LUMINESCENT IMMUNOASSAY KIT FOR THE DETECTION OF ESTRADIOL IN SALIVA AND SERUM

K041349 · Ibl GmbH · CHP · Sep 24, 2004 · Clinical Chemistry

Device Facts

• Record ID: K041349
• Device Name: LUMINESCENT IMMUNOASSAY KIT FOR THE DETECTION OF ESTRADIOL IN SALIVA AND SERUM
• Applicant: Ibl GmbH
• Product Code: CHP · Clinical Chemistry
• Decision Date: Sep 24, 2004
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1260
• Device Class: Class 1

Intended Use

Luminescence immunoassay for the in vitro diagnostic quantitative measurement of active free estradiol, an estrogenic steroid, in saliva and serum. Measurements obtained by this device may be used in the diagnosis and treatment of various hormonal sexual disorders and can be used to evaluate ovarian function. This test is not intended for assessing placental function in complicated pregnancy.

Device Story

Luminescence immunoassay (LIA) for quantitative measurement of Estradiol in saliva and serum; utilizes competition principle where sample antigen and enzyme-labeled antigen compete for antibody binding sites on coated wells. Following incubation and wash steps, luminescence substrate is added; intensity of luminescence is inversely proportional to antigen concentration. Results determined via standard curve. Used in clinical laboratory settings by trained personnel. Output provides Estradiol concentration (pg/mL) to assist clinicians in diagnosing hormonal sexual disorders and evaluating ovarian function.

Clinical Evidence

Bench testing only. Precision/reproducibility evaluated with serum and saliva samples (CVs ≤7.9% within-run; ≤16.5% between-run). Linearity/recovery confirmed via serial dilution and spiking (recoveries 80-120%). Analytical sensitivity: 0.3 pg/mL (saliva), 0.7 pg/mL (serum). Specificity tested against various steroids; cross-reactivity ≤0.222%. Method comparison with predicate RIA (n=60 serum samples) showed y = 1.00x - 4.83, r² = 0.95. Comparison with published RIA for saliva (n=50) showed y = 0.96x + 0.55, r² = 0.97. Interference testing for triglycerides, bilirubin, hemoglobin, thimerosal, and sodium azide showed no significant effect.

Technological Characteristics

Microtiter plate coated with rabbit anti-mouse antibody; mouse anti-progesterone antiserum; alkaline phosphatase enzyme conjugate; acridan-based chemiluminescence substrate. Competitive binding immunoassay. Quantitative measurement via luminescence intensity. Standalone kit format. No specific connectivity or software algorithm class mentioned.

Indications for Use

Indicated for the quantitative measurement of active free estradiol in saliva and serum to assist in the diagnosis and treatment of hormonal sexual disorders and evaluation of ovarian function in patients. Not indicated for assessing placental function in complicated pregnancy.

Regulatory Classification

Identification

An estradiol test system is a device intended to measure estradiol, an estrogenic steroid, in plasma. Estradiol measurements are used in the diagnosis and treatment of various hormonal sexual disorders and in assessing placental function in complicated pregnancy.

Submission Summary (Full Text)

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K041349

#### SUMMARY OF

## SAFETY AND EFFECTIVENESS FOR IBL ESTRADIOL LIA

| Manufacturer: | IBL Immuno Biological Laboratories<br>Flughafenstrasse 52A, D-22335<br>Hamburg Germany |
|----------------------|------------------------------------------------------------------------------------------------------------------|
| Contact Information: | Lehnus & Associates<br>Gary Lehnus<br>150 Cherry Lane Rd.<br>East Stroudsburg, PA 18301<br>Tel: [phone] |

## Device Name / Classification:

The device trade name is the IBL Estradiol LIA having FDA assigned name: Estradiol test system, 21 CFR, 862.1260, categorized as Class 1 "exempt" medical devices for the Clinical Chemistry and Clinical Toxicology Panel, as Product Code CHP.

## Device Description:

Luminescence immunoassay (LIA) based on the competition principle. An unknown amount of antigen present in the sample and a fixed amount of enzyme labeled 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 addition of the luminescence substrate solution the intensity of the luminescence measured 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:

Luminescence immunoassay for the in vitro diagnostic quantitative measurement of active free Estradiol, an estrogenic steroid, in saliva and serum. Measurements obtained by this device may be used in the diagnosis and treatment of various hormonal sexual disorders and can be used to evaluate ovarian function. This test is not intended for assessing placental function in complicated pregnancy.

