CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody Reagent is a two-color fluorescent reagent comprised of two murine monoclonal antibodies. Each antibody is labeled with a different color fluorochrome. The reagent allows simultaneous identification and enumeration of total CD3+ and CD3-/CD56+ lymphocytes in whole blood by flow cytometry. An isotypic control, CYTO-STAT®/COULTER CLONE® MslgG1-RD1/MsIgG1-FITC, is used to monitor nonspecific staining.

CYTO-STAT®COULTER CLONE® MsIgG1-RD1/MsJgG1-PC5 Isotypic Control is a two-color fluorescent reagent comprised of two murine monoclonal antibody is labeled with a different color fluorochrome. This product is intended for use as a quality control reagent to monitor the levels of nonspecific antibody binding in cell surface staining procedures which use CYTO-STAT® or CYTO-STAT®/COULTER CLONE® Monoclonal Antibody Reagents comprised of two MsIgGl subclass monoclonal antibodies conjugated to RD1 and FITC.

CD3+ lymphocyte percentages and absolute counts may be used as aids to evaluate immune competency underlying known or unknown disease states and to monitor lymphocyte levels following organ transplantation.

To illustrate, identification of abnormal levels of CD3+ lymphocytes may aid in the diagnosis and/or prognosis of unidentified disease conditions in patients with low white blood cell counts. Measurement of CD3+ lymphocytes, in conjunction with CD4+ (inducer) and CD8+ (suppressor/cytotoxic) T lymphocytes and corresponding T4T8 ratios, may aid in the diagnosis and/or prognosis of immunodeficiency disease such as infection with human immunodeficiency virus (HV), the etiologic agent of acquired immunodeficiency syndrome (AIDS). Altered percentages of CD3+ lymphocytes recorded following organ (for example, kidney, heart, liver, lung) transplantation suggests T lymphocyte quantitation may be useful as an aid in monitoring these cellular populations.

NK (Natural Killer) lymphocyte populations have been functionally defined as a lymphocyte population capable of mediating non-MHC restricted cytoxicity against targets such as certain tumor and virus-infected cells.

As part of a Two Color Lymphocyte Immunophenotyping Panel which includes the B lymphocyte reagent, CYTO-STAT®/COULTER® CLONE® CD3(IgG1)-FITC/B4-RD1, CYTO-STAT® CD3-FITC/CD56-RD1 provides the ability to comprehensively identify and enumerate an individual's major lymphocyte subsets: T, B and NK. The reagent also finctions as a quality control check for a specimen in terms of total lymphocyte percentage and CD3+ lymphocyte measurements across the panel.

Device Description

CYTO-STAT® CD3-FITC/CD56-RD1 is a two-color fluorescent reagent comprised of two murine monoclonal antibodies. Each antibody is labeled with a different color fluorochrome. The reagent allows simultaneous identification and enumeration of total CD3+ and CD3-/CD56+ lymphocytes in whole blood by flow cytometry. An isotypic control, CYTO-STAT®/COULTER® CLONE® MsIgG1-RD1/MsIgG1-FITC, is used to monitor nonspecific staining.

CYTO-STAT®/COULTER® CLONE® MsIgG1-RD1/MsIgG1-FITC is a two-color fluorescent reagent comprised of two murine monoclonal antibodies. Each antibody is labeled with a different color fluorochrome. This product is intended for use as a quality control reagent to monitor the levels of nonspecific antibody binding in cell surface staining procedures which use CYTO-STAT® OR CYTO-STAT®/COULTER® CLONE® Monoclonal Antibody Reagents comprised of two monoclonal antibodies of the MsIgG1 subclass conjugated to RD1 and FITC.

AI/ML Overview

Acceptance Criteria and Study for CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody Reagent

This response describes the acceptance criteria and the study that proves the device meets those criteria, based on the provided text.

