ELECSYS ESTRADIOL ASSAY

K965109 · Boehringer Mannheim Corp. · CHP · Feb 18, 1997 · Clinical Chemistry

Device Facts

• Record ID: K965109
• Device Name: ELECSYS ESTRADIOL ASSAY
• Applicant: Boehringer Mannheim Corp.
• Product Code: CHP · Clinical Chemistry
• Decision Date: Feb 18, 1997
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1260
• Device Class: Class 1

Intended Use

Immunoassay for the in vitro quantitative determination of estradiol in human serum and plasma.

Device Story

Elecsys® Estradiol Assay is an in vitro diagnostic immunoassay for quantitative estradiol measurement in human serum and plasma. Device uses competition principle; 50 µL sample incubated with biotinylated estradiol-specific antibody; streptavidin-coated microparticles and ruthenium-labeled estradiol derivative added. Microparticles magnetically captured on electrode surface; voltage application induces chemiluminescent emission measured by photomultiplier. Results calculated via instrument-specific calibration curve and reagent bar code master curve. Used in clinical laboratory settings on Elecsys® 2010 instrument. Provides quantitative estradiol levels to assist clinicians in endocrine assessment and patient management.

Clinical Evidence

Bench testing only. Precision evaluated using modified NCCLS protocols (N=60 per level); Elecsys showed improved precision (Total %CV 5.0-9.0%) compared to predicate (Total %CV 9.1-16.1%). Linearity established from 10-4600 pg/mL. Method comparison against predicate (N=74) yielded y=0.902x + 42.94, r=0.976. Interference testing performed for bilirubin, hemoglobin, lipemia, and biotin. Specificity cross-reactivity profiles provided for various steroids.

Technological Characteristics

Electrochemiluminescence immunoassay; uses streptavidin-coated microparticles and ruthenium complex (Tris(2,2'-bipyridyl)ruthenium(II)) label. Requires Elecsys® 2010 instrument. Calibration via 2-point calibration and reagent bar code master curve. Assay duration 18 minutes at 37 °C.

Indications for Use

Indicated for the in vitro quantitative determination of estradiol in human serum and plasma for clinical diagnostic purposes.

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.

Predicate Devices

• Enzymun-Test® Estradiol Assay (K916132)

Submission Summary (Full Text)

{0}

FEB 18 1997
K9105109

BOEHRINGER MANNHEIM CORPORATION
510(k) Summary

Introduction
According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

1. Submitter name, address, contact
Boehringer Mannheim Corporation
2400 Bisso Lane
P.O. Box 4117
Concord, CA 94524-4117
(510) 674 - 0690 extension 8413
FAX (510) 687 - 1850
Contact Person: Yvette Lloyd
Date Prepared: December 13, 1996

2. Device name
Proprietary name: Elecsys® Estradiol Assay
Common name: Electrochemiluminescence assay for the determination of estradiol.
Classification name: System, Test, Estradiol

3. Predicate device
We claim substantial equivalence to the Enzymun-Test® Estradiol Assay (K916132).

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510(k) Summary, Continued

## 4. Device Description

Competition principle. Total duration of assay: 18 minutes, 37 °C.

- 1st incubation (9 minutes): By incubating the sample (50 µL) with an estradiol-specific biotinylated antibody (65 µL), an immunocomplex is formed, the amount of which is dependent upon the analyte concentration in the sample.
- 2nd incubation (9 minutes): After addition of streptavidin-coated microparticles (35 µL) and an estradiol derivative labeled with a ruthenium complex** (65 µL), the still-vacant sites of the biotinylated antibodies become occupied, with the formation of an antibody-hapten complex. The entire complex becomes bound to the solid phase via interaction of biotin and streptavidin.
- The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier (0.4 second read frame).
- Results are determined via a calibration curve which is instrument-specifically generated by 2-point calibration and a master curve provided via the reagent bar code.

**Tris(2,2'-bipyridyl)ruthenium(II) complex (Ru(bpy)³⁺)

## 5. Intended use

Immunoassay for the in vitro quantitative determination of estradiol in human serum and plasma.

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510(k) Summary, Continued

6. Comparison to predicate device

The Boehringer Mannheim Elecsys® Estradiol Assay is substantially equivalent to other products in commercial distribution intended for similar use. Most notably it is substantially equivalent to the currently marketed Enzymun-Test® Estradiol Assay (K916132).

