K Number

Device Name

TRITEST REAGENT CD3 FITC/CD4 PE/CD45 PERCP; TRUCOUNT ABSOLUTE COUNT TUBES

Manufacturer

BECTON DICKINSON IMMUNOCYTOMETRY SYSTEMS

Date Cleared

1997-11-21

(338 days)

Product Code

GKZ

Regulation Number

864.5220

Intended Use

For in vitro diagnostic use to identify and enumerate percentages and absolute counts of CD3+ and CD3+CD4+ lymphocytes in blood. For use with any flow cytometer equipped with a 488 nm laser and capable of detection in the ranges: 515-545 nm, 562-607 nm, and > 650 nm. For use with erythrocyte-lysed peripheral whole blood. For use with or without an isotype control. To characterize and monitor some forms of autoimmune disease. To characterize and monitor some forms of immunodeficiency disease, such as in HIV-infected individuals.

Device Description

The BDIS TriTEST CD3 fluorescein isothiocyanate (FITC)/CD4 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 T-helper (CD3+CD4+) cells in erythrocyte-lysed whole blood (LWB). When used with TRUCOUNT Absolute Count Tubes, the product will yield absolute counts in cells/UL. The Becton Dickinson TriTEST/TRUCOUNT system for immunophenotyping consists of a flow cytometer (either from BDIS or from another manufacture), conjugated monoclonal reagent (TriTEST CD3 FITC/CD4 PE/CD45 PerCP) 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+CD4+) 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, be identified and the events in this region counted. The proportion of reagent positive events to bead events (P) is computed. The absolute count is P x (beads/pellet)/(volume of blood sample). When monoclonal antibody reagents are added to human whole blood, the fluorochrome-labeled antibodies bind specifically to antigens on the surface of leukocytes, thus identifying lymphocyte populations. The patient blood sample is added to the counting bead pellet 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

K933486

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on traditional flow cytometry methods, antibody binding, and manual gating/analysis based on fluorescence and scatter parameters. There is no mention of automated feature extraction, pattern recognition, or learning algorithms typically associated with AI/ML in this context.

Therapeutic

No
The device is an in vitro diagnostic (IVD) tool designed to identify and quantify specific lymphocyte populations (CD3+ and CD3+CD4+) in blood for diagnostic and monitoring purposes, not for direct treatment or therapy.

Diagnostic

Yes

The device is explicitly stated to be "For in vitro diagnostic use" and its intended uses include "characterize and monitor some forms of autoimmune disease" and "characterize and monitor some forms of immunodeficiency disease, such as in HIV-infected individuals," which are diagnostic purposes.

SaMD (Software as a Medical Device)

The device description clearly outlines hardware components (reagent, tubes, flow cytometer) and a physical process involving blood samples, staining, lysis, and flow cytometry. It is not solely software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states "For in vitro diagnostic use".
Device Description: The device is a reagent used to identify and enumerate cell populations in blood samples, which is a common application for in vitro diagnostics.
Clinical Context: The intended use includes characterizing and monitoring autoimmune and immunodeficiency diseases, which are clinical applications.
Performance Studies: The performance studies described (accuracy, reference range, stability, reproducibility, linearity) are typical for validating an IVD device.
Predicate Device: The mention of a predicate device (BDIS FACSCount system) further indicates that this device is intended for a similar diagnostic purpose and regulatory pathway.

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 CD3+ and CD3+CD4+ lymphocytes in blood.

For use with any flow cytometer equipped with a 488 nm laser and capable of detection ♥ in the ranges: 515-545 nm, 562-607 nm, and > 650 nm
For use with erythrocyte-lysed peripheral whole blood .
For use with or without an isotype control .
To characterize and monitor some forms of autoimmune disease .
To characterize and monitor some forms of immunodeficiency disease, such as in HIV-infected . individuals