1. Table of Acceptance Criteria and Reported Device Performance

Acceptance Criteria	Reported Device Performance
Accuracy: Identify and enumerate appropriate numbers of targeted lymphocytes (CD3+, CD3-/CD56+, and CD19+) in whole blood specimens.	Achieved: "Results analyzed in terms of minimums, maximums, means ± 1 SD, confidence intervals, regression analyses and analyses of variance demonstrated that CD3/CD56 and CD3/B4 identify and enumerate appropriate numbers of the targeted lymphocytes in whole blood specimens." The device also provides "mature T (CD3+) lymphocyte values comparable to those of CD3/B4" and "appropriate values for NK (CD3-CD56+) and B (CD19+) lymphocytes."
Linearity: Demonstrate linearity of the assay for CD3+ and CD3-/CD56+ lymphocyte concentrations across a defined range.	Achieved: "Results analyzed in terms of regression and correlation analyses for recovered vs. expected absolute counts demonstrated Linearity of the assay."
Precision (Within Run / Intralaboratory): Demonstrate within-day precision for CD3+ and CD3-/CD56+ lymphocyte concentrations.	Achieved: "Results analyzed in terms of mean ± 1 SD and CV demonstrated Within Day (Intralaboratory) Precision of the assay."
Precision (Interlaboratory): Demonstrate interlaboratory precision for CD3+ and CD3-/CD56+ lymphocyte concentrations across different laboratories.	Achieved: "Results analyzed in terms of mean ± 1 SD and CV demonstrated Interlaboratory Precision of the assay."

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

Accuracy Study:
- Sample Size: Not explicitly stated as a number, but "Normal and abnormal whole blood specimens were collected from geographically diverse populations of males and females unselected as to race and ranging in age from 19 to 84 years."
- Data Provenance: Prospective collection from "geographically diverse populations" (country of origin not specifically mentioned, but the manufacturer is based in Miami, Florida, USA, and has international affiliates, suggesting a potential for multi-country data, though not confirmed). The data appears to be prospective as specimens were "collected" and "processed as lysed preparations and assayed in parallel."
Linearity Study:
- Sample Size: A "concentrated COULTER™ CYTO-TROL Cells sample serially diluted to achieve a range of CD3+ and CD3-/CD56+ lymphocyte concentrations." The number of dilutions or individual measurements is not specified beyond "Three replicate measurements."
- Data Provenance: Not explicitly stated, but likely laboratory-generated data using a control sample.
Precision (Within Run) Study:
- Sample Size: "Ten replicate measurements" for each of CD3+ and CD3-/CD56+ lymphocyte concentrations. The levels were obtained by "selective screening of normal whole blood specimens."
- Data Provenance: Not explicitly stated, but likely laboratory-generated data.
Precision (Interlaboratory) Study:
- Sample Size: "Ten replicate measurements" on "a single normal whole blood specimen" across "different laboratories."
- Data Provenance: Not explicitly stated, but data from multiple laboratories testing the same specimen.

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

The document does not mention the use of experts to establish ground truth for the test set. The ground truth appears to be based on comparative performance with a legally marketed predicate device (CYTO-STAT®/COULTER CLONE® CD3(IgG1)-FITC/B4-RD1) and established methods (flow cytometry and white blood cell counts/5-part differentials).

4. Adjudication Method for the Test Set

No adjudication method is described or implied in the provided text. The evaluation seems to be based on statistical analyses of the obtained measurements against those from the predicate device and expected biological ranges.

5. 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

No MRMC comparative effectiveness study was mentioned. This device is a reagent used in flow cytometry, not an AI-assisted diagnostic tool that would involve human "readers" or AI for image interpretation.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was done

This is not applicable as the device is a reagent for flow cytometry, not an algorithm. The "performance" described is of the reagent's ability to accurately identify and enumerate cell populations when used with a flow cytometer.

7. The Type of Ground Truth Used

The ground truth for the assessment of the new device (CD3/CD56) was established by comparison to:

A legally marketed predicate device: "Specimens were assayed with CD3/B4 for comparison purposes." The predicate device is identified as CYTO-STAT®/COULTER CLONE® CD3(IgG1)-FITC/B4-RD1.
Established clinical methods: "White blood cell counts and 5-part differentials were obtained for all specimens."
Biological expectations: The goal was to identify and enumerate "appropriate numbers" of targeted lymphocytes.

8. The Sample Size for the Training Set

The document does not describe a "training set" in the context of machine learning or AI. This device is a reagent, and its development would typically involve optimization and validation rather than training in the AI sense.

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

Not applicable, as there is no mention of a training set for an algorithm in this context. The development process would have involved establishing the optimal antibody concentrations and fluorochrome conjugates for accurate and specific binding.

Summary

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SEP 2 4 1998

Image /page/0/Picture/1 description: The image shows the word "COULTER" in bold, block letters. Above the word is a logo consisting of two overlapping circles with a series of right-pointing triangles in between them. Above the logo is the text "K982172" in a handwritten style. The image appears to be a logo or trademark for the Coulter brand.

