BD MULTITEST 6-COLOR TBNK REAGENT WITH BD TRUCOUNT TUBES WITH MODELS 337 AND 166

K060375 · Bd Biosciences · GKZ · Jun 6, 2006 · Hematology

Device Facts

• Record ID: K060375
• Device Name: BD MULTITEST 6-COLOR TBNK REAGENT WITH BD TRUCOUNT TUBES WITH MODELS 337 AND 166
• Applicant: Bd Biosciences
• Product Code: GKZ · Hematology
• Decision Date: Jun 6, 2006
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.5220
• Device Class: Class 2

Intended Use

BD Multitest 6-Color TBNK reagent with BD Trucount tubes is intended for in vitro diagnostic use with the BD FACSCanto system to identify and determine the percentages and absolute counts of T, B, and natural killer (NK) cells as well as the CD4 and CD8 subpopulations of T cells in peripheral blood.

Device Story

Reagent kit uses fluorochrome-labeled antibodies to bind leukocyte surface antigens in whole blood samples. Processed in BD Trucount tubes containing a lyophilized pellet that releases a known number of fluorescent beads upon dissolution. Samples treated with BD FACS lysing solution to remove erythrocytes. Flow cytometer detects scatter and fluorescence signals as cells and beads pass through laser beams. Software analyzes signals to determine cell size, complexity, and fluorescence intensity. Absolute counts (cells/µL) calculated by comparing cellular events to bead events. Used in clinical laboratory settings by trained personnel. Output provides clinicians with lymphocyte subset profiles to aid in patient assessment. Benefits include standardized, automated quantification of immune cell populations.

Clinical Evidence

Bench testing only. Method comparison study (n=117) against predicate device showed high correlation (R² > 0.99 for most subsets). Precision/reproducibility evaluated over 21 days; repeatability and within-device precision reported for percentages and absolute counts. Linearity established for CD4, CD8, CD3, CD19, and CD16+CD56 subsets. Reference intervals established using 123 healthy donor samples (ages 18-65).

Technological Characteristics

Six-color direct immunofluorescence reagent (CD4 PE-Cy7, CD8 APC-Cy7, CD3 FITC, CD19 APC, CD16/CD56 PE, CD45 PerCP-Cy5.5). Uses BD Trucount tubes with lyophilized beads for absolute counting. Compatible with BD FACSCanto flow cytometer. Software-based analysis of scatter and fluorescence signals. Sodium azide (0.1%) used as preservative.

Indications for Use

Indicated for use with the BD FACSCanto flow cytometer and K3 EDTA whole blood to identify and quantify T, B, and NK cells, including CD4 and CD8 T-cell subpopulations, in peripheral blood for in vitro diagnostic use.

Regulatory Classification

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.

Special Controls

*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.”

Predicate Devices

• BD Multitest IMK kit (K980858)
• BD FACSCanto system with BD FACSCanto clinical software (K041074)

Submission Summary (Full Text)

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# 510(k) Summary

This summary of safety and effectiveness is being submitted in accordance with the requirements of the Safe Medical Devices Act of 1990 and 21 CFR 807.92.

The assigned 510(k) number is ___

K060375.

JUN -6 2006

## Submitter Information

- Submitter: BD Biosciences 2350 Qume Drive San Jose, CA 95131
- Contact: Carter Navarro Regulatory Affairs Specialist Tel: [phone] Fax: [phone] carter [e-mail]

Summary Date: February 9, 2006

### Device Name and Classification

| Trade Name: | BD Multitest 6-color TBNK reagent with<br>BD Trucount tubes |
|----------------------|-------------------------------------------------------------|
| Classification Name: | Automated Differential Cell Counter |
| Regulation Number: | 21 CFR 862.5220 |
| Product Code: | GKZ |

### Substantially Equivalent Predicate Device

BD Multitest 6-color TBNK reagent with BD Trucount tubes is substantially equivalent to BD Multitest IMK kit (K980858) when used with the BD FACSCanto system with BD FACSCanto clinical software (K041074).

