For in vitro diagnostic use to identify and enumerate absolute counts of T lymphocyte (CD3+), helper/inducer T lymphocyte (CD3+CD4+) and suppressor/cytotoxic T lymphocyte (CD3+CD8+) subsets in blood.

For use with any flow cytometer equipped with a 488 nm last and capable of detection in . the ranges: 515-545 nm, 562-607 nm, and >650 nm
For use with erythrocyte lysed whole blood .
For in vitro diagnostic use .
To identify and enumerate absolute counts of CD3+ and CD3+CD4+ and CD3+CD8+ . lymphocytes
To characterize and monitor some forms of immunodeficiency, such as in HIV infected . individuals
To characterize and monitor some forms of autoimmune diseases .

Device Description

The BDIS TriTEST CD4 fluorescein isothiocyanate (FITC)/CD8 phycoerythrin (PE)/CD3 peridinin chlorophyll protein (PerCP) reagent is a three-color, direct immunofluorescence reagent. When used with TRUCOUNT Absolute Count Tubes it is used for identifying and enumerating absolute counts of I lymphocytes (CD3+), helper/inducer T lymphocytes (CD3+CD4+) and suppressor/cytotoxic I lymphocytes (CD3+CD8+) in erythrocyte-lysed whole blood (LWB). The Becton Dickinson TnTEST / TRUCOUNT system for immunophenotyping consists of a flow cytometer (either from BDIS or from another manufacturer), conjugated monoclonal reagent (Tril EST CD4 FITC/CD8 PE/CD3 PeCP) and TRUCOUNT Absolute Count Tubes.

The process to obtain T lymphocyte subset absolute counts includes: 1) obtaining a whole blood sample, 2) adding a precise volume of whole blood directly to the Absolute Count Tube, 3) cell-surface antigen staining with three-color monoclonal antibody reagents, 4) erythrocyte lysis, and 5) flow cytometric acquisition and analysis of list mode data. Analysis for absolute counts requires that the bead 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 I x (beads/pellet)/(volume of blood sample).

AI/ML Overview

Here's an analysis of the provided text, extracting the requested information about the acceptance criteria and the study that proves the device meets them:

Acceptance Criteria and Device Performance Study for BDIS TriTEST™ Reagent with TRUCOUNT Absolute Count Tubes

This submission does not explicitly state specific numerical acceptance criteria for performance metrics. Instead, it relies on demonstrating substantial equivalence to a predicate device (FACSCount cleared under K933486). The "acceptance criteria" are therefore inferred as achieving performance equivalent to or demonstrably safe and effective as the predicate device for the specified indications for use.

The study aims to establish this equivalence, particularly for identifying and enumerating absolute counts of T lymphocytes (CD3+), helper/inducer T lymphocytes (CD3+CD4+), and suppressor/cytotoxic T lymphocytes (CD3+CD8+).

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric	Acceptance Criteria (Inferred)	Reported Device Performance
Accuracy	Substantial equivalence to the predicate device (FACSCount) for CD3+, CD3+CD4+, and CD3+CD8+ absolute counts.	Accuracy data demonstrated the TriTEST/TRUCOUNT product's equivalence to FACSCount. "Results demonstrate that the products yield essentially equivalent performance characteristics." The TriTEST/TRUCOUNT system (for absolute counts) is substantially equivalent to the FACSCount system for CD3+, CD3+CD4+ and for CD3+CD8+.
Stability (Time-from-draw)	Blood specimens should be stained within a specified timeframe to maintain accurate absolute counts.	Blood specimens should be stained within 24 hours of draw. (Alternatively, stained within 48 hours of draw and analyzed within 6 hours).
Stability (Time-from-sample preparation)	Stained samples should be analyzed within a specified timeframe to maintain accurate absolute counts.	Analysis of stained samples should occur within 24 hours. (Alternatively, stained within 48 hours of draw and analyzed within 6 hours).
Within-specimen Reproducibility	Acceptable consistency in results when multiple aliquots from a single specimen are tested.	"Results demonstrated acceptable within-sample reproducibility." (Tested with 10 replicates from 1 high, 1 medium, and 1 low sample at BDIS, and 3 aliquots from each donor at 3 clinical sites).
Linearity	Linear response across a clinical range of lymphocyte and WBC concentrations.	"Results indicate a linear response over this range." (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).
Cross-reactivity	Monoclonal antibodies should maintain their specificity after conjugation and formulation.	"Conjugation and product formulation have not changed their specificity." (Cross-reactivity of clones is reported in literature).
Cross-platform Reproducibility	Acceptable comparability of results across different flow cytometer platforms.	Indicated a small (<20%) non-zero bias, but good correlation between results on a Becton Dickinson flow cytometer versus a Coulter. Users will be advised to validate performance characteristics as required under CLIA regulations. (Implies that while there's a bias, the correlation is good enough to allow for validation and use, with appropriate user validation).

