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.

Device Description

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 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.

AI/ML Overview

The provided text describes the Nichols Advantage® Chemiluminescence Erythropoietin Immunoassay. Here's a breakdown of the requested information based on the document:

Acceptance Criteria and Device Performance

The document does not explicitly state formal "acceptance criteria" with pass/fail thresholds. Instead, it compares the performance characteristics of the new device (Nichols Advantage™ EPO) with those of a legally marketed predicate device (Nichols Chemiluminescence EPO kit). The underlying assumption is that if the new device demonstrates comparable performance to the predicate, it meets the requirements for substantial equivalence.

Table of Performance Characteristics (Comparison to Predicate Device):

Feature	Acceptance Criteria (Implied by Predicate Performance)	Reported Device Performance (Nichols Advantage™ EPO)
Precision (Within-Run)	%CV comparable to predicate device	Mean (mU/mL): 12.5, %CV: 10.4
		Mean (mU/mL): 92, %CV: 3.3
		Mean (mU/mL): 168, %CV: 2.6
		Mean (mU/mL): 502, %CV: 4.5
Precision (Total)	%CV comparable to predicate device	Mean (mU/mL): 12.5, %CV: 13.6
		Mean (mU/mL): 92, %CV: 5.5
		Mean (mU/mL): 168, %CV: 3.8
		Mean (mU/mL): 502, %CV: 5.6
Recovery	Value comparable to predicate device (93-120%)	87-111%
Parallelism	Value comparable to predicate device (84-108%)	91-116%
High Dose Hook	Value comparable to predicate device (50,000 mU/mL)	7,500 mU/mL
Specificity & Cross-Reactivity (Human Transferrin)	<0.001%	<0.001%
Specificity & Cross-Reactivity (Human α-2-Macroglobulin)	<0.001%	<0.001%
Specificity & Cross-Reactivity (Human α-1-Acid Glycoprotein)	<0.001%	<0.001%
Specificity & Cross-Reactivity (Human α-1-Antitryspin)	<0.001%	<0.001%
Interference (Hemoglobin)	no interference up to 25 mg/dL	no interference up to 500 mg/dL
Interference (Triglycerides)	no interference up to 1000 mg/dL	no interference up to 3000 mg/dL
Interference (Unconjugated Bilirubin)	no interference up to 100 mg/dL	no interference up to 20 mg/dL
Interference (Albumin)	no interference up to 5000 mg/dL	no interference
Interference (Gamma Globulin)	no interference up to 5000 mg/dL	no interference
Method Comparison (Correlation)	R value comparable to predicate, indicating linearity	R = 0.960
Method Comparison (Range of Results)	Comparable range	14.7 to 590 mU/mL

Study Details:

Sample sizes used for the test set and the data provenance:
- Precision (Test Set):
  - Within-Run: 66 replicates for each of 4 concentration levels.
  - Total Precision: 66 replicates for each of 4 concentration levels.
- Method Comparison (Test Set): n=88 samples.
- Data Provenance: Not explicitly stated (e.g., country of origin). The study seems to be conducted by the manufacturer, Nichols Institute Diagnostics. It is a prospective study given the device is new and being compared to an existing one.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- This is an in vitro diagnostic (IVD) assay for quantitative determination of erythropoietin. The "ground truth" for such assays is typically established by reference methods or gravimetric/volumetric standards, not human expert consensus. The document does not describe expert involvement for ground truth establishment.
Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- Not applicable. This is an IVD assay, not an imaging or clinical decision support device requiring adjudication of expert interpretations.
If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- Not applicable. This is an IVD assay and does not involve human readers for interpretation in the context of an MRMC study.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- Yes, this is essentially a standalone performance evaluation of the immunoassay. The device performs the measurement and provides a quantitative result without human interpretative input beyond operating the system and performing quality control.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- For an immunoassay, the "ground truth" for performance evaluations is typically established against calibrated reference materials or a validated reference method.
  - The document states that both assays "are standardized to the same WHO Standard (1st IS 87/684) for recombinant DNA derived erythropoietin." This indicates that the ground truth for measuring erythropoietin levels is tied to this internationally recognized standard.
  - Precision and linearity are assessed against known concentrations (derived from standards).
  - Recovery measures the ability to detect known amounts of added analyte.
  - Method comparison uses patient samples, but the "ground truth" for comparison is the measurement obtained from the predicate device (which itself is standardized).
The sample size for the training set:
- This is an immunoassay, not a machine learning model that requires a distinct "training set" in the conventional sense. The calibration curve is established using known concentrations of recombinant EPO standards according to an instrument-specific process described (2-point calibration and master curve via reagent bar codes). No separate training set size is provided.
How the ground truth for the training set was established:
- As noted above, this device does not have a "training set" in the AI/ML context. The calibration ("training") of the assay itself is done using recombinant EPO as standards and standardized to the WHO Standard (1st IS 87/684). This ensures that the instrument's measurement of EPO concentrations is accurate and traceable to an accepted reference.

Summary

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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: (714) 240-5260 (714) 240-5313 Fax:

Summary Prepared On: March 18, 1999

1. Device Name;
  Trade/Proprietary Name: Nichols Advantage® Chemiluminescence Erythropoletin Immunoassav Erythropoietin Assay Common/Usual Name: Ervthropoletin Assay Classification Name:
1. 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.

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

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™Immunoassay	Nichols Institute DiagnosticsChemiluminescence EPO kit
Sample Size	200 uL	100 uL
Calibration	Full calibration curve every weekwith 2 point calibration every 4hours.	Full standard curve with every assayrun.
Solid Phase	Streptavidin coated magneticparticles.	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 roomtemperature (15-30°C)
Assay Counting	Automated on-board instrument	Semi-automated luminometer.
Assay Washing	Automated on-board instrument	Separate semi-automated washinstrument.

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Performance Characteristics:					Nichols Chemiluminescence EPO
Feature	Nichols Advantage™ EPO			Nichols Assay kit.
	(NCCLS Protocol)
Precision	Mean(mU/mL)	No ofReps.	%CV	Mean(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*:Human Transferrin @10 mg/mLHuman α-2-Macroglobulin @ 8.3 mg/mLHuman α-1-Acid Glycoprotein @8.3 mg/mLHuman α-1-Antitryspin @10 mg/mL	<0.001%<0.001%<0.001%<0.001%			<0.001%<0.001%<0.001%<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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Page 2

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 (770) 488-7655.

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 (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. 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 (800) 638-2041 or (301) 443-6597, 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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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:

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

Regulation Number and Section

§ 864.7250 Erythropoietin assay.

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