Product codes

GKZ

Device Description

The BDIS TrilleST CD3 flucescein isothiogranate (FITC)/CD4 phycoerythrin (PE)/CD45 peridinin chlorophyll protein (PeCP) reagent is a three-color, direct immunofluqescence reagent for identifying and enumerating percentages of T lymphocytes (CD3+) and T-helper (CD3+CD4+) cells in aythrocyte-lysed whole blood (LWB). When used with TRUCOUNT Absolute Count Tubes, the product will yield absolute counts in cells/UL. The Becton Dickinson TrillEST/TRUCOUNT system for immunophenotyping consists of a flow cytomere (cither from BDIS or from another manufacture), conjugated monoclonal reagent (TriTEST CD3 FITC/CD4 PE/CD45 PerCP) 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) cythrocyte lysis, and 4) flow cytometric acquisition and analysis of list mode data. Analysis involves computing the ratio of reagent-positive events (CD3+CD4+) 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, be identified and the events in this region counted. The proportion of reagent positive wents to bead events (P) is computed. The absolute count is I' x (beads/peller)/(volume of blood sample).

When monoclonal antibody reagents are added to human whole blood, the flucrochrome-labeled antibodies bind specifically to antigens on the surface of leucocytes, thus identifying lymphogye populations. The patient blood sample is added to the counting bead pellet and is treated with fluctochrome-labeled antibodies and the cythrocytes are lysed with FACS® Lysing Solution. The flow cytomerer 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 cyrometer 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 flucescence and side scatter parameters. By gating on the CD45positive population, a maximum number of lymphocytes may be captured in the gate and nonlymphocyte 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

Accuracy: determined by comparison to the FACSCount system. Absolute counts for CD3+ were obtained from 197 (152 abnormal and 45 normal) dones. Absolute counts for CD3+CD4+ were obtained from dones; data for CD3+CD4+ was grouped according to range of number of cells. Accuray data demonstrated the TriTEST/TRUCOUNT product's equivalence to FACSCount. A subset of the accuracy study data was analyzed for percent CD3+ and percent CD4+ first using a control to set matkers and then using only the stained sample to set quadrant markers. Data indicated that the reagent may be used with or without an isorype control.
Reference range studies: performed. The reference range for CD3+ and CD3+CD4+ lymphocytes was a function of sex and site. For CD3+ lymphocytes, within the male population, it was also a function of age. Each site must determine its own reference range.
Stability study: conducted to assess the time effect relating to age of blood (time-from-draw) and the time effect relating to the age of the stain (time-from-sample preparation), as well as the combined effect of both. Stability was decermined for both percentages and absolute counts. Stability was decemined for whole blood samples at 0, 24, 48 and 72 hours post draw. Additionally, stability was measured for stained samples at 0 and 24 hours from time of staining. A combination of the two, time from draw plus time from sample preparation was srudied. Results indicated that either 1) staining the samples within 24 hours of daw and analyzing samples within 24 hours of staining or alternatively, 2) staining the samples with 48 hours of draw and analyzing them within 6 hours is recommended.
Within-specimen reproducibility: performed at BDIS; 10 replicates from 1 high, 1 medium, and 1 low (with respect to CD3+ and CD3+CD4+ lymphocytes) were assessed. Performed at 3 clinical sites; 3 aliquots from each donor (n-94 for CD3+ and n=80 for CD3+CD4+) were assessed. Results demonstrated acceptable within-sample reproducibility.
Linearity: determined using blood samples from 3 normal dones diluted to 5 concentrations, ranging from 16,700 to 200 lymphoorcs/JLL 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.
Cross platform reproducibility study: indicated 1) for percentage results the reagent may be used with flow cytometers not made by Becton Dickinson 2) for determining absolute counts usingTriTEST reagent with TRUCOUNT Absolute Count Tubes, there was a small (

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

K9605053

Attachment C

Summary of Safety and Effectiveness

NOV 21 1997

Submitter Information (21 CFR 807.92(2)(1))

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

Anna Longwell, Esq. Contact: Director, Regulatory Affairs - Corporate (408) 954-2254
Summary date: November 18, 1997

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

Becton Dickinson TriTEST24 reagent CD3 FITC/CD4 PE/CD45 PerCP; Name: TRUCOUNTTM Absolute Count Tubes

Classificacion: Class II

Predicate Device (21 CFR 807.92(=1(3))

The BDIS TrillEST™ CD3/CD4/CD45 reagent with TRUCOUNT Absolute Count Tubes is substantially equivalent to the BDIS FACSCount system cleared in K933486.

