The BD FACSLyric™ flow cytometer is intended for use as an in vitro diagnostic device for immunophenotyping using up to six fluorescence detection channels and two light scatter channels using a blue (488-nm) and a red (640-nm) laser. It is intended for use with in vitro diagnostic (IVD) assays and software that are indicated for use with the instrument.

BD Multitest™ 6-color TBNK, BD Multitest™ IMK kit, BD Multitest™ CD3/CD8/CD45/CD4, and BD Multitest™ CD3/CD16+CD56/CD45/CD19, all with optional BD Trucount™ tubes, are intended for use on the BD FACSLyric flow cytometer with peripheral whole blood for immunophenotyping. These reagents are indicated for use in the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency. These reagents determine the percentages and absolute counts of the following mature human lymphocyte subsets:

BD Multitest 6-color TBNK with optional BD Trucount tubes

· T lymphocytes (CD3+)
· B lymphocytes (CD19+)
· Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
· Helper/inducer T lymphocytes (CD3+CD4+)
· Suppressor/cytotoxic T lymphocytes (CD3+CD8+)

BD Multitest IMK kit with optional BD Trucount tubes

· T lymphocytes (CD3+)
· B lymphocytes (CD19+)
· Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
· Helper/inducer T lymphocytes (CD3+CD4+)
· Suppressor/cytotoxic T lymphocytes (CD3+CD8+)

BD Multitest CD3/CD8/CD45/CD4 with optional BD Trucount tubes

· T lymphocytes (CD3+)
· Suppressor/cytotoxic T lymphocytes (CD3+CD8+)
· Helper/inducer T lymphocytes (CD3+CD4+)

BD Multitest CD3/CD16+CD56/CD45/CD19 with optional BD Trucount tubes

· T lymphocytes (CD3+)
· Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
· B lymphocytes (CD3-CD19+)

Device Description

The BD FACSLyric Flow Cytometer consists of the following components.

FACSLyric Flow Cytometer (3-1, 4-2, 4-2-2 and 4-3-3 optical configurations)
FACSuite Clinical Software
Multitest 6-Color Assay Modules
Multitest 4-Color Assay Modules
FC Beads 7-Color Kit
CS&T Beads
Multitest 6-Color TBNK
Multitest IMK Kit
. Multitest CD3/CD8/CD45/CD4
Multitest CD3/CD16+CD56/CD45/CD19
. Multi-Check Control
. Multi-Check CD4 Low Control
. Trucount Tubes
Optional FACS Universal Loader

The system is an immunofluorescence assay system for identification and enumeration of lymphocyte subsets in peripheral whole blood. When blood is added to the monoclonal antibody reagent, the fluorochrome-labeled antibodies in the reagent bind specifically to lymphocvte surface antigens. During acquisition, the cells travel past one or more laser beams in a single file and scatter the laser light. The stained cells fluoresce. The scatter and fluorescence light is detected by the flow cytometer, separated by dichroic mirrors and optical filters, and then quantified with photomultiplier tubes (PMTs) to determine the percent lymphocyte of particular cell populations.

Multiple monoclonal antibodies each labeled with a different fluorochrome are contained within one reagent tube to simultaneously identify multiple lymphocyte subset populations. The use of Trucount tubes provides the absolute number of the fluorochrome-labeled antibody bound cells.

AI/ML Overview

The provided text describes the BD FACSLyric™ Flow Cytometer, its indications for use, comparison to a predicate device (BD FACSCanto™ II system), and performance data supporting its substantial equivalency. Based on the provided document, here's an analysis of the acceptance criteria and study proving device performance:

Important Note: This document is an FDA 510(k) summary, which focuses on demonstrating substantial equivalence to a legally marketed predicate device rather than strictly presenting a clinical study design with explicit acceptance criteria for a novel AI/ML device. Therefore, some information (e.g., number of experts, adjudication methods, MRMC study effect size, sample size for training set, ground truth for training set) typically available for AI/ML device studies may not be directly available or applicable in this context. The study described focuses on demonstrating the performance of a new flow cytometer instrument rather than an AI/ML algorithm.

1. Table of Acceptance Criteria and Reported Device Performance

The document defines acceptance criteria primarily through "Objectives" for each study and "Results" indicating whether these objectives were met. The acceptance criteria are largely based on meeting performance requirements that demonstrate equivalency to the predicate device or established clinical laboratory guidelines (CLSI standards).