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#### Device Description

The BD Multitest 6-Color TBNK reagent is a six-color direct immunofluorescence reagent for use with the BD FACSCanto system to identify and determine the percentages and absolute counts (using BD Trucount tubes) of T, B, and natural killer (NK) cells as well as the CD4 and CD8 subpopulations of T cells in peripheral blood.

When a known volume of whole blood is added to the reagent in a BD Trucount tube, the fluorochrome-labeled antibodies in the reagent bind specifically to leucocyte surface antigens. The stained samples are treated with BD FACS lysing solution to lyse erythrocytes, and the lyophilized pellet in the BD Trucount tube dissolves, releasing a known number of fluorescent beads.

During acquisition, the cells and beads travel past two laser beams and scatter the laser light. The stained cells and beads fluoresce at different intensities. These scatter and fluorescence signals, detected by the flow cytometer, provide information about each cell's size, internal complexity, and relative fluorescence intensity. During analysis, the absolute number (cells/uL) of positive cells in the sample can be determined by comparing cellular events to bead events.

#### Intended Use

BD Multitest 6-Color TBNK reagent with BD Trucount tubes is intended for in vitro diagnostic use with the BD FACSCanto system to identify and determine the percentages and absolute counts of T, B, and natural killer (NK) cells as well as the CD4 and CD8 subpopulations of T cells in peripheral blood.

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# Comparison to Predicate Device

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| Characteristic | Predicate:<br>BD Multitest IMK Kit with<br>BD Trucount Tubes (K980858)<br>and BD FACSCanto System with<br>BD FACSCanto Clinical Software<br>(K041074) | Candidate:<br>BD Multitest 6-Color TBNK<br>Reagent with BD Trucount Tubes |
|--------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | Identification and determination of<br>percentages and absolute counts of the<br>following mature human lymphocyte<br>subsets in erythrocyte-lysed whole<br>blood: T lymphocytes (CD3+,<br>CD3+CD4+, and CD3+CD8+), B<br>lymphocytes (CD3-CD19+), and NK<br>lymphocytes (CD3-CD16+CD56+). | Same. |
| Components<br>(including<br>Cluster Designation,<br>Conjugation, and<br>Clone) | BD Multitest<br>CD3/CD16+CD56/CD45/CD19<br>(1 vial, 50 tests)<br>CD3 FITC (SK7) CD16 PE (B73.1) +<br>CD56 PE (NCAM16.2) CD45 PerCP (2D1) CD19 APC (SJ25C1) Buffer with 0.1% sodium azide BD Multitest<br>CD3/CD8/CD45/CD4<br>(1 vial, 50 tests) CD3 FITC (SK7) CD8 PE (SK1) CD45 PerCP (2D1) CD4 APC (SK3) Buffer with 0.1% sodium azide BD FACS Lysing Solution<br>BD Trucount Tubes (100 tubes) | BD Multitest 6-Color TBNK Reagent<br>(1 vial, 50 tests) CD4 PE-Cy7 (SK3) CD8 APC-Cy7 (SK1) CD3 FITC (SK7) CD19 APC (SJ25C1) CD16 PE (B73.1) +<br>CD56 PE (NCAM16.2) CD45 PerCP-Cy5.5 (2D1) Buffer with 0.1% sodium<br>azide BD Trucount Tubes (50 tubes)<br>BD FACS Lysing Solution<br>(not included with reagent) |

