CYTO-STAT TRICHROME CD45-FITC/CD56-RD1/CD3-PC5 MONOCLONAL ANTIBODY REAGENT WITH CYTO-STAT TRICHROME CD45-FITC/MSIGG1-RD1 by COULTER CORP.

CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 Monoclonal Antibody Reagent is a three-color fluorescent reagent comprised of three murine monoclonal antibody is labeled with a different color fluorochrome. The reagent allows simultaneous identification and enumeration of total CD3+CD56+ lymphocytes in whole blood by flow cytometry. An isotypic control, CYTO-STAT® triCHROME™ CD45-FITC/MsIgG1-PC5, is used to monitor nonspecific staining.

CYTO-STAT® triCHROME™ CD45-FITCMsIgG1-RD1/MsIgG1-PC5 Isotypic Control is a three-color fluoresent reagent comprised of three 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® triCHROME™ Monoclonal Antibody Reagents comprised of CD45-FITC and two MsIgG1 subclass monoclonal antibodies conjugated to RD1 and PC5.

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

To illustrate, identification of abnormal levels of CD3+ lymphocytes may aid in the diagnosis and/or progrosis of unidentified disease conditions in patients with low white blood cell counts. Measurement of CD3+ lymphocytes, in conjunction with CD4+ (induce) and CD8+ (suppressor/cytoxic) T lymphocytes and corresponding T478 ratios, may aid in the diagnois and/or prognosis of immunodeficiency disease such as infection with human immunodeficiency virus (HIV), 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 Three Color Lymphocyte Immunophenotyping Panel which includes the B lymphocyte reagent, CYTO-STAT® triCHROME™ CD45-FITC/CD19-RD1/CD3-PC5, CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 provides the ability to comprehensively identify and enumerate an individual's major lymphocyte subsets: T, B and NK. The reagent also functions 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® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 is a three-color fluorescent reagent comprised of three murine monoclonal antibodies. Each antibody is labeled with a different color fluorochrome. The reagent allows simultaneous identification and enumeration of total CD3+ and CD3-.

CYTO-STAT® triCHROME™ CD45-FITC/MsIgG1-RD1/MsIgG1-PC5 Isotypic Control is a three-color fluorescent reagent comprised of three 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® triCHROME™ Monoclonal Antibody Reagents comprised of CD45-FITC and two monoclonal antibodies conjugated to RD1 and PC5.

AI/ML Overview

This document describes the safety and effectiveness information for the NOV COULTER CORPORATION's CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 Monoclonal Antibody Reagent with its corresponding Isotypic Control. The information provided focuses on the product's performance specifications and how it meets them.

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are implied by the "performance specifications" that the device is stated to meet. The study's results demonstrated that the device successfully met these specifications.

Acceptance Criteria Category	Specific Criteria (Implied)	Reported Device Performance
Accuracy	CD45/CD56/CD3 should identify and enumerate essentially identical numbers of targeted lymphocytes in whole blood specimens compared to the predicate device (CD3/CD56).	"Results analyzed in terms of minimums, maximums, means ± 1 SD, confidence intervals, regression analyses and analyses of variance demonstrated that CD45/CD3 and CD3/CD56 identify and enumerate essentially identical numbers of the targeted lymphocytes in whole blood specimens." This was for both CD3+ and CD3-/CD56+ percentages and absolute counts.
Linearity	The assay should demonstrate linearity across a range of lymphocyte concentrations for CD3+ and CD3-/CD56+ populations.	"Results analyzed in terms of regression and correlation analyses for recovered vs. expected absolute counts demonstrated Linearity of the assay."
Precision (Within-Run)	The assay should demonstrate acceptable precision (mean ± 1 SD and CV) for replicate measurements of CD3+ and CD3-/CD56+ lymphocyte concentrations within a single lab and run.	"Results analyzed in terms of mean ± 1 SD and CV demonstrated Within Day (Intralaboratory) Precision of the assay."
Precision (Interlaboratory)	The assay should demonstrate acceptable precision (mean ± 1 SD and CV) for replicate measurements of CD3+ and CD3-/CD56+ lymphocyte concentrations across different laboratories.	"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

Sample Size: The document states that "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." However, the exact number of specimens/samples used for the accuracy, linearity, and precision studies is not explicitly stated. For the within-run precision, "Ten replicate measurements were made for each of three levels of CD3+ and CD3-/CD56+ lymphocyte concentrations." For interlaboratory precision, "Ten replicate measurements on were made on the same day."
Data Provenance: The specimens were collected from "geographically diverse populations." The document does not specify particular countries but implies a broad collection. The study appears to be prospective in nature, as specimens were "collected" and then "processed as lysed preparations and assayed in parallel" for the purpose of this product testing.

