CV<br>(%) | |
| Intra-Assay | 0.252 | 0.016 | 6.4 | 0.103 | 0.012 | 11.8 | |
| | 0.312 | 0.024 | 7.6 | 0.499 | 0.053 | 10.7 | |
| | 2.927 | 0.094 | 3.2 | 3.421 | 0.132 | 3.8 | |
| | 0.215 | 0.020 | 9.1 | 0.094 | 0.010 | 10.8 | |
| Inter-Assay | 0.864 | 0.059 | 6.9 | 0.394 | 0.043 | 10.9 | |
| | 2.638 | 0.164 | 6.2 | 0.582 | 0.070 | 12.0 | |
| | | Dilution | Saliva | | Dilution | Serum | |
| | | Meas.<br>(µg/dL) | Rec.<br>(%) | | Calc. (1:50)<br>(µg/dL) | Rec.<br>(%) | |
| Linearity | | 3.035 | 100 | | 1:50 | 35.2 | 100 |
| | | 1:2 | 1.259 | 83 | 1:100 | 16.8 | 96 |
| | | 1:4 | 0.635 | 84 | 1:200 | 9.4 | 107 |
| | | 1:8 | 0.340 | 90 | 1:400 | 4.4 | 101 |
| | | 1:16 | 0.184 | 97 | 1:800 | 2.4 | 111 |
| | | 1:32 | 0.108 | 114 | 1:1600 | 1.1 | 97 |
| | | - | 0.834 | 100 | 1:50 | 29.6 | 100 |
| | | 1:2 | 0.416 | 115 | 1:100 | 14.4 | 98 |
| | | 1:4 | 0.202 | 106 | 1:200 | 7.5 | 101 |
| | | 1:8 | 0.119 | 95 | 1:400 | 4.4 | 120 |
| | | - | 0.602 | 100 | 1:50 | 227.2 | 100 |
| | | 1:2 | 0.254 | 84 | 1:100 | 108.9 | 96 |
| | 1:4 | 0.146 | 97 | 1:200 | 51.4 | 90 | |
| | 1:8 | 0.082 | 109 | 1:400 | 25.8 | 91 | |
| | | | | 1:800 | 13.4 | 94 | |
| | | | | 1:1600 | 6.5 | 96 | |
| | | | | 1:3200 | 3.6 | 112 | |

:

.

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| | Saliva | | | | Serum | | | | | |
|----------------------|--------------------|---------|---------|-----------------------------------------------|---------|-------------------|---------|---------|--------------------|-----|
| Recovery | Conc. | Added | Meas. | Rec. | Conc. | Added | Meas. | Expect. | Rec. | |
| | (µg/dL) | (µg/dL) | (µg/dL) | (%) | (µg/dL) | (µg/dL) | (µg/dL) | (µg/dL) | (%) | |
| | Saliva 1<br>(0.25) | | 0.04 | 0.30 | 104 | Serum 1<br>(4.75) | 2.0 | 7.6 | 6.7 | 113 |
| | | | 0.08 | 0.34 | 105 | | 3.9 | 8.1 | 8.7 | 93 |
| | | | 0.16 | 0.39 | 97 | | 7.8 | 13.7 | 12.6 | 109 |
| | | | 0.31 | 0.61 | 109 | | 15.6 | 21.2 | 20.4 | 104 |
| | | | 0.63 | 0.91 | 104 | | 31.3 | 38.3 | 36.0 | 106 |
| | | | 1.25 | 1.51 | 101 | | 62.5 | 72.0 | 67.3 | 107 |
| | | | 2.50 | 2.19 | 80 | | 125.0 | 120.5 | 129.8 | 93 |
| | Saliva 2<br>(0.30) | | 0.03 | 0.30 | 91 | Serum 2<br>(23.0) | 2.0 | 22.1 | 25.0 | 89 |
| | | | 0.06 | 0.37 | 103 | | 3.9 | 23.6 | 26.9 | 88 |
| | | | 0.13 | 0.43 | 102 | | 7.8 | 27.6 | 30.8 | 89 |
| | | | 0.25 | 0.57 | 105 | | 15.6 | 37.6 | 38.6 | 97 |
| | | | 0.50 | 0.93 | 116 | | 31.3 | 56.6 | 54.3 | 104 |
| | | | 1.00 | 1.22 | 94 | | 62.5 | 83.1 | 85.5 | 97 |
| | | | 2.00 | 2.05 | 89 | | 125.0 | 143.6 | 148.0 | 97 |
| | Saliva 3<br>(0.23) | | 0.03 | 0.26 | 97 | Serum 3<br>(31.1) | 2.0 | 32.2 | 33.1 | 97 |
| | | | 0.06 | 0.27 | 92 | | 3.9 | 34.6 | 35.0 | 99 |
| | | | 0.13 | 0.39 | 108 | | 7.8 | 35.8 | 38.9 | 92 |
| | | | 0.25 | 0.50 | 103 | | 15.6 | 44.7 | 46.7 | 96 |
| | | | 0.50 | 0.84 | 114 | | 31.3 | 59.2 | 62.4 | 95 |
| | | | 1.00 | 1.48 | 120 | | 62.5 | 95.6 | 93.6 | 102 |
| | | | 2.00 | 2.65 | 119 | | 125.0 | 146.3 | 156.1 | 94 |
| Method<br>Comparison | Saliva | | | IBL-ELISA = 0.92 x IBL-Luminescence IA + 0.06 | | | | | r = 0.995; n = 130 | |
| | Serum | | | IBL-ELISA = 1.17 x IBL-Luminescence IA - 2.2 | | | | | r = 0.997; n = 129 | |
| | | | | IBL-ELISA = 0.97 x GCMS + 2.3 | | | | | r = 0.982; n = 33 | |

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Image /page/4/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle or bird-like symbol with three curved lines representing its body and wings. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the bird symbol.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Victor Herbst IBL-Hamburg Gmbh Flughafenstrasse 52a Hamburg, D-22335 Germany

Re:

DEC 2 0 2006

k062626 Trade/Device Name: Cortisol ELISA test kit Regulation Number: 21 CFR 862.1205 Regulation Name: Cortisol (Hydrocortisone and Hydroxycortisone) test Regulatory Class: Class II Product Code: CGR Dated: November 28, 2006 Received: November 30, 2006

Dear Mr. Herbst:

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.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21. Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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Page 2 -

This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97), You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours,

Jean M, Cooper MS, DVM

Jean M. Cooper, MS. D.V.M. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

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# Indications for Use

## 510(k) Number (if known):

K062626

Device Name: Cortisol ELISA test kit

Indications For Use:

The IBL Cortisol enzym linked immunosorbent assay is for the in-vitro-diagnostic quantitative determination of cortisol in human serum and saliva.

The Cortisol ELISA kit is useful as an aid in the differential diagnosis of Cushing syndrome and Addison's disease.

Prescription Use (Part 21 CFR 801 Subpart D) AND/OR

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

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Division Sign-Off

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Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) k062624

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