K Number

Device Name

TRITEST CD3 FITC/CD19 PE/CD45 PERCP REAGENT WITH TRUCOUNT ABSOLUTE COUNT TUBES

Manufacturer

BECTON DICKINSON IMMUNOCYTOMETRY SYSTEMS

Date Cleared

1997-10-22

(236 days)

Product Code

GKZ

Regulation Number

864.5220

Intended Use

For in vitro diagnostic use to identify and enumerate percentages and absolute counts of T and B lymphocytes in bload. Indications for Use - For use with any flow cytometer with specified detection ranges . - For use with erythrocyte lysed whole blood . - For use with or without an isotype control . - For in vitro diagnostic use . - To identify and enumerate percentages and absolute counts of CD3+ and CD19+ ● lymphocytes - To characterize and monitor some forms of immunodeficiency . - To characterize and monitor some forms of autoimmune diseases .

Device Description

The BDIS TriTEST CD3 fluorescein isothiocyanate (FITC)/CD19 phycoerythrin (PE)/CD45 peridinin chlorophyll protein (PeCP) reagent is a three-color, direct immunofluorescence reagent for identifying and enumerating percentages of T lymphocytes (CD3+) and B lymphocytes (CD19+) in eythrocyte-lysed whole blood (LWB). When used with TRUCOUNT Absolute Count Tubes, the product will yield absolute counts in cells/uL. The Becton Dickinson Tril EST / TRUCOUNT system for immunophenotyping consists of a flow cytometer (cither from BDIS or from another manufacturer), conjugated monoclonal reagent (TriTEST CD3 FITC/CD19 PE/CD45 PeCP) and TRUCOUNT Absolute Count Tubes. The process to obtain lymphocyte subset percentages includes: 1) obtaining a whole blood sample, 2) cell-surface antigen staining with three-color monoclonal antibody reagents, 3) erythrocyte lysis, and 4) flow cytometric acquisition and analysis of list mode data. Analysis involves computing the ratio of reagent-positive events (CD3+ and CD19+) to the CD45 positive events, and expressing the ratio as a percentage. To obtain absolute counts, the TriTEST reagent and whole blood are added directly to an Absolute Count Tube prior to lysis. The remaining process, until analysis, is identical to that for percentages. Analysis for absolute counts requires that an additional region, the bead region, be identified and the events in this region counted. The proportion of reagent positive events to be identified and are trated. The absolute count is P x (beads/pellet)/(volume of blood sample). When monoclonal antibody reagents are added to human whole blood, the fluorochromelabeled antibodies bind specifically to antigens on the surface of leucocytes, thus identifying lymphocyte populations. The patient blood sample is added to the counting bead peller and is treated with fluorochrome-labeled antibodies and the erythrocytes are lysed with FACS® Lysing Solution. The flow cytometer is set up so that cell populations for most samples occupy approximately the same region of fluorescence space. The sample is then introduced into the flow cytometer and the stained cells and beads fluoresce when excited by a laser beam. The three-color reagent permits identification of lymphocyte subsets using fluorescence gating instead of forward scatter gating. This three-color reagent allows direct gating on the CD45-positive population using a combination of fluorescence and side scatter parameters. By gating on the CD45-positive population, a maximum number of lymphocytes may be captured in the gate and non-lymphocyte contamination may be minimized.

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

K913192, K950342

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on traditional flow cytometry methods and reagent-based analysis, with no mention of AI or ML algorithms for data processing or interpretation.

Therapeutic

No
This device is for in vitro diagnostic use, intended to identify and enumerate percentages and absolute counts of T and B lymphocytes in blood to characterize and monitor some forms of immunodeficiency and autoimmune diseases. It is not used to treat or prevent a disease or condition.

Diagnostic

Yes
The "Intended Use / Indications for Use" section explicitly states, "For in vitro diagnostic use." Additionally, the device is indicated "To characterize and monitor some forms of immunodeficiency" and "To characterize and monitor some forms of autoimmune diseases," which are diagnostic purposes.