## Device Performance:

## Normal Ranges in Saliva

To establish a normal range in saliva for this test, studies were performed to establish levels of estradiol in saliva throughout the menstrual cycles from 28 apparently healthy pre-menopausal women using no contraceptives. Three saliva samples were collected per day (morning. midday, and afternoon) and pooled and frozen prior to running the Estradiol LIA assay. Collection began at the last day of bleeding and continued daily until first day of bleeding. In addition, five postmenopausal women and 40 males were evaluated for the studies. One saliva sample was obtained for these 2 populations. The Normal Ranges in saliva obtained for the studies are as follows:

| Normal ranges | Estradiol (pg/mL) | |
|---------------|-----------------------------------------------------|-------------------------------------------------------------------------------------|
| ♀ | Premenopausal<br>n=28 month profiles<br>age 19 - 43 | Follicular phase 0.6 - 10.4<br>Mid-cycle Peak 4.5 - 21.2<br>Luteal phase 0.5 - 10.8 |
| ♀ | Postmenopausal n=5<br>age 42 - 62 | < 3.2 |
| ♂ | n = 40, age 20 - 63 | < 3.4 |

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Comparison of Serum and Saliva:

Comparison studies were performed using 50 saliva and 60 serum samples from adult healthy Oompanion the saliva samples were tested on the IBL Estradiol LIA and compared to a published procedure that used a modification in the handling of saliva for a typical RIA test. Results from measuring the saliva samples in both methods yielded a correlation of r2 = 0.97 with a regression formula of Y=0.96°RIA + 0.55. The serum samples were tested against a with a regially available RIA test kit. The comparison with serum yielded r2 = 0.95 with a regression formula of Y= 1.00*RIA - 4.83.

Serum to Saliva Comparison:

Sertam to Galliva Compuniou.
A study was performed with the IBL Estradiol LIA to assess the levels of estradiol in serum versus those found in saliva. Saliva and serum pairs were collected at the same time between 10:00 AM and 4:00 PM. The paired samples were then run in the IBL Estradiol LIA test. The results of the comparison yielded the following regression formula - serum (pg/mL) = 43.491(saliva) - 4.680 with a R2 = 0.712.

| Substance | % Cross-reactivity | | | | | | | |
|------------------------------------------------|----------------------------------------|-----------------------------------------------------------------|-------------|---------------|-----------------------------|-------------|--------|----|
| 17β-Estradiol | 100.0 | | | | | | | |
| Estrone | 0.222 | | | | | | | |
| Estriol | 0.138 | Cross-reactivity of<br>other substances<br>tested:<br><= 0.01 % | | | | | | |
| Analytical Specificity<br>(Cross Reactivity) | Corticosterone | | 0.007 | | | | | |
| Dexamethasone | 0.009 | | | | | | | |
| Cortisone | 0.007 | | | | | | | |
| Progesterone | 0.012 | | | | | | | |
| Testosterone | 0.015 | | | | | | | |
| Prednisolone | 0.005 | | | | | | | |
| Analytical Sensitivity<br>(Limit of Detection) | Saliva: 0.3 pg/ml<br>Serum: 7.3 pg/mL | Mean signal (Zero-Standard) - 2SD | | | | | | |
| Functional Sensitivity | Saliva: 0.78 pg/ml<br>Serum: 8.0 pg/mL | Mean Conc. <20% CV | | | | | | |
| Precision | Saliva (pg/mL) | | | Serum (pg/mL) | | | | |
| | Mean | SD | CV (%) | N | Mean | SD | CV (%) | N |
| Intra-Assay | 3.5 | 0.3 | 7.9 | 10 | 37.5 | 3 | 7.4 | 10 |
| | 11.8 | 0.5 | 7.3 | 10 | 167 | 8 | 4.8 | 10 |
| | 33.0 | 1.2 | 3.7 | 10 | 334 | 10 | 3.1 | 10 |
| Inter-Assay | 2.4 | 0.3 | 13.9 | 10 | 34.1 | 3.2 | 9.3 | 10 |
| | 9.4 | 0.9 | 9.1 | 10 | 159.2 | 11.9 | 7.5 | 10 |
| | 38.5 | 2.2 | 5.9 | 10 | 329 | 14.1 | 4.3 | 10 |
| Linearity | Saliva | | | Serum | | | | |
| | Dilution | Meas.<br>(pg/mL) | Rec.<br>(%) | Dilution | Calculated (x10)<br>(pg/mL) | Rec.<br>(%) | | |
| | - | 8.0 | | - | 312 | | | |
| | 1:2 | 3.9 | 98 | 1:2 | 134 | 86 | | |
| | 1:4 | 2.2 | 111 | 1:4 | 65 | 83 | | |
| | 1:8 | 1.1 | 106 | 1:8 | 34 | 88 | | |
| | - | 15.5 | | 1:16 | 18 | 94 | | |
| | 1:2 | 7.9 | 102 | - | 178 | | | |
| | 1:4 | 4.1 | 106 | 1:2 | 72 | 81 | | |
| | 1:8 | 1.6 | 80 | 1:4 | 38 | 85 | | |
| | - | 49.6 | | 1:8 | 19 | 86 | | |