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

This type of device (monoclonal antibody reagent for flow cytometry) relies on established cellular markers and flow cytometry principles rather than subjective expert interpretation for ground truth. Therefore, there's no mention of "experts" in the traditional sense of adjudicating images or clinical cases. The "ground truth" is inherently tied to the established scientific understanding of CD antigens and their detection via flow cytometry. The reference device, CYTO-STAT® CD3/CD56, serves as the comparison for establishing equivalence, implying its performance is considered the de facto "truth" against which the new device is measured.

4. Adjudication Method for the Test Set

Not applicable for this type of in vitro diagnostic device study. The "ground truth" is determined by the measurements obtained from the predicate device and the consistent application of flow cytometry principles. The performance of the new device is directly compared to these measurements.

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

Not applicable. This is an in vitro diagnostic reagent, not an AI-assisted diagnostic tool that involves human readers interpreting results.

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

Not applicable. This is a reagent for flow cytometry, not an algorithm. The "performance" is the ability of the reagent to accurately label cells for subsequent analysis by a flow cytometer, which produces quantitative data. Human operators are involved in specimen preparation, running the flow cytometer, and interpreting the raw flow cytometry data (e.g., gating lymphocytes), but the reagent itself is not a standalone algorithm.

7. The Type of Ground Truth Used

The ground truth used is comparison to a legally marketed predicate device (CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody Reagent with CYTO-STAT®/COULTER CLONE® MsIgG1-RD1/MsIgG1-FITC Isotypic Control). The performance of the new device (CD45/CD56/CD3) was compared to this predicate device (CD3/CD56) using flow cytometry measurements, which are considered quantitative and objective measures for cell populations.

8. The Sample Size for the Training Set

Not applicable. This is a reagent, not an AI model that requires a training set. The development of the reagent involves chemical and biological formulation, not data training.

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

Not applicable, as there is no training set for a reagent product.

Summary

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NOV COULTER CORPORATION P.O. BOX 169015 Date: November 4, 1998 Miami Florida 33116-9015 USA Customer Service: (800) 526-769 Summary of Safety and Effectiveness Information For 510(k) Premarket Notification Title: Product Information: (800) 526-6932 (800) 327-6531 (305) 357-653 www.coulicr.com Product: CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 Monoclonal Antibody Reagent with CYTO-STAT® triCHROME™ CD45-FITC/MsIgG1-RDJ/MsIgG1-PC5 Isotypic Control Coulter Corporation Company; 10.17 11 11 11 11 11 11 11 11 11800 SW 147 Avenue Miami, FL 33196-2500 ... ... . . Contact: Dr. Marion S. Gaide (M/C: 31-B06) Senior Regulatory Affairs Specialist 1. . . . . ( ' ' . . . Sun 10 11 11 Corporate Regulatory Affairs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Yollars for note to be and 12. I mades! (, addrivers , trans , Telephone: (305) 380-2594 Mary College Comments of : . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lymphocyte Immunophenotyping Monoclonal Antibody Reagent with Isotypic Control Kray at to marker Classified on the 2014 Product Classification: Product Code: GKZ; C.F.R. Section: 864.5220; Classification Hematology and Pathology Devices; Device Class: II Panel: いた。 Produkt K
100000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000 Come of Moscore Matther CYTO STATE Conceller CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 is a three-color fluorescont reagent · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · time in thorochrome. The reagent allows simultancous identification and enumeration of total CD3+ and CD3-. ... .. .. .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . No do C V a la triCHROMETM CD45-FITC/MsIgG1-PCS, is used to monitor nonspecific staining. range of the count of the triCHROME CD45-FITCMsIgGT+RD1/MsgG1-PCS is a threesed in filloresear color in we lighted of three inutine monoclonal antibodies. Each antibody is labeled with a different store of the color fluorochrome. This product is intended for use as a quality control reagent to monitor the levels of Course In nonspecific antibody binding in cell surface staining procedures which use CYTO-STAT® a monoloriCHROME™ Monoclonal Antibody Reagents comprised of CD45-FITC and two monoclonal Colletter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Caracta, Vanchestia Substantial Equivalence: 510(k) Premarket Notification: K982172 CYTO-STAT® CD3-FITC/CD56-RD1 Monoclonal Antibody Reagent with CYTO-STAT®/COULTER CLONE® MsIgG1-RD1/MsIgG1-FITC Isotypic Control Product Comparison: The CD45/CD56/CD3 and CD3/CD56 systems are essentially identical with respect to features and principles of operation. Each liquid reagent allows simultaneous identification and enumeration of more than one lymphocyte population (CD3+ and CD3-/CD56+) in a single specimen using a single reagent. Bach system also requires an isotypic control to monitor nonspecific binding. The difference between the systems is that CD45/CD36/CD3 contains CD45 to identify a lymphocyte gate for making CD3+ and CD3-/CD56+ measurcments. CD3/CD56 rcquires a scparate reagent, CYTO-STAT®/COULTER CLONE® Mo2-RD1/KCS6 (T-200)-FITC, for this purpose. MAb Conjugation: CD45: FITC