SaMD (Software as a Medical Device)

The device description clearly states it is a "reagent" and includes physical components like "conjugated monoclonal reagent" and "TRUCOUNT Absolute Count Tubes." It is used in conjunction with a flow cytometer, which is a hardware device. The process involves physical steps like staining, lysis, and flow cytometric acquisition.

IVD (In Vitro Diagnostic)

Based on the provided text, the device is an IVD (In Vitro Diagnostic).

Here's why:

Explicit Statement: The "Intended Use / Indications for Use" section explicitly states: "For in vitro diagnostic use to identify and enumerate percentages and absolute counts of T and B lymphocytes in bload." and "For in vitro diagnostic use".
Device Description: The description details a process for analyzing human whole blood samples using reagents and a flow cytometer to identify and quantify specific cell populations (T and B lymphocytes). This is a typical in vitro diagnostic procedure.
Indications for Use: The indications for use relate to characterizing and monitoring medical conditions (immunodeficiency, autoimmune diseases) based on the analysis of blood samples, which is a diagnostic application.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

For in vitro diagnostic use to identify and enumerate percentages and absolute counts of T and B lymphocytes in bload.

Indications for Use

For use with any flow cytometer with specified detection ranges .
For use with erythrocyte lysed whole blood .
For use with or without an isotype control .
For in vitro diagnostic use .
To identify and enumerate percentages and absolute counts of CD3+ and CD19+ ● lymphocytes
To characterize and monitor some forms of immunodeficiency .
To characterize and monitor some forms of autoimmune diseases .

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

GKZ

Device Description

The process to obtain lymphocyte subset percentages includes: 1) obtaining a whole blood sample, 2) cell-surface antigen staining with three-color monoclonal antibody reagents, 3) erythrocyte lysis, and 4) flow cytometric acquisition and analysis of list mode data. Analysis involves computing the ratio of reagent-positive events (CD3+ and CD19+) to the CD45 positive events, and expressing the ratio as a percentage.

To obtain absolute counts, the TriTEST reagent and whole blood are added directly to an Absolute Count Tube prior to lysis. The remaining process, until analysis, is identical to that for percentages. Analysis for absolute counts requires that an additional region, the bead region, be identified and the events in this region counted. The proportion of reagent positive events to be identified and are trated. The absolute count is P x (beads/pellet)/(volume of blood sample).

When monoclonal antibody reagents are added to human whole blood, the fluorochromelabeled antibodies bind specifically to antigens on the surface of leucocytes, thus identifying lymphocyte populations. The patient blood sample is added to the counting bead peller and is treated with fluorochrome-labeled antibodies and the erythrocytes are lysed with FACS® Lysing Solution. The flow cytometer is set up so that cell populations for most samples occupy approximately the same region of fluorescence space. The sample is then introduced into the flow cytometer and the stained cells and beads fluoresce when excited by a laser beam.

The three-color reagent permits identification of lymphocyte subsets using fluorescence gating instead of forward scatter gating. This three-color reagent allows direct gating on the CD45positive population using a combination of fluorescence and side scatter parameters. By gating on the CD45-positive population, a maximum number of lymphocytes may be captured in the gate and non-lymphocyte contamination may be minimized.

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

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

Not Found

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

Performance of the product was established by testing at Cleveland Clinic, Johns Hopkins I enomance of the problem was events of North Carolina, and at Becton I rospital, Institute or ometry Systems laboratories in San Jose, California.