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

Summary of Safety and Effectiveness

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

For in vitro diagnostic use to identify and enumerate percentages and absolute counts of CD3+ and CD3+CD4+ lymphocytes in blood.

Indications for Use

For use with any flow cytometer equipped with a 488 nm laser and capable of detection ♥ in the ranges: 515-545 nm, 562-607 nm, and > 650 nm
For use with erythrocyte-lysed peripheral whole blood .
For use with or without an isotype control .
To characterize and monitor some forms of autoimmune disease .
To characterize and monitor some forms of immunodeficiency disease, such as in HIV-infected . individuals

Clinical Urility

The determination of CD3+CD4+ lymphocytes has been found useful in monitoring some forms of immunodeficiency and autoimmune disease.

Summary of Safety and Effectiveness

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

The three-color reagent is substantially equivalent to the predicate BDIS FACSCount System in that they share the same intended uses and similar methodologies. Results demonstrate that the products yield essentially equivalent performance characteristics. Both tests are antibody-dependent, in vitro diagnostic assays to identify T lymphocytes (CD3+) and helper/induce (CD3+CD4+) Tlymphocyte subsets. The products differ in the steps used to determine analysis gates to identify the lymphocyte population.

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

Performance of the product was established by testing at Cleveland Clinic, Johns Hopkins Hospitul, Institute of Tropical Medicine, University of North Carolina, and at Becton Dickinson Immunocytometry Systems laboratories in San Jose, California.

Several studies were performed:

Accuracy was determined by comparison to the FACSCount system. Absolute counts for CD3+ t were obtained from 197 (152 abnormal and 45 normal) dones. Absolute counts for CD3+CD4+ were obtained from dones; data for CD3+CD4+ was grouped according to range of number of cells. Accuray data demonstrated the TriTEST/TRUCOUNT product's equivalence to FACSCount. Additionally, a subser of the accuracy study data was analyzed for percent CD3+ and percent CD4+ first using a control to set matkers and then using only the stained sample to set quadrant markers. Data indicated that the reagent may be used with or without an isorype control.
Reference range studies were performed. The reference range for CD3+ and CD3+CD4+ . lymphocytes was a function of sex and site. For CD3+ lymphocytes, within the male population, it was also a function of age. Each site must determine its own reference range.
A stability study was conducted to assess the time effect relating to age of blood (time-from-. draw) and the time effect relating to the age of the stain (time-from-sample preparation), as well as the combined effect of both. Stability was decermined for both percentages and absolute counts. Stability was decemined for whole blood samples at 0, 24, 48 and 72 hours post draw. Additionally, stability was measured for stained samples at 0 and 24 hours from time of staining. A combination of the two, time from draw plus time from sample preparation was srudied. Results indicated that either 1) staining the samples within 24 hours of daw and analyzing samples within 24 hours of staining or alternatively, 2) staining the samples with 48 hours of draw and analyzing them within 6 hours is recommended.

Summary of Safety and Effectiveness

Within-soccimen reproducibility was performed at BDIS; 10 replicates from 1 high, 1 medium, and 1 low (with respect to CD3+ and CD3+CD4+ lymphocytes) were assessed. Withinspecimen reproducibility was also performed at 3 clincial sites; 3 aliquots from each donor (n-94 for CD3+ and n=80 for CD3+CD4+) were assessed. Results demonstrated acceptable withinsample reproducibility.
Linearity was determined using blood samples from 3 normal dones diluted to 5 . concentrations, ranging from 16,700 to 200 lymphoorcs/JLL 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 cross platform reproducibility study indicated 1) for percentage results the reagent . may be used with flow cytometers not made by Becton Dickinson 2) for determining absolute counts usingTriTEST reagent with TRUCOUNT Absolute Count Tubes, there was a small ( 650 nm
For use with erythrocyte-lysed peripheral whole blood .
For use with or without an isotype control .
To characterize and monitor some forms of autoimmune diseasc .
To characterize and monitor some forms of immunodeficiency disease, such as in . HIV-infected individuals

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Artu E. Makeri

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

Prescription Use (Per 21-CFR 801.109) OR

Over-The-Counter Use

(Optional Format 1-2-96)