Study Name	Objective (Acceptance Criteria)	Reported Device Performance (Results)
Bench Performance:
Intra-instrument Optical Configuration Equivalency	To demonstrate system level performance equivalency across the 3-1, 4-2, 4-2-2, and 4-3-3 optical configurations as measured by QC and setup performance parameters generated utilizing CS&T beads and FC beads.	System level optical equivalency was demonstrated between the four configurations.
Inter-instrument Optical Configuration Equivalency	To demonstrate performance equivalency between the 3-1, 4-2, 4-2-2, and 4-3-3 optical configurations of the FACSLyric through testing multiple instruments of different configurations (inter-instrument) using patient and normal donor samples stained with Multitest IMK kit and Multitest 6-Color TBNK in Trucount tubes.	The performance equivalency of the 6 IVD channels between the 3-1, 4-2, 4-2-2 and 4-3-3 optical configurations of the FACSLyric flow cytometer have been demonstrated.
Method Comparison	- To determine the method bias between the FACSLyric system and the FACSCanto II system on the determination of CD3+, etc., percentage and absolute counts (meeting acceptance criteria for bias). - To demonstrate equivalency between FACSLyric Universal Loader (UL) acquisition and FACSLyric manual acquisition.	The method bias between the FACSLyric and the FACSCanto II met the acceptance criteria. Equivalency between FACSLyric UL acquisition and Manual acquisition was demonstrated.
Within-site Precision	To evaluate the within-site precision performance of FACSLyric system using Multitest IMK kit and Multitest 6-Color TBNK with Trucount tubes (meeting within-run and total precision requirements).	All acceptance criteria were met. The within-site precision performance satisfied the within-run and total precision performance requirements.
Whole Blood Repeatability	To verify repeatability performance across all FACSLyric instruments of different configurations, using whole blood from patient and normal donor samples, stained with reagents (all lymphocyte subsets meeting %CV and SD acceptance criteria).	Repeatability performance was demonstrated across all instruments with different configurations. All lymphocyte subsets met %CV and SD acceptance criteria.
Linearity	To evaluate the linear range of the FACSLyric system using reagents (acceptable linear ranges established for each lymphocyte subset).	Acceptable linear ranges have been established for each lymphocyte subset for each of the Multitest reagents on FACSLyric system.
Sample Carryover	To determine percent sample carryover (less than or equal to 0.1% for low carryover and less than or equal to 0.5% for standard carryover).	All results met the acceptance criteria (less than or equal to 0.1% for low carryover and less than or equal to 0.5% for standard carryover) on all three FACSLyric instruments. Single SIT flush sufficient for low carryover.
Reagent Carryover	To verify minimal reagent carryover.	Results met the acceptance criteria. Amount of reagent carried over is well below the amount required for effective staining.
Detection Capability of LoB and LoD	To evaluate the limit of blank (LoB) and limit of detection (LoD) (LoB and LoD for CD4 lymphocyte subset absolute counts less than 50 cells/µL).	LoB and LoD for CD4 lymphocyte subset absolute counts were established and were less than 50 cells/µL.
Detection Capability of LoQ	To evaluate the limit of quantitation (LoQ) (LoQ for CD4 lymphocyte subset absolute counts less than 50 cells/µL).	The LoQ for the CD3+, CD3+CD4+, CD3+CD8+, CD19+, CD16+CD56+ and CD45+ lymphocyte subsets were established. Acceptance criteria were met and the LOQ is less than 50 cells/µL for CD4 lymphocyte subset absolute counts.
Setup and QC Performance	To verify that Setup & QC Performance meets pre-defined specifications.	All Setup & QC performance results met the acceptance criteria.
Setup and QC Stability	To verify the stability of system level Setup & QC.	All system level Setup & QC stability performance results met the pre-defined specifications.
Clinical Performance:
Method Comparison	To evaluate performance equivalence of the FACSLyric system and the FACSCanto II system on the determination of lymphocyte subsets.	The acceptance criteria for all lymphocyte subsets were met for the FACSLyric system. Differences were minimal and no bias trend observed when comparing manipulated vs non-manipulated specimens or different EDTA formulations.
Inter-laboratory Reproducibility	To evaluate inter-laboratory reproducibility for the FACSLyric system (variability for within run, between run, between day, between site, and total precision meeting 97.5% one-sided confidence interval compared to requirements).	Inter-laboratory reproducibility was carried out at four clinical sites. Variability met requirements and 97.5% one-sided confidence interval on precision was estimated and compared.
Reference Intervals	To re-establish the reference intervals of a normal male and female adult cohort (reference intervals established for all lymphocyte subsets).	Reference Intervals for all lymphocyte subsets were established. Differences observed between genders were not clinically significant.
Specimen Stability	To generate data to support stability of venous whole blood with EDTA anticoagulant when testing reagents on the FACSLyric system (supporting stability claims for Age of Blood and Age of Stain).	The results support the claim of up to 24 hours Age of Blood and 6 hours Age of Stain for Multitest 6-Color TBNK, and up to 48 hours Age of Blood and 24 hours Age of Stain for Multitest IMK kit (including other Multitest assays) on FACSLyric system.

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

Inter-instrument Optical Configuration Equivalency & Whole Blood Repeatability: Used patient and normal donor samples from unspecified locations.
Method Comparison (Bench Study): Samples sourced from unspecified locations.
Method Comparison (Clinical Study): A total of 332 specimens were enrolled for Multitest 6-Color TBNK (297 evaluable), and 368 specimens were enrolled for Multitest IMK kit (336 evaluable) at 5 clinical sites. These were remnant, de-linked patient specimens. The provenance is not explicitly stated as retrospective or prospective, but "remnant, de-linked patient specimens" typically implies a retrospective use of existing samples.
Inter-laboratory Reproducibility: Used a single lot of Streck CD-Chex Plus process control material at four clinical sites. (Not a human patient sample size).
Reference Intervals: A total of 136 subjects were enrolled (134 evaluable for Multitest 6-Color TBNK and 130 evaluable for Multitest IMK kit) from one clinical site. These were prospectively procured and de-linked donor specimens.
Specimen Stability: A total of 227 specimens were enrolled for each of the reagents from two clinical sites. These were prospectively procured and de-linked patient specimens.

Data Provenance (General): While specific countries are not mentioned, the document is an FDA submission for the US market, implying that the studies were conducted in accordance with US regulatory requirements, likely involving US-based clinical sites. The data included both normal donor and patient samples, and were both prospectively and retrospectively collected (e.g., remnant, de-linked specimens).

3. Number of Experts Used to Establish Ground Truth and Qualifications:

This document describes a flow cytometer and associated reagents, not an AI/ML algorithm that requires expert labeling of images or complex data for ground truth. The "ground truth" for this type of device is the accurate measurement and enumeration of cell subsets, which is compared against results from a predicate device or validated laboratory methods. Therefore, the concept of "experts establishing ground truth for the test set" as typically understood in AI/ML validation studies (e.g., radiologists annotating images) is not directly applicable here. The accuracy of the cell counts would be intrinsically tied to the performance of the predicate device (BD FACSCanto II system) and the established clinical laboratory guidelines (CLSI standards) used for comparison.

4. Adjudication Method for the Test Set:

Not applicable in the context of this device and study type. The studies involve quantitative measurements and comparisons against established parameters or a predicate device, rather than subjective interpretations requiring adjudication among multiple readers.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:

No, an MRMC comparative effectiveness study, as typically done for AI/ML devices assisting human readers in diagnostic tasks, was not conducted or described. This submission relates to a flow cytometer instrument, where performance is measured by its ability to accurately count and phenotype cells, rather than its effect on human interpretations of complex data like medical images.

6. Standalone (Algorithm Only) Performance:

Not applicable. The device is a "Flow Cytometer," an automated instrument for in vitro diagnostic use. Its performance is inherently "standalone" in executing its function of cell counting and phenotyping. There is no separate "algorithm-only" component as might be found in an AI/ML software. The software (FACSuite Clinical Software) manages the instrument and data analysis for the user, and its performance is evaluated as part of the overall system.

7. Type of Ground Truth Used:

The ground truth is established by:

Predicate Device Comparison: The performance of the BD FACSLyric™ system is compared to a legally marketed predicate device, the BD FACSCanto™ II system. This is a primary form of "ground truth" for demonstrating substantial equivalence.
Established Clinical Laboratory Standards: Adherence to various Clinical and Laboratory Standards Institute (CLSI) guidelines (e.g., CLSI EP09-A3 for Method Comparison and Bias Estimation, CLSI EP05-A3 for Precision, CLSI EP6-A for Linearity, CLSI H26-A2 for Carryover, CLSI EP17-A2 for Detection Capability, CLSI EP28-A3c for Reference Intervals) serves as the "ground truth" for acceptable analytical performance. These standards provide accepted methodologies and performance metrics for laboratory assays.
Quantitative Measurement: The device measures percentages and absolute counts of lymphocyte subsets. The "ground truth" for these measurements is the true biological count in the sample, which is assessed through rigorous validation encompassing precision, linearity, limits of detection, and comparison to a well-characterized predicate.

8. Sample Size for the Training Set:

Not applicable. This is not an AI/ML device, and there is no mention of a "training set" for an algorithm. The development and validation of flow cytometers rely on engineering principles, analytical studies, and clinical performance studies, not on training data for a machine learning model.

