K Number

Device Name

CYTO-STAT TRICHROME CD8-FITC/CD4-RDI/CD3-PC5 MONOCLONAL ANTIBODY REAGENT WITH CYTO-STAT TRICHROME MSIGGI-FITC/MSIGGI-RD1

Manufacturer

COULTER CORP.

Date Cleared

1996-12-23

(69 days)

Product Code

GKZ

Regulation Number

864.5220

Intended Use

CYTO-STAT® triCHROME™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody Reagent 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+, total CD4+, total CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow cytometry. An isotypic control, CYTO-STAT® triCHROME™ MsIgG1-FITC/MsIgG1-RD1/MsIgG1-PC5, is used to monitor nonspecific staining. CYTO-STAT® triCHROME™ MsIgG1-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™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody Reagent to identify and enumerate total CD3+, total CD4+, total CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow cytometry.

Device Description

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

K950742

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device is a reagent for flow cytometry, which is a laboratory technique. The description focuses on the antibodies and their use in identifying and enumerating cell populations, not on any computational analysis involving AI/ML. The performance studies describe standard statistical analyses of accuracy and precision, not AI/ML model performance metrics.

Therapeutic

No.
The device is a diagnostic reagent used for identification and enumeration of lymphocytes, not for therapy or treatment.

Diagnostic

Yes

This device is used to identify and enumerate specific lymphocyte populations in whole blood by flow cytometry, including total CD3+, total CD4+, total CD8+, dual CD3+/CD4+, and dual CD3+/CD8+ lymphocytes. This information can be used to assess the immune status of individuals, which is a diagnostic purpose. The Intended Use section also explicitly states its use in distinguishing normal and abnormal blood samples, further supporting its diagnostic nature.

SaMD (Software as a Medical Device)

The device description clearly states it is a "three-color fluorescent reagent comprised of three murine monoclonal antibodies," which are physical components, not software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The intended use explicitly states that the reagent is used for the "simultaneous identification and enumeration of total CD3+, total CD4+, total CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow cytometry." This is a diagnostic purpose, providing information about a patient's immune status.
Sample Type: The device is used with "whole blood," which is a biological specimen taken from the human body.
Method: The method used is "flow cytometry," which is a common laboratory technique for analyzing cells.
Control: The inclusion of an "isotypic control" further indicates its use in a diagnostic setting to ensure the validity of the results.
Performance Studies: The document describes performance studies (Accuracy, Linearity, Precision) which are typical for demonstrating the reliability and validity of an IVD.
Predicate Device: The mention of a "Predicate Device" with a K number (K950742) strongly suggests that this device is being submitted for regulatory clearance as an IVD, as predicate devices are used for comparison in the 510(k) submission process for IVDs in the United States.

All these factors align with the definition and characteristics of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

CYTO-STAT® triCHROME™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody Reagent is a three-color fluorescent reagent comprised of three murine monoclona antibodies. Each antibody is labeled with a different color fluorochrome. The reagen CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow CYTO-STAT® triCHROME™ isotypic control. MsIgG1cytometry. An FITC/MsIgG1-RD1/MsIgG1-PC5, is used to monitor nonspecific staining. CYTO-STAT® triCHROME™ MsIgG1-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 allows simultaneous identification and enumeration of total CD3+, total CD4+, total CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow cytometry.

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™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody Reagent to identify and enumerate total CD3+, total CD4+, total CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow cytometry.

Product codes (comma separated list FDA assigned to the subject device)

GKZ

Device Description

CYTO-STAT® triCHROME™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody Reagent is a three-color fluorescent reagent comprised of three murine monoclonal antibodies. Each antibody is labeled with a different color fluorochrome. CYTO-STAT® triCHROME™ MsIgG1-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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Whole blood

Indicated Patient Age Range

18 to 85 years

Intended User / Care Setting

Not Found

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

Normal and abnormal (e.g., Human Immunodeficiency Virus, organ transplant, autoimmune disease, low white blood cell count) whole blood specimens were collected from geographically diverse populations of males and females unselected as to race and ranging in age from 18 to 85 years. Specimens were divided, processed as lysed preparations and assayed in parallel with CD8/CD4/CD3 and CD3/T4/T8. The CD3+, CD4+, CD3+/CD4+ and CD3+/CD8+ percentages expressed in terms of the total lymphocyte count and absolute counts (cells/pL) were determined with a COULTER® EPICS® XL-MCL flow cytometer gated on lymphocytes. White blood cell counts and 5-part differentials were obtained for all specimens using the COULTER® STKS. Absolute counts were determined using both the Standard (Indirect) Method, and the Flow-Count (Direct) Method with Flow-Count™ Fluorospheres. All values were corrected for lymphocyte purity (Lymphocyte Gate Limits: lymphocyte recovery ≥ 90%; lymphocyte purity ≥ 85%).

