K Number

Device Name

SEALITE SCIENCES, INC. AQUALITE FERRITIN ASSAY

Manufacturer

SEALITE SCIENCES, INC.

Date Cleared

1997-06-24

(126 days)

Product Code

DBF

Regulation Number

866.5340

Intended Use

The AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or the AquaLite® Ferritin Assay) is an in vitro diagnostic product intended for use in clinical laboratories for the quantitative determination of human ferritin in serum and plasma. Ferritin measurements are used in the diagnosis of diseases affecting iron metabolism.

Device Description

The AquaLite® Ferritin Bioluminescent Immunoassay Kit uses a mouse monoclonal antiferritin antibody that is pre-coated onto polystyrene tubes (solid phase). Samples (serum or plasma), or appropriate calibrators or controls, are pipetted (25 uL) into the precoated tubes. A mouse monoclonal anti-ferritin antibody covalently linked to AquaLite® (150 uL) is then added to the tubes. Ferritin in the sample simultaneously combines with the antibody on the solid phase and conjugate antibody to form an immune complex or "sandwich" bound to the solid phase. Complex formation is complete after a 60-minute incubation period at room temperature on a standard orbital shaker. The tubes are then washed to remove unbound conjugate. The washed tubes are placed in a luminometer that is capable of reading a triggered, flash-type reaction in 12 x 75 mm tubes. Injection of the calcium trigger buffer causes AquaLite® to oxide its self-contained luciferin molecule, producing a flash of light, which is measured by the luminometer. The intensity of the light emitted from antibody bound to the tubes is directly proportional to the concentration of the ferritin in the sample. To calculate results, the light intensity (in relative light units, RLU) of the ferritin calibrators is plotted against ferritin concentration (in ng per mL) to yield a calibration curve. This curve is used to relate the light intensity generated from the samples and controls to ferritin concentration in ng/mL. Note: Samples that generate signals greater than the signal from the highest calibrator are off-scale. These samples must be diluted and re-assayed. Remember to multiply the results from diluted samples by the dilution factor used.

More Information

Contact

Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

State

Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

Chiron Diagnostics ACS:180 Ferritin Assay

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description outlines a standard bioluminescent immunoassay with a luminometer reading and calibration curve calculation. There is no mention of AI or ML in the device description, intended use, or performance studies.

Therapeutic

No
This device is an in vitro diagnostic product used for the quantitative determination of human ferritin in serum and plasma, which aids in the diagnosis of diseases. It does not directly treat or prevent a disease, nor does it restore, modify, or correct body function or structure.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the product is an "in vitro diagnostic product" and is "intended for use in clinical laboratories for the quantitative determination of human ferritin in serum and plasma. Ferritin measurements are used in the diagnosis of diseases affecting iron metabolism." This directly indicates its purpose in diagnosing diseases.

SaMD (Software as a Medical Device)

The device description clearly outlines a physical kit containing antibodies, tubes, and reagents, which are hardware components. While software is used for calculation and potentially controlling the luminometer, the core of the device is a physical immunoassay kit.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use/Indications for Use: The very first sentence explicitly states it is an "in vitro diagnostic product intended for use in clinical laboratories for the quantitative determination of human ferritin in serum and plasma." This directly aligns with the definition of an IVD.
Device Description: The description details a laboratory test performed on biological samples (serum and plasma) outside of the body to measure a specific analyte (ferritin). This is characteristic of an in vitro diagnostic test.
Intended User/Care Setting: It is intended for use in "clinical laboratories," which are the typical settings for performing IVD tests.
Performance Studies: The document includes a summary of performance studies (sensitivity, specificity, precision, method comparison, etc.) which are required for the validation and regulatory approval of IVD devices.
Predicate Device(s): The mention of a "Predicate Device(s)" (Chiron Diagnostics ACS:180 Ferritin Assay) is a strong indicator that this device is being submitted for regulatory review as an IVD, often through a 510(k) pathway where comparison to a legally marketed predicate device is required.