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

Not applicable, as there is no training set for an AI/ML algorithm.

Summary

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Image /page/0/Picture/1 description: The image shows the seal of the Department of Health & Human Services - USA. The seal is circular and contains the department's name around the perimeter. In the center of the seal is an abstract symbol that resembles a stylized caduceus or a series of human profiles.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

August 28, 2017

Becton, Dickinson and Company Li Zhou Manager, Regulatory Affairs, BD Biosciences 2350 Qume Drive San Jose. CA 95131

Re: K170974

Trade/Device Name: BD FACSLyric™ Flow Cytometer Regulation Number: 21 CFR 864.5220 Regulation Name: Automated differential cell counter Regulatory Class: II Product Code: OYE Dated: March 31, 2017 Received: April 03, 2017

Dear Ms. Zhou:

This letter corrects our substantially equivalent letter of July 3, 2017.

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and 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) that do not require approval of a premarket approval application (PMA). 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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements

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as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If vou desire specific advice for your device on our labeling regulation (21 CFR Part 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely,

Leonthena R. Carrington -S

Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K170974

Device Name

BD FACSLyric™ Flow Cytometer

Indications for Use (Describe)

BD Multitest 6-color TBNK with optional BD Trucount tubes

· T lymphocytes (CD3+)
· B lymphocytes (CD19+)
· Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
· Helper/inducer T lymphocytes (CD3+CD4+)
· Suppressor/cytotoxic T lymphocytes (CD3+CD8+)

BD Multitest IMK kit with optional BD Trucount tubes

· T lymphocytes (CD3+)
· B lymphocytes (CD19+)
· Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
· Helper/inducer T lymphocytes (CD3+CD4+)
· Suppressor/cytotoxic T lymphocytes (CD3+CD8+)

BD Multitest CD3/CD8/CD45/CD4 with optional BD Trucount tubes

· T lymphocytes (CD3+)
· Suppressor/cytotoxic T lymphocytes (CD3+CD8+)
· Helper/inducer T lymphocytes (CD3+CD4+)

BD Multitest CD3/CD16+CD56/CD45/CD19 with optional BD Trucount tubes

· T lymphocytes (CD3+)
· Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
· B lymphocytes (CD3-CD19+)

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Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

| Over-The-Counter Use (21 CFR 801 Subpart C)

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Section 5 510(k) Summary

This bundled 510(k) summary is being provided in accordance with the requirements of the Safe Medical Devices Act of 1990 and 21 CFR 807.92.

Date of Summary: March 31, 2017

5.1 Submitted By

BD Biosciences 2350 Oume Drive San Jose, CA 95131-1807 USA

Contact:	Li ZhouManager, Regulatory Affairs
Telephone:	(408) 954-2099
Fax:	(408) 954-2347
Email:	li.zhou2@BD.com

5.2 Device

Trade Name/Device Name: BD FACSLyric™ Flow Cytometer

Classification: Class II Device Classification: Flow Cytometric Reagents and Accessories Regulation Description: Automated Differential Cell Counter Regulation Medical Specialty: Hematology Product Code: OYE Regulation Number: 21 CFR 864.5220

5.3 Predicate Device

The predicate device is BD FACSCanto™ II system with BD Multitest™ reagents that consists of the following:

FACSCanto II flow cytometer (4-2, 4-2-2 and 5-3 optical configurations) (K141468 ● and K062087)
. FACSCanto Clinical Software (Multitest 6-Color and 4-Color assays are included as panels within FACSCanto Clinical software) (K141468 and K062087)
FACS 7-Color Setup Beads (K040026) ●
Multitest 6-Color TBNK (K090967)
Multitest IMK Kit (K980858)
Multitest CD3/CD8/CD45/CD4 (K974360)
Multitest CD3/C16+CD56/CD45/CD19 (K980858)
Multi-Check Control (K961610) ●
Multi-Check CD4 Low Control (K982231) ●
Trucount Tubes (K970836)
Optional FACS Loader (K141468 and K062087)

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5.4 Device Description

The BD FACSLyric Flow Cytometer consists of the following components.

FACSLyric Flow Cytometer (3-1, 4-2, 4-2-2 and 4-3-3 optical configurations) ●
FACSuite Clinical Software ●
Multitest 6-Color Assay Modules ●
Multitest 4-Color Assay Modules ●
FC Beads 7-Color Kit
CS&T Beads
Multitest 6-Color TBNK
Multitest IMK Kit
. Multitest CD3/CD8/CD45/CD4
Multitest CD3/CD16+CD56/CD45/CD19 ●
. Multi-Check Control
. Multi-Check CD4 Low Control
. Trucount Tubes
Optional FACS Universal Loader ●

In accordance with FDA Guidance for Industry and FDA Staff, Bundling Multiple Devices or Multiple Indications in a Single Submission, dated June 22, 2007, four Multitest assays with the instrument, beads and process controls are bundled within this submission.

The FACSLyric System with Multitest Reagents is intended for the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency using flow cytometry applications.

The FACSLyric System with Multitest Reagents is intended for Prescription Use Only and will be labeled "Rx Only".

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5.5 Indications for Use

5.6.1 BD FACSLvric Flow Cytometer

BD Multitest™ 6-color TBNK, BD Multitest™ IMK kit, BD Multitest™ CD3/CD8/CD45/CD4. and BD Multitest™ CD3/CD16+CD56/CD45/CD19. all with optional BD Trucount™ tubes, are intended for use on the BD FACSLyric flow cytometer with peripheral whole blood for immunophenotyping. These reagents are indicated for use in the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency. These reagents determine the percentages and absolute counts of the following mature human lymphocyte subsets:

BD Multitest 6-color TBNK with optional BD Trucount tubes

T lymphocytes (CD3+) ●
B lymphocytes (CD19*) .
. Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56*)
. Helper/inducer T lymphocytes (CD3CD4)
Suppressor/cytotoxic T lymphocytes (CD3CD8)

BD Multitest IMK kit with optional BD Trucount tubes

T lymphocytes (CD3+) .
. B lymphocytes (CD19*)
Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56*)
Helper/inducer T lymphocytes (CD3+CD4*) .
Suppressor/cytotoxic T lymphocytes (CD3CD8) .

BD Multitest CD3/CD8/CD45/CD4 with optional BD Trucount tubes

T lymphocytes (CD3+)
Suppressor/cytotoxic T lymphocytes (CD3+CD8+) .
. Helper/inducer T lymphocytes (CD3+CD4+)

BD Multitest CD3/CD16+CD56/CD45/CD19 with optional BD Trucount tubes

T lymphocytes (CD3+) ●
Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
B lymphocytes (CD3-CD19+) .

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5.6.2 BD FC beads 7-color kit

The BD™ FC beads 7-color kit (BD FC beads), in conjunction with BD FACSuite™ Clinical software and BD™ CS&T beads, are used to establish fluorescence compensation for the BD FACSLyric™ flow cytometer.