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

Accuracy:
Normal and abnormal (e.g., Human Immunodeficiency Virus, organ transplant, autoimmune disease, low white blood cell count) whole blood specimens were collected from geographically diverse populations of males and females unselected as to race and ranging in age from 18 to 85 years. Specimens were divided, processed as lysed preparations and assayed in parallel with CD8/CD4/CD3 and CD3/T4/T8. The CD3+, CD4+, CD3+/CD4+ and CD3+/CD8+ percentages expressed in terms of the total lymphocyte count and absolute counts (cells/pL) were determined with a COULTER® EPICS® XL-MCL flow cytometer gated on lymphocytes. White blood cell counts and 5-part differentials were obtained for all specimens using the COULTER® STKS. Absolute counts were determined using both the Standard (Indirect) Method, and the Flow-Count (Direct) Method with Flow-Count™ Fluorospheres. All values were corrected for lymphocyte purity (Lymphocyte Gate Limits: lymphocyte recovery ≥ 90%; lymphocyte purity ≥ 85%).
Results analyzed in terms of minimums, means ± 1 SD, confidence intervals.with 95% limits, regression and correlation analyses of variance, demonstrated that CD8/CD4/CD3 and CD3/T4/T8 identify and enumerate essentially identical numbers of the targeted lymphocytes in whole blood specimens.

Linearity:
Three replicate measurements were made on a concentrated COULTER™ CYTO-TROL™ Control Cells sample serially diluted to achieve a range of CD3+, CD4+ (CD3+/CD4+) and CD8+ (CD3+/CD8+) lymphocyte concentrations. Samples were assayed with CD&/CD4/CD3 and analyzed on a COULTER® EPICS® XL-MCL flow cytometer gated on lymphocytes. Values were expressed in terms of absolute counts (cells/uL).
Results analyzed in terms of regression and correlation analyses for recovered versus expected absolute counts demonstrated linearity of the assay.

Within Run (Intralaboratory) Precision:
Ten replicate measurements were made for each of three levels of CD3+, CD4+ (CD3+/CD4+) and CD8+ (CD3+/CD8+) lymphocyte concentrations using a COULTER® EPICS® XL-MCL flow cytometer gated on lymphocytes. Levels were obtained by selective depletions of a normal whole blood specimen and assaved with CD8/CD4/CD3. Values were expressed in terms of % of the total lymphocyte count.
Results analyzed in terms of mean ± 1 SD and CV demonstrated Within Run (Intralaboratory) Precision of the assay.

Interlaboratory Precision:
Ten replicate measurements 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 CD8/CD4/CD3. Values were expressed in terms of % of the total lymphocvte count.
Results analyzed in terms of mean ± 1 SD and CV demonstrated Interlaboratory Precision of the assav.

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

Not Found

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

K950742

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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.”

Summary

K964618

COULTER CORPORATION P.O. BOX 169015 Mlami, Florida 33116-9015 USA

Date: October 11, 1996

Summary of Safety and Effectiveness Information for Title: "K950742 510(k) Additional Information"

Prod Coulter Corporation Miami, Florida USA

Coulter Leasing Corporation Miami, Fiorida USA

Coulter Electronics, Pty. Lid. Sydney, Australia

Coulter Electronics Ind. & Com., Ltda Rio de Janeiro, Brazil

Coulter Electronics of Canada, Ltd. Burlington, Ontario. Canada

Coulter Electronics, i.td Luton Bedfordshire, England

Coultronics France, S.A. Margency, France

Coulter Electronics GinbH Krefeld, Germany

Coulter Electronics (HK). Ltd Hono Kong

Coulter K. K. Tokyo, Japan

Coulter de Mexico S.A., DE C.V. Mexico City. Mexico

Coulter Electronics. Ltd Mijdrecht. Netherlands

Coulter Electronics, Piv, Ltd. Auckland, New Zealarid

Coulter Electronics Sales of P.R., Inc. San Juan, Puerto Rico

Coulter Electronics. Lid Johannesburg, South Africa

Coulter Electronics 11ct Istanbur Turkey

Coulter Electronic: 0 6 Carabas, Venerial

| Product: | CYTO-STAT ® triCHROME™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody Reagent
with CYTO-STAT® triCHROME™ MsIgG1-FITC/MsIgG1-RD1/MsIgG1-PC5 Isotypic Control | |
|------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|
| Company: | Coulter Corporation
11800 SW 147 Avenue
Miami, FL 33196-2500 | |
| Contact: | Dr. Marion S. Gaide (M/C: 31-B06)
Senior Regulatory Affairs Specialist
Corporate Regulatory Affairs | |
| Telephone: | 305-380-2594 | |
| Common or Usual or Classification Name: | Lymphocyte Immunophenotyping Monoclonal
Antibody Reagent and Isotypic Control | |
| Product Classification: | Product Code: GKZ; C.F.R. Section: 864.5220
Classification Panel: Hematology and Pathology Devices; Device Class: II | |
| Intended Use: | CYTO-STAT® triCHROME™ CD8-FITC/CD4-RD1/CD3-PC5 Monoclonal Antibody
Reagent is a three-color fluorescent reagent comprised of three murine monoclona
antibodies. Each antibody is labeled with a different color fluorochrome. The reagen | |