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

Accuracy Study:
- Sample Size: Not explicitly stated, but the study was conducted at multiple sites: Cleveland Clinic, Johns Hopkins Hospital, Institute of Tropical Medicine, University of North Carolina Hospital, and Becton Dickinson Immunocytometry Systems laboratories. The number of patients/samples isn't given.
- Data Provenance: Retrospective and/or Prospective (not specified, but clinical studies are generally prospective or involve fresh samples). Origin is USA (Cleveland Clinic, Johns Hopkins, UNC Hospital, BDIS San Jose) and potentially other countries (Institute of Tropical Medicine - country not specified).
Stability Study (Time-from-draw/Stain): Not specified.
Within-specimen Reproducibility:
- BDIS site: 3 samples (1 high, 1 medium, 1 low) with 10 replicates each (total 30 replicates).
- 3 clinical sites: Number of donors/samples not specified, but 3 aliquots from each donor were assessed.
Linearity Study: Blood samples from 3 normal donors. Each donor's sample was diluted to 5 concentrations (15 total diluted samples tested).
Cross-platform Reproducibility: Not specified.

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

The document does not provide information about the number or qualifications of experts used to establish ground truth for the test set. Given the nature of a 510(k) for an in vitro diagnostic reagent, the "ground truth" would typically come from existing, FDA-cleared laboratory methods, often the predicate device itself, or established laboratory standards.

4. Adjudication Method for the Test Set

The document does not specify any adjudication method for the test set. For IVD reagents, the comparison is usually quantitative between the new device and the predicate device, or against a gold standard method, rather than involving expert consensus for individual case adjudication.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, and Effect Size

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This type of study is typically for image-based diagnostic systems where human readers interpret results, and the AI assists that interpretation. This device is a reagent/system for flow cytometry, which produces quantitative results, not images for human interpretation in the same way. Therefore, "human readers improve with AI vs. without AI assistance" is not applicable here.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done

This device is effectively a standalone algorithm/system in terms of its primary function. The flow cytometer with the reagent and TRUCOUNT tubes generates quantitative data (absolute counts) for cell populations. While a trained lab technician operates the flow cytometer and analyzes the data, the core "performance" of generating the counts is inherent to the reagent system itself. The studies described (accuracy, reproducibility, linearity) assess this "algorithm only" type of performance, comparing its output to that of the predicate device.

7. The Type of Ground Truth Used (Expert Consensus, Pathology, Outcomes Data, etc.)

The ground truth used for these studies is primarily:

Comparison to a Predicate Device: The main ground truth is the performance of the legally marketed FACSCount system (K933486). The studies aim to show the new device yields "essentially equivalent performance characteristics" and is "substantially equivalent" to this established device.
Internal Validation/Standard Methods: For reproducibility, linearity, and stability, the ground truth is established through controlled laboratory experiments using known samples or dilutions, comparing results against expected values or consistency metrics.
Literature: For cross-reactivity, it refers to existing literature on the specificity of the monoclonal antibody clones.

8. The Sample Size for the Training Set

The document does not mention a training set in the context of an algorithm or AI model development. This device is a reagent used in a flow cytometry system, not an AI model that requires a distinct training phase.

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

As there is no mention of a training set for an AI model, this question is not applicable. The device's "training" in a broad sense would be the extensive research and development (R&D) and validation of the reagent formulation and system, not an AI-specific training phase with labeled data.

Summary

{0}------------------------------------------------

K971205

Attachment F

Summary of Safety and Effectiveness

DEC - I

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

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

Anna Longwell, Esq. Contact: Director, Regulatory Affairs - Corporate (408) 954-2254

Summary date: March 31, 1997

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

Becton Dickinson Tril EST TM reagent CD4 FITC/CD8 PE/CD3 PecP; Name: TRUCOUNTIM Absolute Count Tubes
Classification: Class II

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

The BDIS TriTEST™ CD4 FITC/CD8 PE/CD3 PerCP reagent with TRUCOUNT Absolute Count Tubes is substantially equivalent to FACSCount (cleared to market under K933486).

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

{1}------------------------------------------------

Summary of Safety and Effectiveness

When monoclonal antibody reagents are added to human whole blood, the fluorochrome-In held 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 T lymphocyte subsets using fluorescence gating instead of forward scatter gating. This three-color reagent allows direct gating on the CD3+ population using a combination of fluorescence and side scatter parameters. By gating on the CD3+ population, a maximum number of T lymphocytes may be captured in the gate.