5.6.3 BD CS&T beads

BD™ CS&T beads, in conjunction with BD FACSuite™ Clinical software, provide a standardized method for the quality control of optics, electronics, and for adjusting detector voltages and fluorescence compensation on the BD FACSLyric™ flow cytometer.

BD Multitest 6-color TBNK 5.6.4

BD Multitest™ 6-color TBNK with optional BD Trucount™ tubes is intended for use with BD FACSLyric™, BD FACSCanto™ II, and BD FACSCanto™ flow cytometers to determine the percentages and absolute counts of the following mature human lymphocyte subsets in peripheral whole blood for immunophenotyping:

T lymphocytes (CD3+) ●
. B lymphocytes (CD19*)
Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56*)
Helper/inducer T lymphocytes (CD3+CD4+)
Suppressor/cytotoxic T lymphocytes (CD3CD8) ●

This reagent is indicated for use in the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency.

BD Multitest IMK Kit 5.6.5

BD Multitest™ IMK kit with optional BD Trucount™ tubes is intended for use with BD FACSLyric™, BD FACSCanto™ II, BD FACSCanto™ , and BD FACSCalibur™ flow cytometers to determine the percentages and absolute counts of the following mature human lymphocyte subsets in peripheral whole blood for immunophenotyping:

T lymphocytes (CD3+) .
B lymphocytes (CD19+) ●
Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
. Helper/inducer T lymphocytes (CD3+CD4+)
Suppressor/cytotoxic T lymphocytes (CD3+CD8+) ●

This reagent is indicated for use in the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency.

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5.6.6 BD Multitest CD3/CD8/CD45/CD4

BD Multitest™ CD3/CD8/CD45/CD4 with optional BD Trucount™ tubes is intended for use with BD FACSLyric™, BD FACSCanto™ II, BD FACSCanto™, and BD FACSCalibur™ flow cytometers to determine the percentages and absolute counts of the following mature human lymphocyte subsets in peripheral whole blood for immunophenotyping:

T lymphocytes (CD3+) ●
. Helper/inducer T lymphocytes (CD3+CD4+)
Suppressor/cvtotoxic T 1vmphocytes (CD3+CD8+) .

This reagent is indicated for use in the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency.

BD Multitest CD3/CD16+CD56/CD45/CD19 5.6.7

BD Multitest™ CD3/CD16+CD56/CD45/CD19 with optional BD Trucount™ tubes is intended for use with BD FACSLyric™, BD FACSCanto™ II, BD FACSCanto™, and BD FACSCalibur™ flow cytometers to determine the percentages and absolute counts of the following mature human lymphocyte subsets in peripheral whole blood for immunophenotyping:

· T lymphocytes (CD3+)
Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+) .
B lymphocytes (CD3-CD19+) .

This reagent is indicated for use in the immunological assessment of normal individuals, and patients having, or suspected of having, immune deficiency.

5.6.8 BD Multi-Check Control

BD™ Multi-Check Control is intended as a complete process control for immunophenotyping by flow cytometry. It is a control for antibody staining, red blood cell (RBC) lysis, instrument setup and performance, and data analysis on BD FACSLyric™, BD FACSCanto™ II, BD FACSCanto™, and BD FACSCalibur™ flow cvtometers.

5.6.9 BD Multi-Check CD4 Low Control

BD™ Multi-Check CD4 Low Control is intended as a complete process control for immunophenotyping by flow cytometry. It is a control for antibody staining, red blood cell (RBC) lysis, instrument setup and performance, and data analysis on BD FACSLyric™, BD FACSCanto™ II, BD FACSCanto™, and BD FACSCalibur™ flow cytometers.

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5.6.10 BD Trucount Tubes

BD Trucount™ tubes are intended for use with appropriated BD IVD reagent products on BD FACSLyric™, BD FACSVia™, BD FACSCanto™, BD FACSCanto™ II and BD FACSCalibur™ flow cytometers to determine absolute counts of leucocytes in erythrocyte-lysed whole blood.

5.6 Comparison of Technological Characteristics with the Predicate Device

A comparison of the similarities and differences between the subject device and the predicate device is presented in Table 5-1 to Table 5-4

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	Table 5-1 Classification and Methodology
--	------------------------------------------	--	--

