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 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.

AI/ML Overview

The IBL Estradiol LIA is a luminescence immunoassay for the in vitro diagnostic quantitative measurement of active free Estradiol in saliva and serum.

1. Acceptance Criteria and Reported Device Performance

The acceptance criteria for this device are not explicitly stated as distinct pass/fail thresholds in the provided document. Instead, the document presents performance characteristics and comparisons to established methods. The "acceptance" is implied by the FDA's substantial equivalence determination. We can infer the functional performance criteria from the reported values and comparisons.

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance
Normal Ranges in Saliva	Establish distinct ranges for different physiological states.	Premenopausal (n=28): Follicular phase 0.6 - 10.4 pg/mL; Mid-cycle Peak 4.5 - 21.2 pg/mL; Luteal phase 0.5 - 10.8 pg/mL.Postmenopausal (n=5): < 3.2 pg/mL. Males (n=40): < 3.4 pg/mL.
Comparison to Published Saliva RIA	High correlation and close agreement with established methods.	r² = 0.97 with a regression formula of *Y = 0.96 RIA + 0.55** (n=50 saliva samples). Range measured: 0-51 pg/mL (RIA), 0-55 pg/mL (LIA).
Comparison to Commercial Serum RIA	High correlation and close agreement with established methods.	r² = 0.95 with a regression formula of *Y = 1.00 RIA - 4.83** (n=60 serum samples). Range measured: 22 - 448 pg/mL (RIA), 2 - 444 pg/mL (LIA).
Serum to Saliva Comparison	Demonstrate a relationship between serum and saliva levels.	R² = 0.712 with a regression formula of *serum (pg/mL) = 43.491 (saliva) - 4.680**.
Cross-Reactivity	Minimal cross-reactivity with structurally similar or common interfering substances.	17β-Estradiol: 100.0%Estrone: 0.222%Estriol: 0.138%Other substances (Corticosterone, Dexamethasone, Cortisone, Progesterone, Testosterone, Prednisolone): ≤ 0.01%
Analytical Sensitivity (LoD)	Low limit of detection for accurate measurement of low concentrations.	Saliva: 0.3 pg/mL (Mean signal (Zero-Standard) - 2SD)Serum: 7.3 pg/mL (Mean signal (Zero-Standard) - 2SD)
Functional Sensitivity	Concentration at which CV is <20%.	Saliva: 0.78 pg/mL (Mean Conc. <20% CV)Serum: 8.0 pg/mL (Mean Conc. <20% CV)
Precision (Intra-Assay & Inter-Assay)	Low coefficient of variation (CV) at different concentrations.	Saliva Intra-Assay: CVs from 3.7% to 7.9% (n=10)Saliva Inter-Assay: CVs from 5.9% to 13.9% (n=10)Serum Intra-Assay: CVs from 3.1% to 7.4% (n=10)Serum Inter-Assay: CVs from 4.3% to 9.3% (n=10)
Linearity	Measured values should be proportional to expected concentrations across a range of dilutions.	Saliva: Recoveries from 80% to 114% for various dilutions.Serum: Recoveries from 83% to 117% for various dilutions.
Recovery	Measured values should be close to added values.	Saliva: Recoveries from 92% to 128% for various spiked concentrations in 3 different saliva samples.Serum: Recoveries from 80% to 96% for various spiked concentrations in 3 different serum samples.

2. Sample Sizes Used for the Test Set and Data Provenance

Normal Ranges in Saliva:
- Premenopausal Women: 28 women (month profiles, so multiple samples per woman), age 19-43.
- Postmenopausal Women: 5 women, age 42-62.
- Males: 40 males, age 20-63.
- Data Provenance: Not explicitly stated, but likely prospective for the normal range establishment as saliva samples were collected daily "until first day of bleeding" or as single samples for postmenopausal women and males. No country of origin is specified, but the manufacturer is based in Germany, suggesting European or international data.
Comparison of Serum and Saliva (to published/commercial RIA):
- Saliva Comparison: 50 saliva samples.
- Serum Comparison: 60 serum samples.
- Data Provenance: Not explicitly stated, but likely retrospective or a combination of prospective/retrospective for convenience from adult healthy individuals. No country of origin is specified.
Serum to Saliva Comparison (using IBL Estradiol LIA):
- Paired Samples: Not explicitly stated, but derived from "saliva and serum pairs collected at the same time." The number of pairs is not provided.
- Data Provenance: Not explicitly stated, but likely prospective for this specific comparison.
Cross-Reactivity, Analytical Sensitivity, Functional Sensitivity, Precision, Linearity, Recovery:
- The sample sizes for these analytical performance studies are embedded within the tables (e.g., n=10 for precision at each level). These are typically laboratory studies using spiked samples, controls, and calibration materials rather than patient samples for the direct "test set" definition often associated with diagnostic device clinical trials.
- Data Provenance: Laboratory validation studies.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

Normal Ranges: The "ground truth" here is the observed ranges in the healthy populations studied. There is no mention of external experts establishing a ground truth for these ranges; they are empirical observations from the study participants themselves.
Comparison Studies: The "ground truth" for the comparison studies are the results obtained from the "published procedure that used a modification in the handling of saliva for a typical RIA test" and a "commercially available RIA test kit." These are established reference methods. There is no mention of individual expert adjudicators for these data points.