COULTER CORPORATIONP.O. BOX 169015Miami, Florida 33116-9015 USA	Date:	June 17, 1998
Customer Service: (800) 526-7694Product Information: (800) 526-6932(800) 327-6531 (305) 327-6531www.coulter.com	Title:	Summary of Safety and Effectiveness Information For 510(k) Premarket Notification
	Product:	CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody ReagentwithCYTO-STAT®/COULTER® CLONE® MsIgG1-RD1/MsIgG1-FITC Isotypic Control
Coulter CorporationMiami, Florida USA	Company:	Coulter Corporation11800 SW 147 AvenueMiami, FL 33196-2500
Coulter Electronics, Pty. LtdSydney, Australia
Coulter Electronica Ind. & Com. Ltda.Rio de Janeiro, Brazil	Contact:	Dr. Marion S. Gaide (M/C: 31-B06)Senior Regulatory Affairs SpecialistCorporate Regulatory Affairs
Coulter Electronics of Canada, Ltd.Burlington, Ontario, Canada
Coulter Electronics, Ltd.Luton, Bedfordshire, England	Telephone:	(305) 380-2594
Coultronics France, S.A.Margency, France	Common or Usual or Classification Name:	Lymphocyte Immunophenotyping MonoclonalAntibody Reagent with Isotypic Control
Coulter Electronics GmbHKrefeld, Germany	Product Classification:	Product Code: GKZ; C.F.R. Section: 864.5220; ClassificationPanel: Hematology and Pathology Devices; Device Class: II
Coulter Electronics (HK), Ltd.Hong Kong
Coulter K. K.Tokyo, Japan	Intended Use:
CYTO-STAT® CD3-FITC/CD56-RD1 is a two-color fluorescent reagent comprised of two murinemonoclonal antibodies. Each antibody is labeled with a different color fluorochrome. The reagentallows simultaneous identification and enumeration of total CD3+ and CD3-/CD56+ lymphocytes in
Coulter de Mexico S.A., DE C.V.Mexico City, Mexico	whole blood by flow cytometry. An isotypic control, CYTO-STAT®/COULTER® CLONE® MsIgG1-RD1/MsIgG1-FITC, is used to monitor nonspecific staining.
Coulter Electronics, Ltd.Mijdrecht, Netherlands
Coulter Electronics, Pty. Ltd.Auckland, New Zealand		CYTO-STAT®/COULTER® CLONE® MsIgG1-RD1/MsIgG1-FITC is a two-color fluorescent
Coulter Electronics Sales of P.R., Inc.San Juan, Puerto Rico	reagent comprised of two murine monoclonal antibodies. Each antibody is labeled with a different colorfluorochrome. This product is intended for use as a quality control reagent to monitor the levels of
Coulter Electronics, Ltd.Johannesburg, South Africa	nonspecific antibody binding in cell surface staining procedures which use CYTO-STAT® OR CYTO-STAT®/COULTER® CLONE® Monoclonal Antibody Reagents comprised of two monoclonal
Coulter Electronics, Ltd.Istanbul, Turkey	antibodies of the MsIgG1 subclass conjugated to RD1 and FITC.
Coulter Electronics, S.A.Miami, Florida	Substantial Equivalence:	510(k) Premarket Notification: K926124
		CYTO-STAT®/COULTER CLONE® CD3(IgG1)-FITC/B4-RD1 Monoclonal Antibody Reagent withCYTO-STAT®/COULTER CLONE® MsIgG1-RD1/MsIgG1-FITC Isotypic Control
	Product Comparison:		The CD3/CD56 and CD3/B4 systems are essentially identical with respect to features and principles ofoperation. Each liquid reagent allows simultaneous identification and enumeration of more than one T

lymphocyte population (CD3+ and CD3-/CD56+ or CD19+) in a single specimen using a single reagent. Each system also requires an isotypic control to monitor nonspecific binding. The difference between the systems is that CD3/CD56 contains the CD56 monoclonal antibody to identify and enumerate NK (Natural Killer) lymphocytes whereas CD3/B4 contains the CD19 monoclonal antibody to identify and enumerate B lymphocytes. CD3/CD56 and CD3/B4 both require a separate reagent, CYTO-STAT®/COULTER CLONE® Mo2-RD1/KC56 (T-200)-FITC, to identify a lymphocyte gate. MAb Conjugation: CD3: FITC (Fluorescein Isothiocyanate). CD56: RD1 (Phycoerythrin).

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Product Testing:

Product testing to assess the performance of CD3/CD56 is described below. Studies were designed in line with instructions for use in the product package insert and performance specifications. Specimens were assayed with CD3/B4 for comparison purposes. The results of product testing demonstrated that CD3/CD56 meets all performance specifications and provides mature T (CD3+) lymphocyte values comparable to those of CD3/B4. CD3/CD56 and CD3/B4 also provide appropriate values for NK (CD3-CD56+) and B (CD19+) lymphocytes.