The overall performance of the IBL Estradiol LIA is shown following:

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| 1:2 | 24.4 | 98 | 1:16 | 13 | 117 | | |
|------------------------------------------|----------------------------------------|---------------------------------------------------------|--------|-------------------------------------------------|-------------|------------------------|--------|
| 1:4 | 11.3 | 91 | | 38 | | | |
| 1:8 | 7.1 | 114 | 1:2 | 19 | 100 | | |
| 1:16 | 3.4 | 111 | 1:4 | 8 | 87 | | |
| 1:32 | 1.4 | 90 | 1:8 | 5 | 98 | | |
| Conc. (pg/mL) | Added pg/mL | Meas. pg/mL | Rec. % | Conc. (pg/mL) | Added pg/mL | Calculated (x10) pg/mL | Rec. % |
| Saliva 1<br>(14.6) | 0.9 | 15.2 | 98 | Serum 1<br>(52) | 9 | 58 | 96 |
| | 2 | 15.9 | 95 | | 20 | 67 | 93 |
| | 4 | 18.8 | 101 | | 40 | 81 | 89 |
| | 8 | 24.5 | 109 | | 80 | 110 | 83 |
| | 16 | 28.7 | 94 | | 160 | 188 | 89 |
| | 32 | 56.9 | 122 | | 320 | 313 | 84 |
| Saliva 2<br>(5.5) | 0.9 | 5.9 | 92 | Serum 2<br>(221) | 9 | 207 | 90 |
| | 2 | 6.2 | 83 | | 20 | 229 | 95 |
| | 4 | 9.1 | 95 | | 40 | 230 | 88 |
| | 8 | 14.4 | 106 | | 80 | 257 | 85 |
| | 16 | 21.8 | 101 | | 160 | 335 | 88 |
| | 32 | 47.9 | 128 | | 320 | 434 | 80 |
| Saliva 3<br>(2.4) | 0.9 | 3.8 | 113 | Serum 3<br>(364) | 9 | 375 | 101 |
| | 2 | 5.2 | 116 | | 20 | 387 | 101 |
| | 4 | 6.7 | 105 | | 40 | 394 | 97 |
| | 8 | 11.1 | 106 | | 80 | 427 | 96 |
| | 16 | 20.3 | 110 | | 160 | 469 | 90 |
| | 32 | 41.2 | 120 | | 320 | 636 | 93 |
| Comparison to<br>Published<br>Procedure | Saliva | $IBL-Assay = 0.96 x RIA + 0.55$<br>$R^2 = 0.97; n = 50$ | | Range = 0-51 pg/mL RIA<br>0-55 pg/mL LIA | | | |
| Comparison to<br>Commercial RIA | Serum | $IBL-Assay = 1.00 x RIA - 4.83$<br>$R^2 = 0.95; n = 60$ | | Range = 22 - 448 pg/mL RIA<br>2 - 444 pg/mL LIA | | | |
| Serum to Saliva<br>Comparison IBL<br>LIA | Serum (pg/mL) = 43.491(saliva) - 4.680 | | | $R^2 = 0.712$ | | | |

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/3/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo features a stylized eagle with three stripes representing the three branches of government. The eagle is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES U.S.A."

SEP 2 4 2004

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

IBL Hamburg c/o Mr. Gary Lehnus Lehnus & Associates Consulting 150 Cherry Lane Rd. East Stroudsburg, PA 18301

Re: k041349

> Trade/Device Name: IBL Estradiol LIA Test Regulation Number: 21 CFR 862.1260 Regulation Name: Estradiol test system Regulatory Class: Class I Product Code: CHP Dated: September 3, 2004 Received: September 7, 2004

Dear Mr. Lehnus:

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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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

Enclosure

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# Indications for Use

510(k) Number (if known): K041349

Device Name: ________IBL Estradiol LIA Test

# Indications For Use:

Luminescence immunoassay for the in vitro diagnostic quantitative measurement of active free Estradiol, an estrogenic steroid, in saliva and serum. Measurements obtained by this device may be used in the diagnosis and treatment of various hormonal sexual disorders and can be used to evaluate ovarian function. This test is not intended for assessing placental function in complicated pregnancy.

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

Over-The-Counter Use (21 CFR 807 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

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) k041349

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