The following table compares the Elecsys® Estradiol Assay with the predicate device, Enzymun-Test® Estradiol Assay. Specific data on the performance of the test have been incorporated into the draft labeling in attachment 5. Labeling for the predicate device in provided in attachment 6.

Similarities:

- Intended Use: Immunoassay for the in vitro quantitative determination of estradiol
- Sample type: Serum and plasma
- Antibody: Same polyclonal anti-estradiol antibody
- Solid phase binding principle: Streptavidin/Biotin
- Assay Standardization: ID-GC/MS

Differences:

| Feature | Elecsys® Estradiol | Enzymun® Estradiol |
| --- | --- | --- |
| Reaction test principle | Electrochemiluminescence | ELISA/1-step sandwich assay. |
| Instrument required | Elecsys® 2010 | ES 300 |
| Calibration Stability | A calibration is recommended every 7 days if kit is not consumed; 4 weeks with same reagent lot if reagent is consumed within 7 days. | Full calibration required every 2 weeks. One-point calibration required every run. |

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510(k) Summary, Continued

6. Comparison to predicate device, cont.

Performance Characteristics:

| Feature | Elecsys® Estradiol | | | Enzymun® Estradiol | | |
| --- | --- | --- | --- | --- | --- | --- |
| Precision | Modified NCCLS (pg/mL): | | | Modified NCCLS (pg/mL): | | |
| Level | Low | Mid | High | Low | Mid | High |
| N | 60 | 60 | 60 | 60 | 60 | 60 |
| Within-Run: Mean | 53.3 | 410 | 3021 | 44.0 | 277.6 | 1217.0 |
| %CV | 6.5 | 2.7 | 3.0 | 14.3 | 8.5 | 7.7 |
| Total: Mean | 53.3 | 410 | 3021 | 44.0 | 277.6 | 1217.0 |
| %CV | 9.0 | 5.0 | 5.5 | 16.1 | 9.1 | 10.3 |
| Lower Detection Limit | 10 pg/mL | | | 10 pg/mL | | |
| Linearity | 10-4600 pg/mL (with a deviation from a linear line of ±10%) | | | 10-1300 pg/mL (with a deviation from a linear line of ±10%) | | |
| Method Comparison | Vs Enzymun-Test® Estradiol
Least Squares
y = 0.902x + 42.94
r = 0.976
SEE = 91.96
N = 74

Passing/Bablok
y = 0.962x + 12.51
r = 0.976
SEE = 38.71
N = 74 | | | Vs Enzymun-Test® Estradiol
Least Squares
y = 1.027x + 6.32
r = 0.995
SEE = 31.03
N = 64 | | |
| Interfering substances
Bilirubin
Hemoglobin
Lipemia
Biotin | No interference at:
25 mg/dL
1.0 g/dL
1500 mg/dL
24 ng/mL | | | No interference at:
64.5 mg/dL
200 mg/dL
1250 mg/dL
20 ng/mL | | |

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# 510(k) Summary, Continued

## 6. Comparison to predicate device, cont.

### Performance Characteristics:

| Feature | Elecsys® Estradiol | | Enzymum® Estradiol | |
| --- | --- | --- | --- | --- |
| Specificity | Level tested (ng/mL) | % Cross-reactivity | Level tested | % Cross-reactivity |
| 17 hydroxy-Progesterone | 100 | < 0.5 | --- | < 0.001 |
| Ethisterone | 100 | < 0.5 | --- | --- |
| Norethindrone-Acetate | 100 | < 0.5 | --- | --- |
| 2-Methoxy-estradiol | 100 | < 1.0 | --- | 0.07 |
| 6-Hydroxy-estradiol | 0.78 | < 600 | --- | 62 |
| Estriol | 160 | < 1.0 | --- | 0.3 |
| Prednisolone | 1000 | < 0.25 | --- | --- |
| Danazol | 100000 | <0.1 | --- | 0.0006 |
| Testosterone | 100000 | <1.0 | --- | 0.03 |
| Di-Hydro-Testosterone | 100000 | < 1.0 | --- | --- |
| 5-Androsten-3β-17β-Diol | 100000 | < 0.4 | --- | --- |