Feature/Attribute	Subject Device	Predicate Device
DeviceClassification andProduct Code	Automated Differential Cell Counter Regulatory Class: II Regulation Number: 21 CFR 864.5220 Product Code: OYE	Same
AssayMethodology	Flow Cytometry	Same
Detection/AssayPrinciple	Immunofluorescence	Same
MonoclonalAntibody Reagents	Multitest 6-color TBNK Multitest IMK kit Multitest CD3/CD8/CD45/CD4 Multitest CD3/CD16 +CD56/CD45/CD19 All with optional Trucount tubes to provide identification andenumeration of Total CD3+, CD3+CD4+, CD3+CD8+,CD16+CD56+ and CD19+ percentages and absolute countresults	Same
Specimen Type	Peripheral whole blood	Same
Sample Volume	50 µL	Same
SamplePreparation	Manual	Manual Automated (when used with optional accessory FACSSample Prep Assistant III )
Feature/Attribute	Subject Device	Predicate Device
Intended Use/	Flow Cytometer
Indications forUse - FlowCytometer	The BD FACSLyric flow cytometer is intended for use as an invitro diagnostic device for immunophenotyping using up to sixfluorescence detection channels and two light scatter channelsusing a blue (488-nm) and a red (640-nm) laser. It is intended foruse with in vitro diagnostic (IVD) assays and software that areindicated for use with the instrument.BD Multitest 6-color TBNK, BD Multitest IMK kit, BD MultitestCD3/CD8/CD45/CD4, and BD MultitestCD3/CD16+CD56/CD45/CD19, all with optional BD Trucounttubes, are intended for use on the BD FACSLyric flow cytometerwith peripheral whole blood for immunophenotyping. Thesereagents are indicated for use in the immunological assessment ofnormal individuals, and patients having, or suspected of having,immune deficiency. These reagents determine the percentagesand absolute counts of the following mature human lymphocytesubsets:BD Multitest 6-color TBNK with optional BD Trucount tubesT lymphocytes (CD3+)●B lymphocytes (CD19+)●Natural killer (NK) lymphocytes (CD3-CD16+ and/or●CD56+)Helper/inducer T lymphocytes (CD3+CD4+)●Suppressor/cytotoxic T lymphocytes (CD3+CD8+)●BD Multitest IMK kit with optional BD Trucount tubesT lymphocytes (CD3+)●	The BD FACSCanto II flow cytometers (4-2-2 and 5-3configurations) function as part of a system with dedicated clinicalsoftware intended for use with cleared or approved in vitrodiagnostic (IVD) assays that are indicated for use with theinstrument for the identification and enumeration of human cellsubsets. Only six detection channels using a blue (488 nm) and ared (633 nm) laser have been cleared for in vitro diagnostic use. Foruse with or without the BD FACS Sample Prep Assistant III.The BD FACSCanto II flow cytometer (4-2 configuration) isintended for use as an In Vitro Diagnostic device for identificationand enumeration of lymphocyte subsets in human cells insuspension.· Immunophenotyping in clinical laboratories, using previouslycleared in vitro diagnostic assays for flow cytometry.· Identification and enumeration of lymphocyte subsets in humancells in suspension.· For in vitro diagnostic use.· For use with or without the BD FACS Sample Prep Assistant IIISee intended use for BD Multitest reagents in Table 5-4.
	B lymphocytes (CD19+)●Natural killer (NK) lymphocytes (CD3-CD16+ and/or●CD56+)Helper/inducer T lymphocytes (CD3+CD4+)●Suppressor/cytotoxic T lymphocytes (CD3+CD8+)●
Feature/Attribute	Subject Device	Predicate Device
	BD Multitest CD3/CD8/CD45/CD4 with optional BD Trucounttubes
	• T lymphocytes (CD3+)
	• Suppressor/cytotoxic T lymphocytes (CD3+CD8+)
	• Helper/inducer T lymphocytes (CD3+CD4+)
	BD Multitest CD3/CD16+CD56/CD45/CD19 with optional BDTrucount tubes
	• T lymphocytes (CD3+)
	• Natural killer (NK) lymphocytes (CD3-CD16+ and/or CD56+)
	• B lymphocytes (CD3-CD19+)
MaximumParameterDetectors	IVD detection channels – Six Fluorescence channels plus forward scatter and side scatter	Same
Forward ScatterDetection	Photodiode with built-in 488/10 bandpass filter	Same
Fluorescence andSide ScatterDetection	Side scatter and fluorescence• Reflective optics with single transmission bandpass filter in front of each PMT• High performance PMT modules for all fluorescence and side scatter channels• Light collected by objective lens is delivered by fiber optics to specially designed detector arrays• The cuvette flow cell is gel-coupled by refractive index-matching optical gel to the fluorescence objective lens (1.2 NA) for optimal collection efficiency.	Same
Forward and SideScatter Sensitivity	Enables separation of fixed platelets from noise.	Same
SampleIdentification	Sample identification can be done through manual entry of sample information into the software or using an optional barcode reader.	Same
OpticalConfigurations	• 2-laser (blue, red), 4-color (3-1)• 2-laser (blue, red), 6-color (4-2)• 3-laser (blue, red, violet), 8-color (4-2-2)	• 2-laser (blue, red), 6-color (4-2)• 2 laser (blue, red), 8-color (5-3)• 3-laser (blue, red, violet), 8-color (4-2-2)
Feature/Attribute	Subject Device	Predicate Device
	3-laser (blue, red, violet), 10-color (4-3-3)
	Only up to six detection channels using red and blue lasers arefor IVD use.	Only up to six detection channels using red and blue
Lasers Used forIVD DetectionChannels	Blue Wavelength: 488 nm Optical power: 20 mW Red Wavelength: 640 nm Optical power: 40 mW	Blue Wavelength: 488 nm Optical power: 20 mW Red Wavelength: 633 nm Optical power: 17 mW
Electronics	Up to 35000 events/sec	Up to 10000 events/sec
SampleIntroduction	Manual loading onto the tube port of the flow cytometer Automated loading through a multi-tube FACS UniversalLoader (hold one 30 or 40-tube rack)	Manual loading onto the tube port of the flow cytometer Automated loading through a multi-tube FACS Loader (carousel tohold up to 40 tubes)
Software	FACSuite Clinical software with the following Multitest assaymodules: Multitest 6-Color Assays Multitest 4-Color Assays	FACSCanto Clinical software with Multitest 6-Color and Multitest4-Color assays included as panels within the software
Sample Analysis	Automated gating of cellular populations by the software andmanual adjustment by the user	Same
Results Reporting	Software- assisted report generation	Same
Fluidics	Uses FACSFlow as the sheath fluid Uses 10% bleach solution for system cleaning System waste products are collected in the waste container(5L standard size) Uses FACSFlow as the sheath fluid	Uses FACSFlow as the sheath fluid, together with FACSshutdown solution and FACS cleaning solution System waste products are collected in the waste container (10L standard size)

Table 5-2 Instrument and Software

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Feature/ Attribute	Subject Device	Predicate Device
	Quality Control and Setup Beads
Intended Use/Indications for Use	BD FC beads 7-color kitThe BD FC beads 7-color kit (BD FC beads), in conjunctionwith BD FACSuite Clinical software and BD CS&T beads, areused to establish fluorescence compensation for the BDFACSLyric flow cytometer.BD CS&T beadsBD CS&T beads, in conjunction with BD FACSuite Clinicalsoftware, provide a standardized method for the quality controlof optics, electronics, and fluidics, and for adjusting detectorvoltages and fluorescence compensation on the BD FACSLyricflow cytometer.	BD FACS 7-Color Setup BeadsFor in vitro diagnostic use on a BD FACSCanto flow cytometerwith BD FACSCanto software. The beads are used to adjustfluorescent detector voltages, to set fluorescence compensation, andto monitor daily instrument performance.
Quality Control &Instrument Setup	Daily QC performed using CS&T beads QC is also preformed every 6 months using CS&T beads.It includes all of the measurements performed in daily QCalong with additional more detailed measurements and anautomatic laser alignment. Daily instrument setup using CS&T beads FC Beads are run every 2 months to measure and resetinstrument spectral overlap values as part of the instrumentsetup	Daily QC performed using FACS 7-color setup Daily instrument setup using FACS 7-color setup beads

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Table 5-4 Monoclonal Antibody Reagents and Process Controls