All of these elements clearly indicate that the AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit is an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Product codes

DBF

Device Description

The washed tubes are placed in a luminometer that is capable of reading a triggered, flash-type reaction in 12 x 75 mm tubes. Injection of the calcium trigger buffer causes AquaLite® to oxide its self-contained luciferin molecule, producing a flash of light, which is measured by the luminometer. The intensity of the light emitted from antibody bound to the tubes is directly proportional to the concentration of the ferritin in the sample. To calculate results, the light intensity (in relative light units, RLU) of the ferritin calibrators is plotted against ferritin concentration (in ng per mL) to yield a calibration curve. This curve is used to relate the light intensity generated from the samples and controls to ferritin concentration in ng/mL.

Note: Samples that generate signals greater than the signal from the highest calibrator are off-scale. These samples must be diluted and re-assayed. Remember to multiply the results from diluted samples by the dilution factor used.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

clinical laboratories

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Studies on the AquaLite® Ferritin Assay were conducted at SeaLite Sciences.

Sensitivity: The sensitivity or detection limit of the AquaLite® Ferritin Assay is 0.03 ne/mL.
Specificity: Cross reactivity of the AquaLite® Ferritin assay was determined by measuring the concentration of ferritin in 2 human serum samples spiked with 300 ng/mL liver ferritin.
High Dose Hook Effect: No high dose hook effect occurs prior to 10,000 ng/mL ferritin.
Precision:
(a) Intra-assay precision: Two serum commercial controls were assayed to determine intra-assay precision (Total N = 20 per concentration level).
(b) Inter-assay precision: Commercial controls were assayed in duplicate repetitively. Ten (10) assays were performed and a new standard curve was generated for each assay (Total n = 2 x 10 = 20).
Method Comparison: The AquaLite® Ferritin Assay was used to assay patient samples (N=99) that were previously assayed by a commercially available chemiluminometric immunoassay. A slope of 0.72 with a y-intercept of 15.9 was obtained. The correlation coefficient was 0.98.
Linearity and Nonparallelism: Three human serum samples were diluted using Calibrator A (0 ng/mL) and assayed in duplicate.
Spike and Recovery: Three normal human serum samples were diluted with 120 ng/mL human spleen ferritin (WHO 2nd IS 80/578). The spiked samples were assayed using the AquaLite® Ferritin Assay.
Recovery in Serum and Plasma: Blood samples from 6 normal subjects were prepared as sera and as EDTA plasma. Ferritin was quantified using the AquaLite® Ferritin Assay. The data demonstrate that there are no significant differences among serum and EDTA plasma.
Effect of Common Interferents: Pooled normal human serum was spiked with preparations of hemoglobin, bilirubin, and triglycerides. The data demonstrate that the AquaLite® Ferritin assay is not significantly affected by hemoglobin, bilirubin, or triglycerides at the levels tested.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Sensitivity: 0.03 ne/mL
Specificity: Two human serum samples spiked with 300 ng/mL liver ferritin showed cross-reactivities of 85% and 103%.
Precision:
Intra-assay %CV: 8.6, 3.6, 5.4.
Inter-assay %CV: 4.5%, 3.7%, 9.3%.
Method Comparison: Correlation coefficient = 0.98.
Recovery: Spike and recovery percentages ranged from 84% to 100%. Recovery in Serum and Plasma percentages ranged from 85% to 103%.
Effect of Common Interferents: Recovered ferritin as a percentage of expected for Hgb at 500 mg/dL was 98%; for Bilirbn at 20 mg/dL was 90%; for Trig at 3000 mg/dL was 99%.

Predicate Device(s)

Chiron Diagnostics ACS:180 Ferritin Assay

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.5340 Ferritin immunological test system.

(a)
Identification. A ferritin immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the ferritin (an iron-storing protein) in serum and other body fluids. Measurements of ferritin aid in the diagnosis of diseases affecting iron metabolism, such as hemochromatosis (iron overload) and iron deficiency amemia.(b)
Classification. Class II (performance standards).