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| Characteristic | Predicate:<br>BD Multitest IMK Kit with<br>BD Trucount Tubes (K980858 )<br>and BD FACSCanto System with<br>BD FACSCanto Clinical Software<br>(K041074) | Candidate:<br>BD Multitest 6-Color TBNK<br>Reagent with BD Trucount Tubes |
|--------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------|
| Specificity | Specificities of antibodies have been<br>verified by the International Workshop<br>on Human Leukocyte Differentiation<br>Antigens. 1,2,3,4,5,6 | Same. |
| Method to Identify<br>Populations of<br>Interest | Lyse/no-wash method using a two-tube<br>panel with four-color antibody reagents<br>(one tube stained with<br>CD3/CD16+CD56/CD45/CD19, the<br>other with CD3/CD8/CD45/CD4) to<br>identify lymphocytes with specific cell-<br>surface antigens, fluorescence<br>triggering, and CD45 vs. SSC for<br>gating. Uses fluorescent beads to<br>quantify absolute counts. | Same, except uses a one-tube panel<br>with six-color antibody reagent. |

3 Ritz J, Trinchieri G, Lanier LL. NK-cell antigens: section report. In: Schlossman SF, Bournsell L, Gilks W, et al, eds. Leucocyte Typing V: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1995;2:1367-1372.

4 Cobbold SP, Hale G, Waldmann H. Non-lineage, LFA-1 family, and leucocyte common antigens: new and previously defined clusters. In: McMichael AJ, ed. Leucocyte Typing III: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1987:788-803.

5 Bernard A, Boumsell L, Hill C. Joint report of the first international workshop on human leucocyte differentiation antigens by the investigators of the participating laboratories: T2 protocol. In: Bernard A, Boumsell L, Dausset J, Milstein C, Schlossman SF, eds. Leucocyte Typing. New York, NY: Springer-Verlag; 1984:25-60.

6 Nadler LM. B Cell/Leukemia Panel Workshop: summary and comments. In: Reinherz EL, Haynes BF, Nadler LM, Bernstein ID, eds. Leukocyte Typing II: Human B Lymphocytes. New York, NY: Springer-Verlag; 1986;2:3-43.

⁴ Haynes BF. Summary of T-cell studies performed during the Second International Workshop and Conference on Human Leukocyte Differentiation Antigens. In: Reinherz EL, Haynes BF, Nadler LM, Bernstein ID, eds. Leukocyte Typing II: Human T Lymphocytes. New York, NY: Springer-Verlag; 1986;1:3-30.

² Schmidt RE. Non-lineage/natural killer section report: new and previously defined clusters. In: Knapp W, Dörken B, Gilks WR, et al, eds. Leucocyte Typing IV: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1989:517-542.

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| Characteristic | Predicate:<br>BD Multitest IMK Kit with<br>BD Trucount Tubes (K980858)<br>and BD FACSCanto System with<br>BD FACSCanto Clinical Software<br>(K041074) | Candidate:<br>BD Multitest 6-Color TBNK<br>Reagent with BD Trucount Tubes |
|-------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Control | Recommend use of commercially<br>available whole blood control with<br>established values for subset<br>percentages and absolute counts. | Recommend use of two levels of<br>commercially available whole blood<br>controls with established values for<br>subset percentages and absolute<br>counts.<br>BD specifically recommends the use<br>of BD Multi-Check and BD Multi-<br>Check CD4 Low controls. |
| Instrument and<br>Software | BD FACSCanto flow cytometer with<br>BD FACSCanto clinical software<br>version 1.0. | Same, except uses version 2.0 of BD<br>FACSCanto clinical software. |
| System Setup | BD FACSCanto flow cytometer with<br>BD FACSCanto clinical software<br>version 1.0 and<br>BD FACS 7-color setup beads. | Same, except uses version 2.0 of BD<br>FACSCanto clinical software. |
| Sample and Stain<br>Stability | Anticoagulated blood stored at room<br>temperature (20-25°C) must be stained<br>within 48 hours of draw and then<br>analyzed within 24 hours of staining. | Anticoagulated blood stored at room<br>temperature (20-25°C) must be<br>stained within 24 hours of draw and<br>then analyzed within 6 hours of<br>staining. |
| Results | CD3+, CD3+CD4+, and CD3+CD8+ T<br>lymphocytes; CD3-CD19+ B<br>lymphocytes, and CD3-CD16+CD56+<br>NK lymphocytes expressed as<br>percentages of total lymphocytes or as<br>absolute counts (cells/\u03bcL) in whole<br>blood. | Same. |
| Sample Type | Erythrocyte-lysed whole blood,<br>collected in K3 EDTA blood collection<br>tubes. | Same. |

. ・・・・・・・・

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# Summary of Performance Data

Substantial equivalence and the performance of BD Multitest 6-color TBNK reagent with BD Trucount tubes have been demonstrated through accuracy, precision, linearity, and sample and stain stability studies.

