ADVANTAGE CHEMILUMINESCENCE ERYTHROPOIETIN IMMUNOASSAY

K980737 · Nichols Institute Diagnostics · GGT · Mar 23, 1999 · Hematology

Device Facts

• Record ID: K980737
• Device Name: ADVANTAGE CHEMILUMINESCENCE ERYTHROPOIETIN IMMUNOASSAY
• Applicant: Nichols Institute Diagnostics
• Product Code: GGT · Hematology
• Decision Date: Mar 23, 1999
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7250
• Device Class: Class 2

Indications for Use

The Advantage Chemiluminescence Erythropoietin Immunoassay is intended for use with the Advantage Specially System for the quantitative determination of erythropoietin concentrations in human serum as an adjunct in the diagnosis of anemia and polycythemia.

Device Story

The Nichols Advantage™ EPO assay is a two-site chemiluminescence immunoassay performed on the Advantage Specialty System. Input: 200uL human serum sample. Process: Sample incubated with biotinylated polyclonal antibody and acridinium-labeled mouse monoclonal antibody to form a sandwich complex; streptavidin-coated magnetic particles added to capture complex; automated washing and magnetic separation performed on-board. Trigger solutions (hydrogen peroxide/acid and sodium hydroxide) injected to oxidize acridinium ester, emitting light. Output: Light emission quantified in relative light units (RLU) by integrated luminometer, proportional to EPO concentration. Results calculated via instrument-generated calibration curve using reagent bar codes. Used in clinical laboratory settings by trained personnel. Provides quantitative EPO levels to assist clinicians in diagnosing anemia and polycythemia.

Clinical Evidence

Bench testing only. Performance evaluated using NCCLS protocols. Precision: within-run CV 2.6-10.4%, total precision CV 3.8-13.6%. Recovery: 87-111%. Parallelism: 91-116%. High dose hook: 7,500 mU/mL. Specificity: no significant cross-reactivity with transferrin, alpha-2-macroglobulin, alpha-1-acid glycoprotein, or alpha-1-antitrypsin. Interference testing: no interference from hemoglobin (up to 500 mg/dL), triglycerides (up to 3000 mg/dL), or unconjugated bilirubin (up to 20 mg/dL). Method comparison (n=88) against predicate showed Deming linear regression y = 0.736(x) + 10.9 and correlation R=0.960.

Technological Characteristics

Two-site chemiluminescence immunoassay. Solid phase: streptavidin-coated magnetic particles. Labels: biotinylated polyclonal antibody and acridinium-labeled mouse monoclonal antibody. Energy source: automated on-board luminometer. Connectivity: reagent bar code system for master curve data. Dimensions/Form factor: cuvette strip-based reaction wells. Software: integrated system for automated pipetting, incubation, washing, and signal quantification. Standardized to WHO Standard 1st IS 87/684.

Indications for Use

Indicated for quantitative determination of erythropoietin (EPO) concentrations in human serum as an adjunct in the diagnosis of anemia and polycythemia. For prescription use.

Regulatory Classification

Identification

A erythropoietin assay is a device that measures the concentration of erythropoietin (an enzyme that regulates the production of red blood cells) in serum or urine. This assay provides diagnostic information for the evaluation of erythrocytosis (increased total red cell mass) and anemia.

Special Controls

*Classification.* Class II. The special control for this device is FDA's “Document for Special Controls for Erythropoietin Assay Premarket Notification (510(k)s).”

Predicate Devices

• Nichols Institute Diagnostics Chemiluminescence Erythropoietin Immunoassay Kit (K952559)

Submission Summary (Full Text)

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3/22/99

# 510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K980737

- Name of Submitter, Contact Person and Date Summary Prepared: 1.
Jimmy Wong Nichols Institute Diagnostics 33051 Calle Aviador San Juan Capistrano, California 92675 Phone: [phone] [phone] Fax:

Summary Prepared On: March 18, 1999

- 2. Device Name;
Trade/Proprietary Name: Nichols Advantage® Chemiluminescence Erythropoletin Immunoassav Erythropoietin Assay Common/Usual Name: Ervthropoletin Assay Classification Name:

- 3. Legally Marketed Equivalent Device Name:
We are claiming substantial equivalence to the assay cleared in Nichols Institute Diagnostics' 510(k) K952559 for the Chemiluminescence Erythropoletin Immunoassay Kit.