Summary

K970612

JUN 24 1997

510(k) SUMMARY

I. GENERAL INFORMATION

Trade or proprietary name - SeaLite Sciences, Inc. AquaLite® Ferritin Assay

Common or usual name - Bioluminescent immunoassay (BIA)

Classification name - FDA has classified ferritin test systems intended for the measurement of ferritin in the diagnosis of diseases affecting iron metabolism, such as hemochromatosis (iron overload) and iron deficiency anemia as Class II devices. (21 C.F.R. § 866.5340)

| Submitter's Name and Address: | Cathryn C. Cambria
Director, Regulatory Affairs and Quality
Assurance
SeaLite Sciences, Inc.
3000 Northwoods Parkway
Suite 200
Norcross, GA 30071
(800) 874-4471, ext. 227 |
|-----------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Submission Date: | February 14, 1997 |
| Legally Marketed Device
To Which Claim Substantial
Equivalence: | Chiron Diagnostics ACS:180 Ferritin Assay |

INDICATIONS FOR USE II.

DEVICE DESCRIPTION III.

IV. SUMMARY OF STUDIES AND TECHNOLOGICAL CHARACTERISTICS

Studies on the AquaLite® Ferritin Assay were conducted at SeaLite Sciences. The results are summarized below:

Performance Characteristics

1. Sensitivity
  The sensitivity or detection limit of the AquaLite® Ferritin Assay is 0.03 ne/mL. Sensitivity is determined by adding the mean signal of 20 replicates of the zero level calibrator plus two (2) standard deviations above this mean. The ferritin concentration (ng/mL) associated with this calculated signal is defined as the sensitivity of the assay.

2. Specificity

The AquaLite® Ferritin Assay measures intact ferritin. Cross reactivity of the AquaLite® Ferritin assay was determined by measuring the concentration of ferritin in 2 human serum samples spiked with 300 ng/mL liver ferritin. Percent crossreactivity (%) is reported below:

| Endogenous
Ferritin
(ng/mL) | Ferritin
Expected
(ng/mL) | Ferritin
Observed
(ng/mL) | Cross-
Reactivity
(%) |
|-----------------------------------|---------------------------------|---------------------------------|-----------------------------|
| 79 | 379 | 323 | 85 |
| 139 | 439 | 451 | 103 |

High Dose Hook Effect 3.

No high dose hook effect occurs prior to 10,000 ng/mL ferritin.

4. Precision

(a) Two serum commercial controls containing Intra-assay precision. ferritin at the following concentrations were assayed to determine intraassay precision. (Total N = 20 per concentration level.)

Ferritin Level (ng/mL)	% CV
368.4	8.6
123.9	3.6
40.5	5.4

(b) Inter-assay precision. Commercial controls containing ferritin at the following concentrations were assayed in duplicate repetitively. Ten (10) assays were performed and a new standard curve was generated for each assay. The inter-assay precision observed for the solutions (Total n = 2 x 10 = 20) are shown below.

Ferritin Level (ng/mL)	% CV
50.9	4.5%
121.6	3.7%
363.2	9.3%

5. Method Comparison

The AquaLite® Ferritin Assay was used to assay patient samples (N=99) that were previously assayed by a commercially available chemiluminometric immunoassay. A slope of 0.72 with a y-intercept of 15.9 was obtained. The correlation coefficient was 0.98.

6. Linearity and Nonparallelism

Three human serum samples containing the levels of endogenous ferritin shown below were diluted as indicated using Calibrator A (0 ng/mL) and assayed in duplicate. All concentrations are in ng/mL.

| SAMPLE
ID | DILUTION
FACTOR | OBSERVED
(ng/mL) | FERRITIN
EXPECTED
(ng/mL) | RECOVERY
(%) |
|--------------|--------------------------------|--------------------------------|---------------------------------|-------------------------------|
| A | Undiluted
1:2
1:4
1:8 | 160.1
77.3
47.4
23.9 | --
80
40
20 | --
96.6
118.5
119.5 |
| B | Undiluted
1:2
1:4
1:8 | 294.1
160.9
84.4
45.1 | --
147
74
27 | --
109.5
114.1
121.9 |
| C | Undiluted
1:2
1:4
1:8 | 878
383.0
189.6
101.4 | --
439
219.5
109.8 | --
87.2
86.4
92.3 |

7. Spike and Recovery

Three normal human serum samples were diluted with 120 ng/mL human spleen ferritin (WHO 2nd IS 80/578). The spiked samples were assayed using the AquaLite® Ferritin Assay. All values are in ng/mL.