#### Accuracy

The accuracy study design was based on NCCLS document EP9-A2, Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline – Second Edition. The predicate method was BD Multitest IMK kit (K980858) on the BD FACSCanto system with BD FACSCanto clinical software (K041074).

| Lymphocyte<br>Subset | n | Mean Bias (%)<br>(95% CI) | Range for Test System<br>(%)<br>(Predicate System) |
|----------------------|-----|---------------------------|----------------------------------------------------|
| CD4 | 117 | 0.1<br>(-0.1, 0.2) | 0.51 - 66.86<br>(0.81 - 68.67) |
| CD8 | 117 | -0.8<br>(-1.0, -0.5) | 10.77 - 83.43<br>(10.91 -83.09) |
| CD3 | 117 | - 0.2<br>(-0.4, 0.0) | 33.81 - 88.45<br>(33.12-89.51) |
| CD19 | 117 | 0.3<br>(0.2, 0.4) | 0.1 - 35.89<br>(0.08-35.77) |
| CD16+CD56 | 117 | -0.1<br>(-0.3, 0.0) | 2.44 - 51.47<br>(3.04 - 50.84) |

Lymphocyte Subset Percentages

. . . . . .

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| Lymphocyte<br>Subset | n | Mean Bias (%)<br>(95% CI) | Range for Test System (cells/µL)<br>(Predicate System) |
|----------------------|-----|---------------------------|--------------------------------------------------------|
| CD4 | 117 | -2.7<br>(-4.2, -1.3) | 4.39 - 1592.79<br>(6.23 - 1590.7) |
| CD8 | 117 | -3.5<br>(-4.4, -2.6) | 50.79 - 2416.16<br>(57.91 - 2194.02) |
| CD3 | 117 | -1.9<br>(-2.6, -1.2) | 107.14 - 3403.08<br>(108.74 - 3231.05) |
| CD19 | 117 | 1.8<br>(0.3, 3.3) | 0.5 - 1207.49<br>(0.42 - 1199.38) |
| CD16+CD56 | 117 | -2.3<br>(-4.1, 0.5) | 6.7 - 918.43<br>(7.94 - 955) |

# Lymphocyte Subset Absolute Counts

## Precision

The precision study design was based on NCCLS document EP5-A, Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline.

| Lymphocyte Subset | n | Low Sample7<br>SD | Normal Sample8<br>SD |
|-------------------|----|-------------------|----------------------|
| CD4 | 42 | 0.64 | 0.95 |
| CD8 | 42 | 1.07 | 0.65 |
| CD3 | 42 | 1.17 | 0.86 |
| CD19 | 42 | 0.89 | 0.62 |
| CD16+CD56 | 42 | 0.90 | 0.61 |

Repeatability of Lymphocyte Subset Percentages

7 Streck CD-Chex Plus CD4 Low controls.

⁸ Streck CD-Chex Plus controls.