- Description of the Device: 4.
The Nichols Advantage™ EPO assay is a two-site chemiluminescence assay. Total assay duration is 30 minutes at 37°C.

14 Incubation: 20 minutes at 37℃. Sample or control (200uL), biotinylated polyclonal antibody (40uL), acridinium labeled mouse monoclonal antibody (10uL) are pipetted into a reaction well on the cuvette strip. Each antibody binds to a separate and distinct antigenic site on EPO to form a sandwich complex.

204 Incubation: 10 minutes at 37°C. Streptavidin coated magnetic particles (20uL) are added to the reaction mixture. After the 10 minute incubation, the sandwich complex is bound to the solid phase via the high affinity interaction of blotin and streptavidin. The reaction mixture is aspirated from the reaction well after the streptavidin magnetic particles are magnetically capfured onto the surface of the reaction well wall.

Acridinum esters emit light upon treatment with hydrogen peroxide and an alkaline solution. The Trigger 1 solution contains hydrogen peroxide in dilute acid and Trigger 2 solution contains sodium hydroxide. The system automatically injects Trigger 1 and 2 into the reaction well which oxidizes the acridinium ester. The oxidized product is in an excited state. The subsequent return to ground state results in the emission of light which

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is quantified in 2 seconds, and is expressed in relative light units (RLU) by the integrated system luminometer.

The amount of bound labeled antibody in RLU's is directly proportional to the concentration of EPO in the sample. 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 codes.

#### ર્ડ Intended Use of the Device

The Advantage Chemiluminescence Erythropoietin Immunoassay is intended for use with the Advantage Specially System for the quantitative determination of erythropoietin concentrations in human serum as an adjunct in the diagnosis of anemia and polycythemia.

### Comparison of technological characteristics With Predicate Davice ર.

The Advantage Chemiluminescence Erythropoletin (EPO) Immunoassay is substantially equivalent in intended use to results obtained using the predicate device. Both assays use a labeled antibody for the quantitative measurement of EPO in serum. The major difference in the assays is that the subject device uses streptavidin coated magnetic particle In the capture separation phase of the assay; whereas, the predicate device uses an avidin coated bead as the separation system.

Companson of Performance Characteristics With Predicate Device 7.

> The Nichols Advantage™ EPO is substantially equivalent to other products in commercial distribution for similar use. The following tables compare the Nichols Advantage™ EPO with the predicate device, Nichols Chemiluminescence EPO kit.

## Similarities:

- Intended Use: For the quantitative determination of erythropoletin levels in human serum as . an adjunct in the diagnosis of anemia and polycythemia.
- Both assays use specific antibodies to EPO. .
- Both assays use human serum for the test sample. ●
- . The sensitivity of both assays are nearly the same, and both assays are standardized to the same WHO Standard (1ª IS 87/684) for recombinant DNA derived erythropoietin.
- Recombinant EPO is used as standards in both assays. .
- The upper end of the normal range is similar at the 95% confidence level.

## Differences:

| Feature | Nichols Advantage™<br>Immunoassay | Nichols Institute Diagnostics<br>Chemiluminescence EPO kit |
|----------------|---------------------------------------------------------------------------------|------------------------------------------------------------|
| Sample Size | 200 uL | 100 uL |
| Calibration | Full calibration curve every week<br>with 2 point calibration every 4<br>hours. | Full standard curve with every assay<br>run. |
| Solid Phase | Streptavidin coated magnetic<br>particles. | Avidin coated bead. |
| Dynamic Range | 5-700 mU/mL | 1.4-1500 mU/mL |
| Incubation | Total of 30 minutes at 37°C | Total of 4 hours at room<br>temperature (15-30°C) |
| Assay Counting | Automated on-board instrument | Semi-automated luminometer. |
| Assay Washing | Automated on-board instrument | Separate semi-automated wash<br>instrument. |