| Sample | Unspiked
(ng/mL) | Spike
Recovered
(ng/mL) | Spiked
Expected
(ng/mL) | % |
|--------|---------------------|-------------------------------|-------------------------------|-----|
| 1 | 1260 | 595 | 690 | 86 |
| 2 | 58 | 90 | 90 | 100 |
| 3 | 406 | 221 | 263 | 84 |

8. Recovery in Serum and Plasma

Blood samples from 6 normal subjects were prepared as sera (standard tubes) as Ferritin was quantified using the AquaLite® Ferritin well as EDTA plasma. Assay. Recovered ferritin was compared with the ferritin recovered in serum (standard technique). The data demonstrate that there are no significant differences among serum and EDTA plasma when using the AquaLite® Ferritin Assay.

| Sample | Serum
(ng/mL) | EDTA
(ng/ml) | % |
|--------|------------------|-----------------|-----|
| 1 | 63.9 | 62.0 | 97 |
| 2 | 44.1 | 38.7 | 88 |
| 3 | 62.8 | 53.6 | 85 |
| 4 | 109.6 | 110.9 | 101 |
| 5 | 105.2 | 108.8 | 103 |
| 6 | 63.3 | 62.6 | 99 |

9. Effect of Common Interferents

Pooled normal human serum was spiked with preparations of hemoglobin, bilirubin, and triglycerides to the levels shown below. Ferritin was quantified using the AquaLite® Ferritin assay. Recovered ferritin was compared to the ferritin recovered in normal serum. The data demonstrate that the AquaLite® Ferritin assay is not significantly affected by hemoglobin, bilirubin, or triglycerides at the levels tested.

Analyte	Observed	Expected	%
Hgb at 500 mg/dL	113.3	115.6	98
Bilirbn at 20 mg/dL	114.2	115.6	90
Trig at 3000 mg/dL	116.2	128.9	99

V. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH

Use Universal Precautions. No known test method can offer complete Caution: assurance that products derived from human serum are pathogen-free; therefore, handle all materials of human origin as though they were potentially infectious.

Sodium azide is used as a preservative. This preservative may react with metallic plumbing to from explosive metal azides. Flush with large volumes of water when disposing of materials containing sodium azide.

D-6

As an in vitro diagnostic test, there are not direct adverse effects on the health of a patient from the use of this product. However, failure of the device to perform as indicated, the contamination of reagents, the use of reagents past the labeled expiration dates, the use of improper specimens, or human error during the performance of the test may lead to erroneous results and possible improper patient management.

VI. CONCLUSIONS DRAWN FROM STUDIES

The data from the studies conducted demonstrate that the performance of SeaLite Sciences, Inc. AquaLite® Ferritin Assay is similar and substantially equivalent to that of other commercially available assays for ferritin.

Image /page/7/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of an eagle with its wings spread, rendered in black lines. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" are arranged in a circular pattern around the eagle, also in black.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Cathryn C. Cambria Director, Regulatory Affairs & Quality Assurance SeaLite Sciences, Inc. 3000 Northwoods Parkway -----Suite 200 Norcross, GA 30071

JUN 2 4 1997

Re: K970612/S001 Trade Name: SeaLite Sciences, Inc. AquaLite® Ferritin Assay Regulatory Class: II Product Code: DBF Dated: May 08, 1997 Received: May 12, 1997

Dear Ms. Cambria:

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 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 against 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 Good Manufacturing Practice for Medical Devices: General (GMP) regulation (21 CFR Part 820) and that, through periodic GMP 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.

Please note: this response to your premarket notification 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 Regulations.

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.

Page 2

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 at (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Autman

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

Page 1 of _l

510(k) Number (if known): _

AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or the Device Name: AquaLite® Ferritin Assay)

Indications for Use:

The AquaLite® Ferritin Bioluminescent Immunoassay (BIA) Kit (or AquaLite® Ferritin Assay) is an in vitro diagnostic product intended for use in clinical laboratories for the quantitative measurement of ferritin in human serum and plasma. Ferritin measurements are used in the diagnosis of diseases affecting iron metabolism.

Peter E. Mafini

(PLEASE DO NOT WRITE BELOW THIS LINE CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use_V (Per 21 CFR 801.109)

Over-The-Counter Use

(Optional Format 1-2-96)