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| Lymphocyte Subset | n | Low Sample<br>SD | Normal Sample<br>SD |
|-------------------|----|------------------|---------------------|
| CD4 | 42 | 0.69 | 1.23 |
| CD8 | 42 | 1.29 | 0.81 |
| CD3 | 42 | 1.23 | 0.90 |
| CD19 | 42 | 0.89 | 0.62 |
| CD16+CD56 | 42 | 0.96 | 0.62 |

Within-Device Precision for Lymphocyte Subset Percentages

Repeatability of Lymphocyte Subset Absolute Counts

| Lymphocyte Subset | n | Low Sample<br>%CV | Normal Sample<br>%CV |
|-------------------|----|-------------------|----------------------|
| CD4 | 42 | 7.6 | 4.7 |
| CD8 | 42 | 4.1 | 4.7 |
| CD3 | 42 | 4.0 | 4.2 |
| CD19 | 42 | 5.7 | 5.3 |
| CD16+CD56 | 42 | 7.0 | 7.9 |

Within-Device Precision of Lymphocyte Subset Absolute Counts

| Lymphocyte Subset | n | Low Sample<br>%CV | Normal Sample<br>%CV |
|-------------------|----|-------------------|----------------------|
| CD4 | 42 | 8.0 | 4.8 |
| CD8 | 42 | 5.0 | 5.4 |
| CD3 | 42 | 4.4 | 4.2 |
| CD19 | 42 | 6.0 | 5.7 |
| CD16+CD56 | 42 | 8.0 | 7.9 |

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## Linearity

The linearity study design was based on NCCLS document EP6-A, Evaluation of the Linearity of Quantitative Analytical Methods; Approved Guideline. Concentration levels were established based on an expected CD4+ range of 200 to 3,000 cells/uL.

| Lymphocyte<br>Subset | Linear Range<br>(cells/µL) | R2 |
|----------------------|----------------------------|-------|
| CD4 | 4 - 2,234 | 0.998 |
| CD8 | 158 - 1,125 | 0.991 |
| CD3 | 498 - 3,356 | 0.996 |
| CD19 | 71 - 447 | 0.989 |
| CD16+CD56 | 0 - 1,559 | 0.999 |

Lymphocyte Subset Linear Ranges for Absolute Counts

#### Sample and Stain Stability

Whole blood should be collected aseptically by venipuncture using K3 EDTA blood collection tubes. Anticoagulated blood stored at room temperature (20 – 25° C) must be stained within 24 hours of draw and then analyzed within 6 hours of staining.

# Conclusions from Performance Data

BD Multitest 6-color TBNK reagent with BD Trucount tubes is substantially equivalent to the predicate device.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/9/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is an abstract symbol resembling an eagle or bird in flight, with stylized lines forming the body and wings.

Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

JUN -6 2006

BD Biosciences c/o Mr. Carter Navarro Regulatory Affairs Specialist 2350 Qume Dr. San Jose, CA 95131-1807

Re: k060375

Trade/Device Name: BD Multitest 6-color TBNK reagent with BD Trucount tubes Regulation Number: 21 CFR 864.5220 Regulation Name: Automated Differential Cell Counter Regulatory Class: Class II Product Code: GKZ Dated: February 10, 2006 Received: February 13, 2006

Dear Mr. Navarro:

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.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), 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 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 Part 801); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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Page 2 -

This letter will allow you to begin marketing your device as described in your Section 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), please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Marie, than for
Dr. Robert Becker

Robert L. Becker, Jr., M.D., Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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# Indications for Use Statement

510(k) Number: K060375

Device Name: BD Multitest 6-color TBNK reagent with BD Trucount tubes

Indications For Use:

- For use with the BD FACSCanto flow cytometer. .
- For use with whole blood collected in K3 EDTA tubes. .
- For use in the identification and determination of percentages and absolute . counts of T, B, and natural killer (NK) cells as well as the CD4 and CD8 subpopulations of T cells in peripheral blood.
- For in vitro diagnostic use. .

X Prescription Use _ (Part 21 CFR 801 Subpart D) AND/OR

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

PLEASE DO NOT WRITE BELOW THIS LINE – CONTINUE ON ANOTHER PAGE OF NEEDED

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Maria Chan for
Josephine Bautista
DIVISION SIGN-OFF

**Office of In Vitro Diagnostic**
**Device Evaluation and Safety**

51000 K260375

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