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

For in vitro diagnostic use to identify and enumerate absolute counts of T lymphocyte (CD3+), helper/inducer T lymphocyte (CD3+CD4+) and suppressor/cytotoxic T lymphocyte (CD3+CD8+) subsets in blood.

Indications for Use

For use with any flow cytometer equipped with a 488 nm last and capable of detection in . the ranges: 515-545 nm, 562-607 nm, and >650 nm
For use with erythrocyte lysed whole blood .
For in vitro diagnostic use .
To identify and enumerate absolute counts of CD3+ and CD3+CD4+ and CD3+CD8+ . lymphocytes
To characterize and monitor some forms of immunodeficiency, such as in HIV infected . individuals
To characterize and monitor some forms of autoimmune diseases .

Clinical Utility

The determination of T lymphocyte (CD3+), helper/inducer T lymphocyte (CD3+CD4+) and suppressor/cytotoxic T lymphocyte (CD3+CD8+) has been found useful in monitoring some forms of immunodeficiency and autoimmune disease.

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

The TriTEST /TRUCOUNT system (for absolute counts) is substantially equivalent to the FACSCount system for CD3+, CD3+CD4+ and for CD3+CD8+, (cleared to market under K933486). Both the TriTEST reagent with TRUCOUNT tubes and the predicate device yield equivalent results for the same analytes, and both are intended for use as an in vitro diagnostic test using a flow cytometer-based instrument and recommended computer hardware and software. Results demonstrate that the products yield essentially equivalent performance characteristics.

CD4/CD8/CD3/T RUCKINT 510(k) Notification Attachment F: Summary of Safety and Effectiveness

{2}------------------------------------------------

ﺔ ﺍﻟﻤ

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

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

Several studies were performed:

Accuracy data demonstrated the TriTEST/TRUCOUNT product's equivalence to ● FACSCount.
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. Results indicate that for the calculation of absolute counts, blood specimen should be stained within 24 hours of draw, and analysis of the stained samples should occur within 24 hours or alternatively, the blood specimen can be stained within 48 hours of draw and analyzed within 6 hours.
Within-specimen reproducibility was performed at BDIS; 10 replicates from 1 high, 1 . medium, and 1 low were assessed. Within-specimen reproducibility was also performed at 3 clinical sites; 3 aliquots from each donor were assessed. Results demonstrated acceptable within-sample reproducibility.
. Linearity 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 cross platform reproducibility study for determining absolute counts using . TriTEST CD4 FITC/CD8 PE/CD3 PerCP reagent with TRUCOUNT Absolute Count Tubes indicated a small (<20%) non-zero bias, but good correlation between results on a Becton Dickinson flow cytometer versus a Coulter. Therefore, users will be advised that they must validate performance characteristics for absolute counts, as required under CLIA regulations (42 CFR 493.1202 and 493.1213).

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.

CD4/CD8/CD3/TRUCOUNT 510(k) Notification Attachment F: Summary of Safety and Effectiveness

{3}------------------------------------------------

Image /page/3/Picture/2 description: The image is a black and white seal for the Department of Health & Human Services - USA. The seal is circular with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract image of an eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Anna Longwell Director, Regulatory Affairs BECTON DICKINSON IMMUNOCYTOMETRY SYSTEMS 2350 Qume Drive San Jose, CA 95131-1807

DEC - 1 1997

Re: K971205 Trade Name: Becton Dickinson TriTEST Reagent CD4 FITC/CD8 PE/CD 3 PerCP; TruCOUNT Absolute Count Tubes Regulatory Class: II Product Code: GKZ, 81 Dated: March 31, 1997 April 1, 1997 Received:

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 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. this response to your premarket notification Please note: 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.

{4}------------------------------------------------

Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To 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) 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 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

{5}------------------------------------------------

Page 1 of /

510(k) Number (if known): K 971205

Device Name:

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 ervthrocyte lysed whole blood
� For in vitro diagnostic use
To identify and enumerate absolute counts of CD3+ and CD3+CD4 and � CD3+CD8+ lymphocytes
� To characterize and monitor some forms of immunodeficiency, such as in HIV infected individuals
◆ To characterize and monitor some forms of autoimmune diseases

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Peter E. Maher

Oivision Sign-Off) Division of Clinical Laboratory Devices Laboratory Devices Laboratury Devices Laboratory Devices Laboratory Devices Laboratory Devices Laboratory December 510(k) Number

Prescription Use
(Per 21 CFR 801.109) √

Over-The-Counter Use__________________________________________________________________________________________________________________________________________________________

(Optional Format 1-2-96)

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.”