Several studies were performed:

Accuracy was determined by comparison to Simultest/ADCC. Accuracy data demonstrated the Tril EST / TRUCOUNT product's equivalence to Simultest/ADCC.
Use of an Isotype Control was studied. Data was analyzed for percent T lymphocytes and Ose of an 1990 per Souther, was control to set quadrant markers and then using only the stained sample to set quadrant markers. Data indicated that the reagent may be used with or without an isotype control.
Reference range studies were performed. Many variables such as age, sex and geographical location may influence the reference range. Each site must determine its own reference range.
A stability study was conducted to assess the time effect relating to age of blood (timefrom-draw) and the time effect relating to the age of the s-ain (time-from-sample preparation), as well as the combined effect of both. Stability was determined for both percentages and absolute counts. Results indicate that for absolute counts, samples should be stained and analyzed within 6 hours of draw.
Within-specimen reproducibility was performed at BDIS; 10 replicates from 1 high, 1 medium, and 1 low (with respect to T and B lymphocytes) were assessed. Within-specimen reproducibility was also performed at 3 clinical sites; 3 aliquots from each donor (n=92 for CD3+ and n=80 for CD19+) were assessed. Results demonstrated acceptable within-sample reproducibility.
Lincarity was determined using blood samples from 3 normal donors diluted to 5 concentrations, ranging from 16,700 to 200 lymphocytes/uL and from 31,000 to 2,500 WBC/uL. Results indicate a linear response over this range.
Cross reactivity of these clones is reported in the literature. Conjugation and product formulation have not changed their specificity.
Results from a goss platform reproducibility study indicated that TriTEST CD3 FITC/CD19 PE/CD45 PerCP with or without TRUCOUNT tubes may be used with flow cytometers not made by Becton Dickinson.

The results of the clinical studies demonstrate that the device is as safe and effective as the predicate device.

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.

K913192, K950342

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

Attachment F

K9 70742
Oct. 22, 1997

Summary of Safety and Effectiveness

Submitter Information (21 CFR 807,92(a)(1))

Submitter:	Becton Dickinson Immunocytometry Systems
2350 Qume Drive
San Jose, CA 95131-1807

Anna Longwell, Esq. Contact: Director, Regulatory Affairs - Corporate (408) 954-2254
Summary date: February 26, 1997

Name of Device and Classification (21 CFR 807.92(a)(2))

Becton Dickinson TriTEST™ reagent CD3 FITC/CD19 PE/CD45 Name: PerCP; TRUCOUNT TM Absolute Count Tubes
Class II Classification:

Predicate Device (21 CFR 807.92(a)(3))

The BDIS TriTEST™ CD3 FITC/CD19 PE/CD45 PerCP reagent with TRUCOUNT Absolute Count Tubes is substantially equivalent to Simultest™ IMK-Lymphocyte Tube C (cleved to market under 510(k) K913192) plus ADCC. TriTEST reagent CD3/CD19/CD45, when used to enumerate percentages of T and B lymphocytes was cleared to market under 510(k) K950342.

Description of the Device (21 CFR 807.92(2)(4))

306

Summary of Safety and Effectiveness

be identified and the events in this region counted. The proportion of reagent positive events to be identified and are trated. The absolute count is P x (beads/pellet)/(volume of blood sample).

Intended Use (21 CFR 807.92(a)(5))

For in vitro diagnostic use to identify and enumerate percentages and absolute counts of T and B lymphocytes in bload.

Indications for Use

For use with any flow cytometer with specified detection ranges .
For use with erythrocyte lysed whole blood .
For use with or without an isotype control .
For in vitro diagnostic use .
To identify and enumerate percentages and absolute counts of CD3+ and CD19+ ● lymphocytes
To characterize and monitor some forms of immunodeficiency .
To characterize and monitor some forms of autoimmune diseases .

Clinical Utility

The enumeration of T (CD3+) and B (CD19+) lymphocytes has been found useful in monitoring some forms of immunodeficiency and autoimmune disease.

Comparison to Predicate Device (21 CFR 807.92(a)(6))

The three-color reagent is substantially equivalent to the predicate: Simultest™ IMK-Lymphocyte Tube C plus hematology analyzer in that they share the same intended uses. The Simultest portion of the predicate and the TriTEST reagent use essentially the same monoclonal antibody/flow cytometric methodology. The TriTEST and Simultest products differ in the steps used to determine analysis gates to identify the lymphocyte population. The TriTEST /TRUCOUNT system compared to the predicate, requires less sample handling, a simpler sample preparation scheme and fewer sample tubes to achieve the same end result, Results demonstrate that the products yield essentially equivalent performance characteristics.