Feature/Attribute	Subject Device	Predicate Device
	Multitest 6-Color TBNK
IntendedUse/Indicationsfor Use	BD Multitest 6-color TBNK with optional BD Trucount tubes isintended for use with BD FACSLyric, BD FACSCanto II, and BDFACSCanto flow cytometers to determine the percentages andabsolute counts of the following mature human lymphocyte subsetsin peripheral whole blood for immunophenotyping:T lymphocytes (CD3+) B lymphocytes (CD19+) Natural killer (NK) lymphocytes (CD3-CD16+ and/orCD56+) Helper/inducer T lymphocytes (CD3+CD4+) Suppressor/cytotoxic T lymphocytes (CD3+CD8+) This reagent is indicated for use in the immunological assessmentof normal individuals, and patients having, or suspected of having,immune deficiency.	BD Multitest 6-color TBNK reagent with optional BD Trucounttubes is a six-color direct immunofluorescence reagent for use withBD FACSCanto and BD FACSCanto II flow cytometers to identifyand determine the percentages and absolute counts of T, B, andnatural killer (NK) cells as well as the CD4 and CD8 subpopulationsof T cells in peripheral blood.BD Multitest 6-color TBNK reagent and BD Trucount tubes can beused with the BD FACS Loader.
Feature/Attribute	Subject Device	Predicate Device
	Multitest IMK Kit
IntendedUse/Indicationsfor Use	BD Multitest IMK kit with optional BD Trucount tubes is intendedfor use with BD FACSLyric, BD FACSCanto II, BD FACSCanto ,and BD FACSCalibur flow cytometers to determine thepercentages and absolute counts of the following mature humanlymphocyte subsets in peripheral whole blood forimmunophenotyping:• T lymphocytes (CD3+)• B lymphocytes (CD19+)• Natural killer (NK) lymphocytes (CD3-CD16+ and/orCD56+)• Helper/inducer T lymphocytes (CD3+CD4+)• Suppressor/cytotoxic T lymphocytes (CD3+CD8+)This reagent is indicated for use in the immunological assessmentof normal individuals, and patients having, or suspected of having,immune deficiency.	Intended UseMultiTESTCD3/CD16+56/CD45/CD19 reagent is a four-colorreagent for identifying and enumerating percentages and absolutecounts of T (CD3+), NK (natural killer) (CD3- CD16+CD56+), andB (CD3-CD19 +) lymphocyte subsets by directimmunofluorescence. The MultiTEST IMK contains the sameMultiTEST CD3/CD16+CD56/CD45/CD19) reagent, MultiTESTCD3/CD8/CD45/CD4 reagent, and MultiTEST IMK Kit LysingSolution. Helper/inducer T (CD3+CD4+) and suppressor/cytotoxic T(CD3+CD8+) lymphocyte subset percentages and absolute countscan be obtained from the MultiTEST CD3/CD8/CD45/CD4 reagentincluded in the kit. Subsets of lymphocytes are useful in managingsome forms of immunodeficiency diseases.Indications for Use• For use with Becton Dickinson FACS flow cytometers equippedwith a blue (488-nm) and red diode (635-nm) laser.• Monoclonal antibody reagents for identification andenumeration of mature human lymphocyte subsets in peripheralblood of normal individuals and patients with certain immunedysfunction.• MultiTEST CD3/CD16+56/CD45/CD19 provides percentagesand absolute counts of T (CD3+), NK (CD3-CD16+CD56+) andB (CD3-CD19+) lymphocytes. MultiTESTCD3/CD8/CD45/CD4 provides percentages and absolute countsof T (CD3+), helper/inducer T (CD3+CD4+) andsuppressor/cytotoxic T (CD3+CD8+) lymphocytes.• For use with erythrocyte lysed whole blood without an isotypecontrol.• To characterize and monitor forms of auto immune diseases,such as lupus.• To characterize and monitor congenital or acquiredimmunodeficiences, such as SCID or AIDS.For in vitro diagnostic use.
Feature/Attribute	Subject Device	Predicate Device
	Multitest CD3/CD8/CD45/CD4
IntendedUse/Indicationsfor Use	BD Multitest CD3/CD8/CD45/CD4 with optional BD Trucounttubes is intended for use with BD FACSLyric, BD FACSCanto II,BD FACSCanto, and BD FACSCalibur flow cytometers todetermine the percentages and absolute counts of the followingmature human lymphocyte subsets in peripheral whole blood forimmunophenotyping:T lymphocytes (CD3+)●Helper/inducer T lymphocytes (CD3+CD4+)●Suppressor/cytotoxic T lymphocytes (CD3CD8)●This reagent is indicated for use in the immunological assessmentof normal individuals, and patients having, or suspected of having,immune deficiency.	Intended UseMultiTEST CD3/CD8/CD45/CD4 reagent is a four-color reagent foridentifying and enumerating CD3+ T Lymphocytes, CD3+CD4+helper/inducer, and CD3+CD8+ suppresser/cytotoxic T Lymphocytesubsets by direct immunofluorescence. Subsets of T Lymphocytesare useful in managing immunodeficiency diseases. To characterizeand monitor congenital or acquired immunodeficiences, such asSCID or AIDS.Indications for UseFor the FACS family of flow cytometers equipped with a blue●(488-nm) and a red diode (635-nm) laser.A monoclonal antibody reagent for identification and●enumeration of mature human T lymphocyte subsets in humanperipheral blood.For use with erythrocyte lysed whole blood.●To characterize and monitor forms of autoimmune diseases,●such as lupus.To characterize and monitor congenital or acquired●immunodeficiences, such as SCID or AIDS.
	For in vitro diagnostic use.Multitest CD3/CD16+CD56/CD45/CD19
IntendedUse/Indicationsfor Use -MultitestReagents	BD Multitest CD3/CD16+CD56/CD45/CD19 with optional BDTrucount tubes is intended for use with BD FACSLyric, BDFACSCanto II, BD FACSCanto, and BD FACSCalibur flowcytometers to determine the percentages and absolute counts of thefollowing mature human lymphocyte subsets in peripheral wholeblood for immunophenotyping:T lymphocytes (CD3+)●Natural killer (NK) lymphocytes (CD3-CD16+ and/or●CD56+)B lymphocytes (CD3-CD19+)●	Intended UseMultiTESTCD3/CD16+56/CD45/CD19 reagent is a four-colorreagent for identifying and enumerating percentages and absolutecounts of T (CD3+), NK (natural killer) (CD3- CD16+CD56+), andB (CD3-CD19 +) lymphocyte subsets by directimmunofluorescence. The MultiTEST IMK contains the sameMultiTEST CD3/CD16+CD56/CD45/CD19) reagent, MultiTESTCD3/CD8/CD45/CD4 reagent, and MultiTEST IMK Kit LysingSolution. Helper/inducer T (CD3+CD4+) and suppresser/cytotoxic T(CD3+CD8+) lymphocyte subset percentages and absolute countscan be obtained from the MultiTEST CD3/CD8/CD45/CD4 reagent

