For use with Becton Dickinson FAC® flow cytometers equipped with a blue (488-nm) and red diode (635-nm) laser.
Monoclonal antibody reagents for identification and enumeration of mature human lymphocyte subsets in peripheral blood of normal individuals and patients with certain immune dysfunction.
MultiTEST CD3/CD16+56/CD45/CD19 provides percentages and absolute counts of T (CD3'), NK(CD3 CD16'CD56'), and B (CD3'CD19') lymphocytes.
MultiTEST CD3/CD8/CD45/CD4 provides percentages and absolute counts of T (CD3*), helper/inducer T (CD3CD4) and suppressor/cytotoxic T (CD3CD8) lymphocytes.
For use with erythrocyte lysed whole blood without an isotype control.
To characterize and monitor forms of auto immune diseases, such as lupus.
To characterize and monitor congenital or acquired immunodeficiences, such as SCID or AIDS.
For in vitro diagnostic use

Device Description

The Becton Dickinson CD3 fluorescein isothiocyanate (FITC)/CD16+56 phycoerythrin (PE)/CD45 peridinin chlorophyll protein (PerCP)/CD19 allophycocyanin (APC) reagent is a four-color, direct immunofluorescence reagent for identifying and enumerating percentages and absolute counts of CD3+ T lymphocytes, CD3-CD16+56+ natural killer (NK), and CD3-CD19+ B lymphocyte subsets in erythrocyte-lysed whole blood. Each vial of reagent yields 50 tests. The reagent is intended for use on the Becton Dickinson FACSort™ or FACSCalibur™ flow cytometers equipped with the FL4 Option, the Apple® Macintosh® Quadra or PowerPC computer, and CELLQuest™ or MultiSET™ software. Daily instrument set-up requires CaliBRITE™ beads (unlabeled, FITC, PE, PerCP, and APC) and FACSComp™ software. The four-color method permits the identification and enumeration of T, NK, and B lymphocyte subsets using MultiSET software's expert lymphocyte gate and automated single-tube quality control algorithms. A lymphocyte gate is drawn for the CD45 * leucocytes with low side scatter and the lymphocyte subsets are provided as an absolute count or percentage of total lymphocytes.

AI/ML Overview

This document describes the performance data for the Becton Dickinson MultiTEST CD3/CD16+56/CD45/CD19 and MultiTEST IMK Kit, which are flow cytometry reagents used for identifying and enumerating lymphocyte subsets. The goal of the studies was to demonstrate substantial equivalence to predicate devices (TriTEST CD3/CD16+56/CD45 and TriTEST CD3/CD19/CD45).

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are not explicitly stated with specific numerical thresholds (e.g., a certain percentage agreement or coefficient of variation). Instead, the studies conclude that the performance is "acceptable" or "equivalent" to the predicate devices. The reported device performance is based on these qualitative assessments.

Performance Metric	Acceptance Criteria (Implicit)	Reported Device Performance
Accuracy	Equivalent to predicate devices	MultiTEST CD3/CD16+56/CD45/CD19 is equivalent to TriTEST CD3/CD16+56/CD45 and TriTEST CD3/CD19/CD45.
Within-site Reproducibility	Acceptable	Demonstrated acceptable within-site reproducibility.
Across-site Reproducibility	Acceptable	Demonstrated acceptable across-site reproducibility.
Stability	Acceptable	Demonstrated acceptable stability of samples prepared up to 48 hours after blood draw and up to 24 hours after preparation.
Linearity and Recovery	Linear response and acceptable recovery	Results indicate linear response over the tested range and acceptable recovery.

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

Accuracy Study:
- Sample Size: 129 lysed whole blood (LWB) specimens.
  - 70 normal donors
  - 59 abnormal donors
- Data Provenance: The study was conducted at two clinical sites, indicating prospective collection from human subjects. The country of origin is not specified but implied to be the US given the FDA submission.
Within-site Reproducibility:
- Sample Size: LWB specimens from 3 normal and 6 abnormal donors (total 9 donors). Each specimen was divided into 10 aliquots.
- Data Provenance: Not explicitly stated but likely prospective data from human subjects.
Across-site Reproducibility:
- Sample Size: LWB specimens from a total of 16 normal and 30 abnormal donors (total 46 donors). Each specimen was divided into 5 aliquots.
- Data Provenance: Conducted at three clinical sites, indicating prospective collection from human subjects.
Stability Studies:
- Sample Size: Samples from 10 normal and 20 abnormal donors (total 30 donors).
- Data Provenance: Not explicitly stated but likely prospective data from human subjects.
Linearity and Recovery:
- Sample Size: Blood specimens from 3 normal donors.
- Data Provenance: Not explicitly stated but likely prospective data from human subjects.

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

The document does not describe the use of human experts to establish "ground truth" in the traditional sense (e.g., radiologists interpreting images). For flow cytometry reagents, the "ground truth" and comparison are typically established by comparing the results of the new device to a legally marketed predicate device (which itself would have established its performance against other methods or clinical outcomes), or by established laboratory reference methods. In this case, the test set's ground truth for accuracy was based on the performance of the predicate devices. The "normal" and "abnormal" classifications of the donor samples would likely be based on clinical diagnosis or established laboratory ranges, but no explicit expert involvement in defining the ground truth for each individual sample's classification is mentioned beyond the comparison to the predicate.