CD8+, dual CD3+/CD4+ and dual CD3+/CD8+ lymphocytes in whole blood by flow CYTO-STAT® triCHROME™ isotypic control. MsIgG1cytometry. An FITC/MsIgG1-RD1/MsIgG1-PC5, is used to monitor nonspecific staining. CYTO-STAT® triCHROME™ MsIgG1-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

allows simultaneous identification and enumeration of total CD3+, total CD4+, total

Substantial Equivalence: "K950742 510(k) Premarket Notification"

CYTO-STAT®/COULTER CLONE® CD3-ECD/T4-RD1/T8-FITC Monoclonal Antibody Reagent with

CYTO-STAT®/COULTER CLONE® MsIgG2b-ECD/MsIgG1-RD1/MsIgG1-FFTC Isotypic Control

Product Differences: CD8/CD4/CD3 with MsIgG1/MsIgG1/MsIgG1 and CD3/T4/T8 with MsIgG2b/MsIgG1/MsIgG1 are essentially identical with respect to features and principles of operation. Both the revised and original product systems use the same, well-established, state-of-the-art technologies of immunophenotyping with monoclonal antibodies and flow cytometry to measure cellular components in whole blood via immunofluorescence analysis. Further, the intended use of each system is the same. The liguid monoclonal antibody reagents allow simultaneous identification and enumeration of more than one lymphocyte population (total CD3+, total CD4+, total CD8+, dual CD3+/CD4+ and dual CD3+/CD8+) in a single specimen using a single reagent. Each system also uses separate reagents to 1) identify a lymphocyte gate (i.e., CyTo-STAT®/COULTER CLONE® Mo2-RD1/KC56 (T-200)-FITC); and 2) monitor nonspecific binding (i.e., isotypic control).

The few differences (italicized) between the reagent and isotypic control formulations are as follows:

| 1. CD3 Clone: | CD8/CD4/CD3:
CD3/T4/T8: | UCHTI
HIT3a |
|------------------------------|-------------------------------------------------|-------------------------------------------------------------|
| 2. CD3 Isotype: | CD8/CD4/CD3:
CD3/T4/T8: | MslgG1
MsIgG2a |
| 3. Isotypic Control Clone: | MsIgG1/MsIgG1/MsIgG1:
MsIgG2b/MsIgG1/MsIgG1: | ----------; MsIgG1=2T8-2F5
MsIgG2b=MPC11; MsIgG1=2T8-2F5 |
| 4. Isotypic Control Isotype: | MsIgG1/MsIgG1/MsIgG1:
MsIgG2b/MsIgG1/MsIgG1: | ----------; MsIgG1
MslgGb; MsIgG1 |
| 5. CD3 Fluorochrome: | CD8/CD4/CD3:
CD3/T4/T8: | PC5 (Phycoerythrin-Cy5)
ECD (Energy-Coupled Dye) |

Product testing to assess the performance of the CD8/CD3 product system is described below. Product Testing: Studies were designed in line with instructions for use given in the Package Inserts, Product Manuals, and performance specifications. Specimens were assayed with the CD3/T4/T8 product system for comparison purposes. The results of product testing demonstrated that CD8/CD4/CD3 met all performance specifications and provided mature T (CD3+), inducer T (CD4+; dual CD3+/CD4+) and suppressor/cytotoxic T (CD8+; dual CD3+/CD8+) T lymphocyte values comparable to those of CD3/T4/T8.
- 1. Accuracy:

Results analyzed in terms of minimums, means ± 1 SD, confidence intervals.with 95% limits, regression and correlation analyses of variance, demonstrated that CD8/CD4/CD3 and CD3/T4/T8 identify and enumerate essentially identical numbers of the targeted lymphocytes in whole blood specimens.

Linearity: 2.
Three replicate measurements were made on a concentrated COULTER™ CYTO-TROL™ Control Cells sample serially diluted to achieve a range of CD3+, CD4+ (CD3+/CD4+) and CD8+ (CD3+/CD8+) lymphocyte concentrations. Samples were assayed with CD&/CD4/CD3 and analyzed on a COULTER® EPICS® XL-MCL flow cytometer gated on lymphocytes. Values were expressed in terms of absolute counts (cells/uL).

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

1. Within Run (Intralaboratory) Precision:
  Ten replicate measurements were made for each of three levels of CD3+, CD4+ (CD3+/CD4+) and CD8+ (CD3+/CD8+) lymphocyte concentrations using a COULTER® EPICS® XL-MCL flow cytometer gated on lymphocytes. Levels were obtained by selective depletions of a normal whole blood specimen and assaved with CD8/CD4/CD3. Values were expressed in terms of % of the total lymphocyte count.

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

1. Interlaboratory Precision:
  Ten replicate measurements 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 CD8/CD4/CD3. Values were expressed in terms of % of the total lymphocvte count.

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

Marion S. Gaede

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

October 11, 1996