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Feature/Attribute	Subject Device	Predicate Device
	This reagent is indicated for use in the immunological assessmentof normal individuals, and patients having, or suspected of having,immune deficiency.	included in the kit. Subsets of lymphocytes are useful in managingsome forms of immunodeficiency diseases.Indications for UseFor use with Becton Dickinson FAC flow cytometers equippedwith a blue (488-nm) and red diode (635-nm) laser. Monoclonal antibody reagents for identification andenumeration of mature human lymphocyte subsets in peripheralblood of normal individuals and patients with certain immunedysfunction. MultiTEST CD3/CD16+56/CD45/CD19 provides percentagesand absolute counts of T (CD3+), NK (CD3-CD16+CD56) andB (CD3-CD19+) lymphocytes.
Trucount Tubes
IntendedUse/Indicationsfor Use	BD Trucount tubes are intended for use with appropriated BD IVDreagent products on BD FACSLyric, BD FACSVia, BDFACSCanto II, BD FACSCanto and BD FACSCalibur flowcytometers to determine absolute counts of leucocytes inerythrocyte-lysed whole blood.	Intended UseTruCount Absolute Count Tubes are intended for use as an accessoryto TriTEST in vitro diagnostics, such as that described in K965053(CD3/CD4/CD45), to allow computation of positive cells per knowvolume of blood, using flow cytometry.Indications for UseFor use with FACS Loader For use with in vitro diagnostic immunophenotypingreagents For use in erythrocyte lysed whole blood For use to obtain absolute counts by flow cytometry
Multi-Check Controls
Feature/Attribute	Subject Device	Predicate Device
Intended Use/Indications for Use	BD Multi-Check Control is intended as a complete process controlfor immunophenotyping by flow cytometry. It is a control forantibody staining, red blood cell (RBC) lysis, instrument setup andperformance, and data analysis on BD FACSLyric, BDFACSCanto II, BD FACSCanto, and BD FACSCalibur flowcytometers.	Whole Blood Flow Control (WBFC) is a stabilized preparation ofhuman peripheral leukocytes and erythrocytes to be used as a controlin the complete immunophenotyping process which includes:antibody staining, RBC lysis, instrument set-up and instrumentperformance.
Multi-Check CD4 Low Control
	BD Multi-Check CD4 Low Control is intended as a completeprocess control for immunophenotyping by flow cytometry. It is acontrol for antibody staining, red blood cell (RBC) lysis, instrumentsetup and performance, and data analysis on BD FACSLyric, BDFACSCanto II, BD FACSCanto, and BD FACSCalibur flowcytometers.	StatusFlowLo is intended as a complete process control forimmunophenotyping by flow cytometry. It is a control for antibodystaining, RBC lysis, instrument set-up, instrument performance anddata analysis.

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A summary of the comparison is included as follows:

The subject device is similar to the predicate device as follows:

Used for identification and enumeration of human lymphocyte cell subsets ●
Performs the same flow cytometric assays with the same Multitest reagents and the same process controls .
Uses the same Trucount Tubes with a known number of beads per sample volume to calculate absolute cell counts .
Based on multicolor fluorescence ●
Uses red, blue and violet lasers to provide excitation illumination for the specific wavelengths of the fluorophores, only the . red and blue are used in the Multitest assays.
Uses dichroic mirrors to split fluorescence emissions into wavelength ranges for each fluorophore to be detected .
Uses photomultiplier tubes (PMTs) for the detection of fluorescence emission signal .

The subject device differs from the predicate device as follows:

Is available in four optical configurations: 3-1, 4-2, 4-2-2, and 4-3-3, whereas the Predicate in 3 optical . configurations: 4-2, 4-2-2 and 5-3.
Has a high powered red laser that yields higher detection sensitivity compared to the predicate device .
Instrument OC is performed using CS&T Beads and fluorescence compensation is established by using FC beads 7-color . kit. whereas for the predicate device. FACS 7-color setup beads are used to adjust fluorescent detector voltages, to set fluorescence compensation, and to monitor daily instrument performance. The subject device measures more parameters and is capable of more accurately assessing the instrument performance.
. Uses the optional FACS Universal Loading tubes in 30- or 40-tube rack, whereas the predicate device uses FACS Loader which accommodates up to 40 tubes in a carousel.
Smaller footprint with no separate wet for fluidics and generates much lower noise output than the predicate device. .

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5.7 Performance Data

The following bench (Table 5-5) and clinical (Table 5-6) performance data were provided to support the substantial equivalency determination.