4. Adjudication Method for the Test Set

Not applicable. The "adjudication" in this context refers to the comparison of the new device's results with those of the predicate device, or established reference methods in a laboratory setting. There is no mention of a human adjudication process by multiple experts for discrepancies in interpretations.

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 not a study involving human readers interpreting medical images or data with or without AI assistance. It's a performance study of diagnostic reagents used in flow cytometry.

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

This study evaluates the performance of a diagnostic reagent kit in conjunction with specific flow cytometers and software (Becton Dickinson FACSort or FACSCalibur flow cytometers, Apple Macintosh computers, CELLQuest™ or MultiSET™ software). While the software includes "expert lymphocyte gate and automated single-tube quality control algorithms," the overall system involves human operation and interpretation of the flow cytometer output. Therefore, it is not a "standalone algorithm only" study in the sense of a fully automated AI system without human interaction. The performance is of the device (reagent kit + specific instrumentation/software) as an integrated system in a laboratory setting.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)

The primary "ground truth" for the accuracy study was the results obtained from the predicate devices (TriTEST CD3/CD16+56/CD45 and TriTEST CD3/CD19/CD45). The study aimed to demonstrate "equivalence" to these already-marketed and accepted devices. For the other studies (reproducibility, stability, linearity), the "ground truth" would be inherent in the expected biological or technical performance for those assays, often against established laboratory controls or expected ranges. The classification of samples as "normal" or "abnormal" would be based on clinical criteria and laboratory values but not necessarily a separate "expert consensus" on the specific flow cytometry results themselves for the purpose of validating the device.

8. The Sample Size for the Training Set

The document does not provide information on a "training set" as it would apply to a machine learning algorithm. This submission describes the validation of a laboratory reagent and associated instrumentation/software, where the "algorithms" (e.g., for gating) are likely rule-based or pre-programmed, rather than learned from a large, labeled training dataset.

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

Not applicable, as a distinct "training set" and associated ground truth establishment process, typical for machine learning models, is not described for this device. The software's "expert lymphocyte gate and automated single-tube quality control algorithms" would have been developed based on scientific understanding of cell populations and validated through internal R&D processes, rather than a single, distinct "training set" in the AI sense.

Summary

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MAY 22 1998

510(k) Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21CFR807.92.

The assigned 510(k) number is_

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

Submitter:	Becton Dickinson Immunocytometry Systems2350 Qume DriveSan Jose, CA 95131-1807
Contact:	Cindy Morrow

Sr. Regulatory Specialist (408) 954-2694 (408) 954-2495 - Fax Cindy Morrow@bdis.com

Summary date: March 4, 1998

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

Name:	MultiTEST CD3/CD16+56/CD45/CD19 and MultiTEST IMK Kit
Classification:	Class II Device

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

MultiTEST CD3/CD16+56/CD19, TRUCOUNT absolute count tubes, and MultiTEST IMK Kit are substantially equivalent to TriTEST CD3/CD16+56/CD45 (K971110), TriTEST CD3/CD19/CD45 (K970742), TRUCOUNT tubes (K970836), and MultiTEST CD3/CD8/CD45/CD4 reagent (974360).

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

The Becton Dickinson CD3 fluorescein isothiocyanate (FITC)/CD16+56 phycoerythrin (PE)/CD45 peridinin chlorophyll protein (PerCP)/CD19 allophycocyanin (APC) reagent is a four-color, direct immunofluorescence reagent for identifying and enumerating percentages and absolute counts of CD3+ T lymphocytes, CD3-CD16+56+ natural killer (NK), and CD3-CD19+ B lymphocyte subsets in erythrocytelysed whole blood. Each vial of reagent yields 50 tests. The reagent is intended for use on the Becton Dickinson FACSort™ or FACSCalibur™ flow cytometers equipped with the FL4 Option, the Apple® Macintosh® Quadra or PowerPC computer, and CELLQuest™ or MultiSET™ software. Daily instrument set-up requires CaliBRITE™ beads (unlabeled, FITC, PE, PerCP, and APC) and FACSComp™ software. The four-color method permits the identification and enumeration of T, NK, and B lymphocyte subsets using MultiSET software's expert lymphocyte gate and automated single-tube quality control algorithms. A lymphocyte gate is drawn for the CD45 * leucocytes with low side scatter and the lymphocyte subsets are provided as an absolute count or percentage of total lymphocytes.

* The term "substancial equivalence" as used in this 510(k) notification of substantial equivalence as found in the Federal Food, Drug, and Cosmeric Act, as amended and as applied under 21 CFR 807, Subpart E under which a device can be marketed without pre-marker approval or reclassification of substantial equivalency under this notification is not intended to have any bearing whatsoever on the resolution of patent infingement suits or any other patent naters. No statements related to, or in support of substance herein shall be construed as an admission against interest under the US Patent Laws or their application by the courts.