(Fluorescein Isothiocyanate). CD56: RD1 (Phycoerythrin). CD3: PCS (Phycoerythrin-Cy5),

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Product testing to assess the performance of CDAS/CD3 is described below. Studies were Product Testing: designed in line with instructions for use in the product package insert and performance specifications, Specimens were assayed with CD3/CD56 for comparison purposes. The results of product testing demonstrated that CD45/CD56/CD3 meets all performance specifications and provides mature T (CD3+) and NK (CD3-/CD56+) lymphocyte values comparable to those of CD3/CD56.
- l . 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 CD45/CD56/CD3 and CD3/CD56. CD3+ and CD3-/CD56+ percentages expressed in terms of the total lymphocyte count and absolute counts (cells/uL) were determined with COULTER® EPICS® XL-MCL ™ flow cytomerers 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 CD45/CD3 and CD3/CD56 identify and enumerate essentially identical numbers of the targeted lymphocytes in whole blood specimens.

1. Linearity:
  Three replicate measurements were made on a concentrated COULTER™ CYTO-TROL Cclls sample serially diluted to achieve a range of CD3+ and CD3-/CD56+ lymphocyte concentrations, Samples were assayed with CD45/CD56/CD3 and analyzed on a COULTER® EPICS® XL-MCL™ flow cytometer gated on lymphocytes. Values were expressed in terms of absolute count (cclls/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 three levels of 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 CD45/CD56/CD3. 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):
  Ton replicate measurements on were made on the same day using different laboratories and COULTER® EPICS® XL-MCL™ flow cytomcters. All measurements were made on a single normal whole blood specimen divided and assayed with CD45/CD56/CD3, 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. Gaido, Ph.D.

Senior Regulatory Affairs Specialist Corporate Regulatory Affairs

November 4, 1998
Date

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Received: October 1, 1998

Image /page/2/Picture/2 description: The image shows the logo for the Department of Health & Human Services USA. The logo features a stylized eagle with three stripes extending from its back, symbolizing the department's commitment to health and well-being. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" is arranged in a circular pattern around the eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Marion S. Gaide, Ph.D. Senior Regulatory Affairs Specialist Coulter Corporation P.O. Box 169015 NOV રે 1998 Miami, Florida 33116-9015 Re : к982166/S1 Trade Name: CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 Monoclonal Antibody Reagent with CYTO-STAT® triCHROME™ CD45-FITC/MsIqG1-RD1/MsIgG1-PC5 Isotypic Control Regulatory Class: II Product Code: GKZ Dated: September 29, 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 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 A substantially equivalent determination assumes compliance to 895. with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General requlation (21 CFR Part 820) and that, throuqh periodic QS inspections, the Food and Druq 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.

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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/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Butman

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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Page 1 of 1

INDICATIONS FOR USE

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

CYTO-STAT® triCHROME™ CD45-FITC/CD56-RD1/CD3-PC5 Monoclonal Antibody Reagent Device Name: with CYTO-STAT® triCHROME™ CD45-FITC/MsIgG1-RD1/MsIgG1-PC5 Isotypic Control

Indications For Use:

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

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

(Division Sign-Off)Division Sian-Om Division of Clinical Laboratory Devices 510/k) Nimber

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.”