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| Performance Characteristics: | | | | | Nichols Chemiluminescence EPO | | |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------|----------------|------|------------------------------------------|-------------------------------|-----|--|
| Feature | Nichols Advantage™ EPO | | | Nichols Assay kit. | | | |
| | (NCCLS Protocol) | | | | | | |
| Precision | Mean<br>(mU/mL) | No of<br>Reps. | %CV | Mean<br>(mU/mL) | N | %CV | |
| Within-Run | 12.5 | 66 | 10.4 | 19.8 | 20 | 5.4 | |
| | 92 | 66 | 3.3 | 152 | 20 | 3.2 | |
| | 168 | 66 | 2.6 | 310 | 20 | 2.4 | |
| | 502 | 66 | 4.5 | 544 | 20 | 2.7 | |
| Total Precision | 12.5 | 66 | 13.6 | 21.8 | 20 | 9.7 | |
| | 92 | 66 | 5.5 | 160 | 20 | 6.1 | |
| | 168 | 66 | 3.8 | 294 | 20 | 5.3 | |
| | 502 | 66 | 5.6 | 544 | 20 | 3.7 | |
| Recovery | 87-111% | | | 93-120% | | | |
| Parallelism | 91-116% | | | 84-108% | | | |
| High Dose Hook | 7,500 mU/mL | | | 50,000 mU/mL | | | |
| Specificity & Crossreactivity*:<br>Human Transferrin @10 mg/mL<br>Human α-2-Macroglobulin @ 8.3 mg/mL<br>Human α-1-Acid Glycoprotein @8.3 mg/mL<br>Human α-1-Antitryspin @10 mg/mL | <0.001%<br><0.001%<br><0.001%<br><0.001% | | | <0.001%<br><0.001%<br><0.001%<br><0.001% | | | |
| Hemoglobin | no interference up to 500 mg/dL | | | no interference up to 25 mg/dL | | | |
| Triglycerides | no interference up to 3000 mg/dL | | | no interference up to 1000 mg/dL | | | |
| Unconjugated Bilirubin | no interference up to 20 mg/dL | | | no interference up to 100 mg/dL | | | |
| Albumin | no interference | | | no interference up to 5000 mg/dL | | | |
| Gamma Globulin | no interference | | | no interference up to 5000 mg/dL | | | |
| Method Comparison: | n=88 | | | | | | |
| Range of Results | 14.7 to 590 mU/mL | | | 11.4 to 702 mU/mL | | | |
| Deming Linear Regression | y = 0.736(x) + 10.9 | | | | | | |
| Correlation (R) | 0.960 | | | | | | |

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* Highest concentration tested with no interference.

1000 Children

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Public Health Service

Image /page/3/Picture/2 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is circular and contains the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" around the perimeter. Inside the circle is a stylized image of a human figure, represented by three overlapping profiles, with a bird-like shape above them.

# MAR 2 3 1999

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Jimmy Wonq Manager, Clinical and Technical Affairs Nichols Institute Diagnostics 33051 Calle Aviador San Juan Capistrano, CA 92675

Re: K980737

> Trade Name: Nichols Advantage® Chemiluminescence Erythropoietin Immunoassay Requlatory Class: III Product Code: GGT December 23, 1998 Dated: December 28, 1998 Received:

Dear Mr. Wong:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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 aqainst misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. this response to your premarket notification Please note: submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or requlations.

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Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at [phone].

This letter will allow you to begin marketing your device as described in your 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 advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification"(21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number [phone] or [phone], or at its internet address "http://www.fda.gov/cdrl/dsma/dsmamain.html".

Sincerely yours,

Steven Sutman

Steven I. Gutman, M.D. M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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7

## Indications For Use Statement 4.0

# INDICATIONS FOR USE STATEMENT

510(k) Number (if known): ____________________________________________________________________________________________________________________________________________________

Device Name: Nichols Advantage™ Chemiluminescence Erythropoietin Immunoassay

Indications For Use:

The Nichols Advantage™ Chemiluminescence Erythropoietin Immunoassay is intended for use with the Nichols Advantage™ Specialty System for the quantitative determination of erythropoietin concentrations in human serum as an adjunct in the diagnosis of anemia and polycythemia.

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of Device Evaluation (ODE)

| (Division Sign-Off) | |
|-----------------------------------------|---------|
| Division of Clinical Laboratory Devices | |
| 510(k) Number | 1980739 |

| Prescription Use (Per 21 CFR 801.109) | <div>✓</div> |
|---------------------------------------|--------------|
|---------------------------------------|--------------|

OR

| Over-The-Counter Use (Optional Format 1-2-96) |
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Febraury 18, 1998