Studv	Standard/Reference	Objective	Results
Intra-instrumentOpticalConfigurationEquivalency	N/A	To demonstrate system level performanceequivalency across the 3-1 (4-color), 4-2 (6-color), 4-2-2 (8-color), and 4-3-3 (10-color)optical configurations of the FACSLyric flowcytometer as measured by QC and setupperformance parameters generated utilizingCS&T beads and FC beads.	System level optical equivalency wasdemonstrated between the four configurations(3-1, 4-2, 4-2-2 and 4-3-3) of the FACSLyricsystem.
Inter-instrumentOpticalConfigurationEquivalency	CLSI EP09-A3,Measurement ProcedureComparison and BiasEstimation Using PatientSamples: ApprovedGuideline- Third Edition	To demonstrate performance equivalencybetween the 3-1 (4-color), 4-2 (6-color), 4-2-2(8-color), and 4-3-3 (10-color) opticalconfigurations of the FACSLyric flowcytometer through testing multiple instrumentsof different configurations (inter-instrument)using patient and normal donor samplesstained with Multitest IMK kit and Multitest6-Color TBNK in Trucount tubes.	The performance equivalency of the 6 IVDchannels between the 3-1, 4-2, 4-2-2 and 4-3-3optical configurations of the FACSLyric flowcytometer have been demonstrated.
Study	Standard/Reference	Objective	Results
MethodComparison	CLSI EP09-A3,Measurement ProcedureComparison and BiasEstimation Using PatientSamples; ApprovedGuideline- Third Edition	To determine the method bias between the●FACSLyric system with Multitestreagents and the FACSCanto II systemwith Multitest reagents on thedetermination of CD3+, CD3+CD4+,CD3+CD8+, CD19+ and CD16+56+percentage and absolute counts.To demonstrate equivalency between●FACSLyric Universal Loader (UL)acquisition and FACSLyric manualacquisition when using the Multitest IMKkit and Multitest 6-Color TBNK withTrucount tubes in the determination ofCD3+, CD3+CD4+, CD3+CD8+, CD19+and CD16+56+ percentage and absolutecounts.	The method bias between the FACSLyric andthe FACSCanto II, in the determination ofCD3+, CD3+CD4+, CD3+CD8+, CD19+ andCD16+56+ percentage and absolute counts.met the acceptance criteria using the MultitestIMK kit and Multitest 6-Color TBNK withTrucount tubes. In addition, equivalencybetween the FACSLyric UL acquisition andManual acquisition was demonstrated.
Within-sitePrecision	CLSI EP05-A3, Evaluationof Precision of QuantitativeMeasurement Procedures:Approved Guideline-ThirdEdition.	To evaluate the within-site precisionperformance of FACSLyric system using theMultitest IMK kit and Multitest 6-ColorTBNK with Trucount tubes.	All acceptance criteria were met. The within-site precision performance of FACSLyricsystem using the Multitest IMK kit andMultitest 6-Color TBNK with Trucount tubessatisfied the within-run and total precisionperformance requirements.
Whole BloodRepeatability	CLSI EP05-A3, Evaluationof Precision of QuantitativeMeasurement Procedures;Approved Guideline-ThirdEdition.	To verify repeatability performance across allthe FACSLyric instruments of differentconfigurations, using whole blood from patientand normal donor samples, stained with theMultitest IMK kit and Multitest 6-ColorTBNK with Trucount tubes.	Repeatability performance was demonstratedacross all the instruments with differentconfigurations. All lymphocyte subsetsmet %CV and SD acceptance criteria.
Linearity	CLSI EP6-A, Evaluation ofthe Linearity of QuantitativeMeasurement Procedures: AStatistical Approach:Approved Guideline.	To evaluate the linear range of the FACSLyricsystem using the Multitest IMK kit andMultitest 6-Color TBNK with Trucount tubes.	Acceptable linear ranges have beenestablished for each lymphocyte subset foreach of the Multitest reagents on FACSLyricsvstem.
Study	Standard/Reference	Objective	Results
Sample Carryover	CLSI H26-A2, Validation,Verification, and QualityAssurance of AutomatedHematology Analyzers;Approved Standard- SecondEdition	To determine the percent sample carryoverbased on acquisition of three highlyconcentrated samples immediately followedby acquisition of three low concentratedsamples on FACSLyric system.	All the results met the acceptance criteria ofsystem carryover less than or equal to 0.1%for low carryover and less than or equal to0.5% for standard carryover on all threeFACSLyric instruments. To meet the lowcarryover requirement the resultsdemonstrated that only a single SIT flush wasrequired.
Reagent Carryover	CLSI H26-A2 Validation,Verification, and QualityAssurance of AutomatedHematology Analyzers;Approved Standard- SecondEdition.	To verify the capability of the FACSLyricsystem to acquire samples with minimalreagent carryover.	Results met the acceptance criteria were met.The amount of reagent being carried over isfound to be well below the amount of reagentrequired for effective staining of the sample.
DetectionCapability of LoBand LoD	CLSI EP17-A2, Evaluationof Detection Capability forClinical LaboratoryMeasurement Procedures;Approved Guideline-SecondEdition.	To evaluate the limit of blank (LoB) and limitof detection (LoD) of the FACSLyric systemwith Multitest IMK kit and Multitest 6-ColorTBNK with Trucount tubes.	The detection capability parameters for thelimit of blank (LoB) and limit of detection(LoD) of the FACSLyric system wereestablished for BD Multitest reagents. ForCD4 lymphocyte subset absolute counts ofeach of the reagents, the established LoB andLoD were less than 50 cells/µL.
DetectionCapability of LoQ	CLSI EP17-A2, Evaluationof Detection Capability forClinical LaboratoryMeasurement Procedures;Approved Guideline-SecondEdition.	To evaluate the limit of quantitation (LoQ) ofthe FACSLyric system using Multitest IMKkit and Multitest 6-Color TBNK, both withTrucount tubes.	The LoQ of the FACSLyric system withMultitest reagents was established for theCD3+, CD3+CD4+, CD3+CD8+, CD19+,CD16+CD56+ and CD45+ lymphocytesubsets. The acceptance criteria were met andthe LOQ is less than 50 cells/µL for CD4lymphocyte subset absolute counts.
Setup and QCPerformance	N/A	To verify that Setup & QC Performance of theFACSLyric system meets the pre-definedspecifications.	All the Setup & QC performance results of theFACSLyric system met the acceptance criteria
Study	Standard/Reference	Objective	Results
Setup and QC Stability	N/A	To verify the stability of system level Setup & QC on the FACSLyric system.	All the system level Setup & QC stability performance results on the FACSLyric system met the pre-defined specifications.
Study	Standard/Reference	Testing Approach	Results
MethodComparison	CLSI EP09-A3,Measurement ProcedureComparison and BiasEstimation Using PatientSamples; ApprovedGuideline- Third Edition	To evaluate performance equivalence of theFACSLyric system and the FACSCanto IIsystem on the determination of lymphocytesubsets using the Multitest 6-Color TBNK andMultitest IMK kit, both with Trucount tubes,using remnant, de-linked patient specimens.	A total of 332 specimens were enrolled fortesting with Multitest 6-Color TBNK (297evaluable), and 368 specimens were enrolled forMultitest IMK kit (336 evaluable) at 5 clinicalsites. The acceptance criteria for all lymphocytesubsets were met for the FACSLyric system.The differences of the results between all thespecimens and the non-manipulated specimenswere very minimal, and no bias trend wasobserved. Additional analysis per site and bydifferent EDTA anticoagulant formulations(EDTA-K2 vs EDTA-K3) in the bloodcollection tubes also showed minimal differencewith no trend observed.
Inter-laboratoryReproducibility	CLSI EP05-A3,Evaluation of Precision ofQuantitative MeasurementProcedures; ApprovedGuideline-Third Edition.	To evaluate inter-laboratory reproducibility forthe FACSLyric system on the determination oflymphocyte subsets using the Multitest 6-Color TBNK and Multitest IMK kit inTrucount tubes with a single lot of Streck CD-Chex Plus process control material.	Inter-laboratory reproducibility was carried outat four clinical sites. The variability for withinrun, between run, between day, between site andtotal precision per reagent and lymphocytesubset was evaluated. The 97.5% one-sidedconfidence interval (CI) on the within run andtotal precision was estimated and compared to
Reference Intervals	CLSI EP28-A3c How toDefine and DetermineReference Intervals in theClinical Laboratory,Approved Guideline-Second Edition.	To re-establish the reference intervals of anormal male and female adult cohort, free ofhematological abnormality, using theFACSLyric system for the Multitest 6-ColorTBNK and Multitest IMK kit, both withTrucount tubes, on determination oflymphocyte subsets using prospectivelyprocured and de-linked donor specimens atone study site.	A total of 136 subjects were enrolled (134evaluable for Multitest 6-Color TBNK and 130evaluable for Multitest IMK kit) from oneclinical site. Reference Intervals for alllymphocyte subsets in the Multitest 6-ColorTBNK and Multitest IMK kit were establishedfor the FACSLyric system. Differences wereobserved between genders, but these differencesare not clinically significant.
Study	Standard/Reference	Testing Approach	Results
Specimen Stability	N/A	To generate data to support stability of venouswhole blood with EDTA anticoagulant whentesting the Multitest 6-Color TBNK andMultitest IMK kit, both with Trucount tubes,on the FACSLyric system using prospectivelyprocured and de-linked patient specimensfrom at least two study sites.	A total of 227 specimens were enrolled for eachof the reagents from the two clinical sites. 180specimens were evaluable for Multitest 6-ColorTBNK and 186 were evaluable for MultitestIMK kit. The results from the study support theclaim of up to 24 hours Age of Blood and 6hours Age of Stain for Multitest 6-Color TBNK.and up to 48 hours Age of Blood and 24 hoursAge of Stain for Multitest IMK kit (includeMultitest CD3/CD8/CD45/CD4 and MultitestCD3/CD16+CD56/CD45/CD19) on FACSLyricsystem.

Table 5-5 Bench Performance Summary

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Table 5-6 Clinical Performance Summary

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5.8 Conclusion

The FACSLyric flow cytometer demonstrates substantial equivalency to the FACSCanto II system with Multitest reagents.

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.”