4. Adjudication Method for the Test Set

Not applicable in the context of this device. The comparisons are against established laboratory methods, not subjective diagnoses requiring adjudication by human experts.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

Not applicable. This is an in vitro diagnostic immunoassay, not an imaging or diagnostic device requiring interpretation by human readers. Therefore, an MRMC study and effects of AI assistance are not relevant to this submission.

6. Standalone Performance Study

Yes, the entire submission describes the standalone performance of the IBL Estradiol LIA. The comparisons to RIA methods ("Comparison of Serum and Saliva," "Serum to Saliva Comparison") are designed to demonstrate its performance relative to existing technologies, but the listed metrics (Normal Ranges, Cross-reactivity, Sensitivity, Precision, Linearity, Recovery) represent the intrinsic performance of the IBL Estradiol LIA device itself.

7. Type of Ground Truth Used

Normal Ranges: Empirical observation from defined healthy populations.
Comparison Studies: Results from established, commercially available, or published reference immunoassay methods (RIA).
Analytical Performance (Cross-Reactivity, Sensitivity, Precision, Linearity, Recovery): These are based on established laboratory analytical methods using controlled samples, calibrators, and known concentrations (e.g., spiked samples).

8. Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of machine learning or AI development. For an immunoassay, the equivalent would be the samples used to develop and optimize the assay's reagents, protocols, and calibration curves. This information is not provided in detail. The normal range studies and comparison studies would be considered part of the validation and testing, rather than a "training set" in the modern AI sense.

9. How the Ground Truth for the Training Set Was Established

As there is no distinct "training set" discussed in the context of AI/ML, this question is not directly applicable. However, for the development of the immunoassay itself, the "ground truth" for calibrators and controls would be established through rigorous analytical chemistry techniques, often traceable to international standards, and verified by expert analytical chemists during the assay development phase. The document implicitly relies on standard immunoassay development and validation practices.

Summary

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K041349

SUMMARY OF

SAFETY AND EFFECTIVENESS FOR IBL ESTRADIOL LIA

Manufacturer:	IBL Immuno Biological LaboratoriesFlughafenstrasse 52A, D-22335Hamburg Germany
Contact Information:	Lehnus & AssociatesGary Lehnus150 Cherry Lane Rd.East Stroudsburg, PA 18301Tel: (570) 620-0198

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:

Device Intended Use:

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)
♀	Premenopausaln=28 month profilesage 19 - 43	Follicular phase 0.6 - 10.4Mid-cycle Peak 4.5 - 21.2Luteal phase 0.5 - 10.8
♀	Postmenopausal n=5age 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 ofother substancestested:<= 0.01 %
Analytical Specificity(Cross Reactivity)	Corticosterone		0.007
Dexamethasone	0.009
Cortisone	0.007
Progesterone	0.012
Testosterone	0.015
Prednisolone	0.005
Analytical Sensitivity(Limit of Detection)	Saliva: 0.3 pg/mlSerum: 7.3 pg/mL	Mean signal (Zero-Standard) - 2SD
Functional Sensitivity	Saliva: 0.78 pg/mlSerum: 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.(pg/mL)	Rec.(%)	Dilution	Calculated (x10)(pg/mL)	Rec.(%)
	-	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(14.6)	0.9	15.2	98	Serum 1(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(5.5)	0.9	5.9	92	Serum 2(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(2.4)	0.9	3.8	113	Serum 3(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 toPublishedProcedure	Saliva	$IBL-Assay = 0.96 x RIA + 0.55$$R^2 = 0.97; n = 50$		Range = 0-51 pg/mL RIA0-55 pg/mL LIA
Comparison toCommercial RIA	Serum	$IBL-Assay = 1.00 x RIA - 4.83$$R^2 = 0.95; n = 60$		Range = 22 - 448 pg/mL RIA2 - 444 pg/mL LIA
Serum to SalivaComparison IBLLIA	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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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 (301) 594-3084. 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 (800) 638-2041 or (301) 443-6597 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:

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

Page 1 of

Regulation Number and Section

§ 862.1260 Estradiol test system.

(a)
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.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 862.9.