307

Summary of Safety and Effectiveness

Performance Data (21 CFR 807.92(b)(2))

Several studies were performed:

· Accuracy was determined by comparison to Simultest/ADCC. Accuracy data demonstrated the Tril EST / TRUCOUNT product's equivalence to Simultest/ADCC.
· Use of an Isotype Control was studied. Data was analyzed for percent T lymphocytes and Ose of an 1990 per Souther, was control to set quadrant markers and then using only the stained sample to set quadrant markers. Data indicated that the reagent may be used with or without an isotype control.
· Reference range studies were performed. Many variables such as age, sex and geographical location may influence the reference range. Each site must determine its own reference range.
· A stability study was conducted to assess the time effect relating to age of blood (timefrom-draw) and the time effect relating to the age of the s-ain (time-from-sample preparation), as well as the combined effect of both. Stability was determined for both percentages and absolute counts. Results indicate that for absolute counts, samples should be stained and analyzed within 6 hours of draw.
· Within-specimen reproducibility was performed at BDIS; 10 replicates from 1 high, 1 medium, and 1 low (with respect to T and B lymphocytes) were assessed. Within-specimen reproducibility was also performed at 3 clinical sites; 3 aliquots from each donor (n=92 for CD3+ and n=80 for CD19+) were assessed. Results demonstrated acceptable within-sample reproducibility.
Lincarity was determined using blood samples from 3 normal donors diluted to 5 concentrations, ranging from 16,700 to 200 lymphocytes/uL and from 31,000 to 2,500 WBC/uL. Results indicate a linear response over this range.
· Cross reactivity of these clones is reported in the literature. Conjugation and product formulation have not changed their specificity.
Results from a goss platform reproducibility study indicated that TriTEST CD3 FITC/CD19 PE/CD45 PerCP with or without TRUCOUNT tubes may be used with flow cytometers not made by Becton Dickinson.

Performance Data - Conclusions (21 CFR 807.92(b)(3))

The results of the clinical studies demonstrate that the device is as safe and effective as the predicate device.

CD3/CD19/CD45 TRUCOUNT 510(k) Notification Attachment F: Summary of Safety and Effectiveness

308
/

DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/3/Picture/2 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle with its wings spread, enclosed within a circle. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged around the upper portion of the circle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Anna Longwell, Esq. Director, Regulatory Affairs - Corporate Becton Dickinson Immunocytometry Systems 2350 Qume Drive San Jose, CA 95131-1807

OCT 2 2 1997

Re : K970742 Becton Dickinson TriTEST™ Reagent CD3 FITC/CD19 Trade Name: PE/CD45 PerCP; TruCOUNT™ Absolute Count Tubes Regulatory Class: II Product Code: GKZ Dated: August 29, 1997 Received: September 02, 1997

Dear Ms. Longwell:

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 Current Good Manufacturing Practice requirement, 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 for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. go 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) Additionally, for questions on the promotion and 594-4588. 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

| of_ Page_

K910742 (if known):-

dons For Use:

For use with any flow cytometer with specified detection ranges
t

For use with erythrocyte lysed whole blood

For use with or without an isotype control

For in vitro diagnostic use

For in viris cases of absolute counts of CD3+ and absolute counts of CD3+ and CD19+ lymphocytes

SE of the program of monitor some forms of immunodeficiency

To characterize and monitor some forms of autoimmune diseases .

IF YOU STILL NEED MORE SPACE ON THIS LINE, CONTINUE ON ANOTHER PAGE IF NEEDED).

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Peter E. Marin

(Division Sign-Of
Division of Clinical Lang
510(k) Number

Over-The-Counter Use_

Prescription Use Or 21 CFR 801.109)

(Optional Format 1-2-96).