1. Accuracy:
  Normal and abnormal whole blood specimens were collected from geographically diverse populations of males and females unselected as to race and ranging in age from 19 to 84 years. Specimens were divided, processed as lysed preparations and assayed in parallel with CD3/CD56 and CD3/B4. CD3+, CD3-/CD56+ and CD19+ percentages expressed in terms of the total lymphocyte count and absolute counts (cells/uL) were determined with COULTER® EPICS® XL-MCL™ flow cytometers gated on lymphocytes. White blood cell counts and 5-part differentials were obtained for all specimens.

Results analyzed in terms of minimums, maximums, means ± 1 SD, confidence intervals, regression analyses and analyses of variance demonstrated that CD3/CD56 and CD3/B4 identify and enumerate appropriate numbers of the targeted lymphocytes in whole blood specimens.

1. Linearity:
  Three replicate measurements were made on a concentrated COULTER™ CYTO-TROL Cells sample serially diluted to achieve a range of CD3+ and CD3-/CD56+ lymphocvte concentrations. Samples were assayed with CD3/CD56 and analyzed on a COULTER® EPICS® XL-MCL™ flow cytometer gated on lymphocytes. Values were expressed in terms of absolute count (cells/uL).

Results analyzed in terms of regression and correlation analyses for recovered vs. expected absolute counts demonstrated Linearity of the assay.

Precision: Within Run (Intralaboratory): 3.
Ten replicate measurements were made for each of CD3+ and CD3-/CD3+ and CD3-/CD56+ lymphocyte concentrations on the same day using a COULTER® EPICS® XL-MCL™ flow cytometer gated on lymphocytes. Levels were obtained by selective screening of normal whole blood specimens and assayed with CD3/CD56. Values were expressed in terms of percentage of the total lymphocyte count.

Results analyzed in terms of mean ± 1 SD and CV demonstrated Within Day (Intralaboratory) Precision of the assay.

1. Precision (Interlaboratory):
  Ten replicate measurements on were made on the same day using different laboratories and COULTER® EPICS® XL-MCL™ flow cytometers. All measurements were made on a single normal whole blood specimen divided and assayed with CD3/CD56. Values were expressed in terms of percentage of the total lymphocyte count.

Results analyzed in terms of mean ± 1 SD and CV demonstrated Interlaboratory Precision of the assay.

Marion S. Gaide, Ph.D.

Marion S. Gaide Ph.D. Senior Regulatory Affairs Specialist Corporate Regulatory Affairs

June 17, 1998
Date

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SEP 2 4 1998

Marion S. Gaide, Ph.D. Senior Regulatory Affairs Specialist Coulter Corporation 11800 SW 147 Avenue Miami, Florida 33196-2500

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Re : K982172/S1

Trade Name: CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody Reagent with CYTO-STAT®/COULTER® CLONE® MsIqG1-RD1/MsIgG1-FITC Isotypic Control Regulatory Class: II Product Code: GKZ Dated: August 19, 1998 Received: Auqust 21, 1998

Dear Dr. Gaide: . . .....

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 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 Requlations, 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. 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 i 7 : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : for devices under the Electronic Product Radiation Control provisions, ... ...... or other Federal laws or regulations.

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This letter will allow you to beqin 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 requlation 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/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Putman

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

510(k) Number (if known): Not Yet Assigned

CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody Reagent Device Name: with CYTO-STAT®/COULTER® CLONE® MsIgG1-RD1/MsIgG1-FITC Isotypic Control

Indications For Use:

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

Prescription Use(Per 21 CFR 801.109)
	Over-The-Counter Use

Concurrence of CDRH, Office of Device Evaluation (ODE)

(Division Sign-Off)
Division of Clinical Laboratory Devices
510(k) Number	K982172

510i4us2

Regulation Number and Section

§ 864.5220 Automated differential cell counter.

(a)
Identification. An automated differential cell counter is a device used to identify one or more of the formed elements of the blood. The device may also have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood, bone marrow, or other body fluids. These devices may combine an electronic particle counting method, optical method, or a flow cytometric method utilizing monoclonal CD (cluster designation) markers. The device includes accessory CD markers.(b)
Classification. Class II (special controls). The special control for this device is the FDA document entitled “Class II Special Controls Guidance Document: Premarket Notifications for Automated Differential Cell Counters for Immature or Abnormal Blood Cells; Final Guidance for Industry and FDA.”