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

MultiTEST CD3/CD16+56/CD45/CD19 reagent is a four-color reagent for identifying and enumerating percentages and absolute counts of T (CD3+), NK (natural killer) (CD3+CD16+56+), and B (CD3+CD19+) lymphocyte subsets by direct immunofluorescence. The MultiTEST IMK contains the same MultiTEST CD3/CD16+56/CD45/CD19 reagent, MultiTEST CD3/CD8/CD45/CD4 reagent, and MultiTEST IMK Kit Lysing Solution. Helper/inducer T (CD3+CD4+) and suppressor/cytoroxic T (CD3+CD8+) lymphocyte subser percentages and absolute counts can be obtained from the MultiTEST CD3/CD8/CD45/CD4 reagent included in the kit. Subsets of lymphocytes are useful in managing some forms of immunodeficiency diseases.

Technological Characteristics (21 CFR 807.92(a)(6))

MultiTEST CD3/CD16+56/CD45/CD19 is a four-color reagent and TriTEST CD3/CD16+56/CD45 and TriTEST CD3/CD19/CD45 are three-color reagents. The three and four-color reagents all employ the same monoclonal antibody clones, however the CD19 antibody is conjugated to PE in the three-color reagent and to APC in the four-color reagent. The three-color reagent includes FITC, PE, and PerCP fluorescent dyes that are excited by the flow cytometer's blue (488-nm) argon-ion laser. The four-color reagent contains an additional APC fluorescent dye. The APC is excited by a red diode (635-nm) laser provided in Becton Dickinson's FL4 Option for the FACSort and FACSCalibur flow cytometers.

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

Performance of the product was established by testing at four centers.

Several studies were performed:

· Accuracy was determined by comparing results from 129 lysed whole blood (LWB) specimens, including 70 normals and 59 abnormals at two clinical sites. Accuracy data demonstrated that MultiTEST CD3/CD16+56/CD45/CD19 is equivalent to TriTEST CD3/CD16+56/CD45 and TriTEST CD3/CD19/CD45.
Within-site reproducibility was performed on LWB specimens from 3 normal and 6 abnormal donors. Each specimen was divided into 10 aliquots and then stained and analyzed using a FACSCalibur system. Results demonstrated acceptable within-site reproducibility.
· Across-site reproducibility was performed at three clinical sites on LWB specimens from a total of 16 normal and 30 abnormal donors. Each specimen was divided into 5 aliquots and then stained and analyzed using a FACSCalibur system. Results demonstrated acceptable across-site reproducibility.
Stability studies were conducted at one site using samples from 10 normal and 20 abnormal donors. Results demonstrate acceptable stability of samples prepared up to 48 hours after blood draw and stability of prepared samples up to 24 hours after preparation.

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Linearity and recovery were determined using blood specimens from 3 normal donors diluted to 5 different concentrations, ranging from 200 to 29,700 WBC/uL and from 100 to 9000 lymphocytes/pL. Results indicate linear response over this range and acceptable recovery.
Comparison to Predicate Device (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. The four-color MultiTEST CD3/CD16+56/CD45/CD19 reagent is substantially equivalent to the predicate three-color TriTEST CD3/CD16+56/CD45 and TriTEST CD3/CD19/CD45 reagents, in that they share the same intended uses and methodologies. Results demonstrate that the products yield essentially equivalent performance characteristics.

Cindy Morrow

March 4, 1998

Date

Sr Regulatory Specialist

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MAY 222 1998

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Cindy Morrow Sr. Regulatory Specialist BECTON DICKINSON IMMUNOCYTOMETRY SYSTEMS 2350 Oume Drive San Jose, CA 95131-1807

Re: K980858

Trade Name: MultiTEST CD3/CD16+56/CD45/CD19 and MultiTEST IMK Kit Regulatory Class: II Product Code: GKZ Dated: March 04, 1998 Received: March 05, 1998

Dear Ms. Morrow:

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 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 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. 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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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 Sutman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

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Indications for Use Statement

510(k) Number:

5980858

Device Name:

MultiTEST CD3/CD16+56/CD45/CD19 and MultiTEST IMK kit

For use with Becton Dickinson FAC® flow cytometers equipped with a blue (488-nm) . and red diode (635-nm) laser.
Monoclonal antibody reagents for identification and enumeration of mature human . lymphocyte subsets in peripheral blood of normal individuals and patients with certain immune dysfunction.
MultiTEST CD3/CD16+56/CD45/CD19 provides percentages and absolute counts of T . (CD3'), NK(CD3 CD16'CD56'), and B (CD3'CD19') lymphocytes.
MultiTEST CD3/CD8/CD45/CD4 provides percentages and absolute counts of T . (CD3*), helper/inducer T (CD3CD4) and suppressor/cytotoxic T (CD3CD8) lymphocytes.
For use with erythrocyte lysed whole blood without an isotype control. .
To characterize and monitor forms of auto immune diseases, such as lupus. .
To characterize and monitor congenital or acquired immunodeficiences, such as SCID or � AIDS.
. For in vitro diagnostic use

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Peter E. Makim

Perscription Use
(Per 21 CFR 801.109)

Over-The-Counter Use _

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.”