CMV BRITE TURBO KIT

K991650 · Biotest Diagnostics Corp. · LIN · Jul 12, 1999 · Microbiology

Device Facts

• Record ID: K991650
• Device Name: CMV BRITE TURBO KIT
• Applicant: Biotest Diagnostics Corp.
• Product Code: LIN · Microbiology
• Decision Date: Jul 12, 1999
• Decision: SESE
• Submission Type: Special
• Regulation: 21 CFR 866.3175
• Device Class: Class 2

Intended Use

The CMV Brite™ Turbo Kit is intended for the qualitative detection of Cytomegalovirus (CMV) lower matrix protein pp65 by indirect immunofluorescence using microscopy in isolated peripheral blood leukocytes obtained from ethylenediaminetetraacetic acid (EDTA) or heparin anticoagulated human peripheral blood. The detection of CMV pp65 in human perpheral blood cells aids in the diagnosis of acute or reactivated CMV infection. This product is not FDA cleared (approved) for use in testing (i.e., screening) of blood or plasma donors.

Device Story

Assay detects CMV lower matrix protein pp65 in peripheral blood leukocytes; utilizes cocktail of two monoclonal antibodies (C10/C11). Process: direct lysis of erythrocytes; cytocentrifugation of leukocytes onto slides; fixation and permeabilization; indirect immunofluorescence staining with FITC-labeled secondary antibody. Performed in clinical laboratory setting by trained personnel. Clinician views slides via fluorescence microscope; CMV antigen-positive cells exhibit homogeneous yellow-green polylobate nuclear staining. Quantitative output involves counting antigen-positive cells per duplicate stain. Rapid 2-hour procedure aids diagnosis of acute or reactivated CMV infection.

Clinical Evidence

Bench testing only. Performance characteristics compared to predicate device (K951550). Sensitivity reported as 87.4% (vs 91.0% for predicate); specificity 98.8% (vs 99.5% for predicate). Positive predictive value 92.4% and negative predictive value 97.9%.

Technological Characteristics

In vitro diagnostic reagent kit. Uses monoclonal antibody cocktail (C10/C11) for indirect immunofluorescence. Requires fluorescence microscope. Specimen preparation involves direct erythrocyte lysis and cytocentrifugation. No electronic or software components; manual assay.

Indications for Use

Indicated for qualitative detection of CMV pp65 protein in isolated peripheral blood leukocytes from EDTA or heparin anticoagulated human blood to aid in diagnosis of acute or reactivated CMV infection. Not for blood or plasma donor screening.

Regulatory Classification

Identification

Cytomegalovirus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to cytomegalovirus in serum. The identification aids in the diagnosis of diseases caused by cytomegaloviruses (principally cytomegalic inclusion disease) and provides epidemiological information on these diseases. Cytomegalic inclusion disease is a generalized infection of infants and is caused by intrauterine or early postnatal infection with the virus. The disease may cause severe congenital abnormalities, such as microcephaly (abnormal smallness of the head), motor disability, and mental retardation. Cytomegalovirus infection has also been associated with acquired hemolytic anemia, acute and chronic hepatitis, and an infectious mononucleosis-like syndrome.

Predicate Devices

• CMV Brite™ Test Kit (K951550)

Submission Summary (Full Text)

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991650

### 510(k) SUMMARY OF INFORMATION RESPECTING SAFETY AND 11. EFFECTIVENESS

.

- Name and Address of Submitter A.

| | Company Name and Address: | Biotest Diagnostics Corporation<br>66 Ford Road, Suite 131<br>Denville, NJ 07834 |
|----|-------------------------------|----------------------------------------------------------------------------------|
| | Telephone: | 609-397-8511 |
| | FAX: | 609-397-8224 |
| | Contact Person: | Patricia E. Bonness |
| B. | Device Names | |
| | Proprietary Name: | CMV Brite™ Turbo Kit |
| | Common Name: | Cytomegalovirus Antigen Detection |
| | Classification Name: | Cytomegalovirus serological reagents |
| C. | Legally Marketed Device | |
| | CMV Brite™ Test Kit (K951550) | |

1 2

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## D. Device Description

The CMV antigenemia assay has been developed using a cocktail of two monoclonal antibodies (C10/C11) directed against CMV lower matrix protein pp65(6). The assay uses the C10/C11 cocktail in an indirect immunofluorescence staining of cytospin preparations of peripheral blood leukocytes. The CMV Brite™ Turbo antigenemia assay is completed within two hours of blood collection which saves time and means a rapid answer for the clinician. The CMV Brite™ Turbo method consists of:

- Direct lysis of peripheral blood erythrocytes a.
- Preparation of cytospin slides b.
- Fixation and permeabilization C.
- Indirect immunofluorescence staining using monoclonal antibodies directed d. against CMV pp65 protein
- Reading and evaluation of results e.

The first step in the CMV Brite™ Turbo method involves direct lysis of the peripheral blood erythrocytes(22). Following lysis the leukocytes are cytocentrifuged onto a slide, fixed and permeabilized to allow subsequent detection of CMV pp65 antigen. The presence of the CMV pp65 antigen is detected by the C10/C11 antibody cocktail and visualized by means of a specific secondary FITC-labeled antibody. CMV antigenpositive leukocytes exhibit homogeneous yellow-green polylobate nuclear staining when observed using a fluorescence microscope. The number of CMV antigen-positive cells are counted per duplicate stain.

The whole procedure can be performed in approximately 2 hours. The total analysis time has been shortened by performing direct erythrocyte lysis on whole blood and avoiding dextran sedimentation. Further time has been saved by shortening individual steps in the protocol so that the whole CMV antigenemia procedure has been reduced in time by more than 50%.

- Intended Use E.
The CMV Brite™ Turbo Kit is intended for the qualitative detection of Cytomegalovirus (CMV) lower matrix protein pp65 by indirect immunofluorescence using microscopy in isolated peripheral blood leukocytes obtained from ethylenediaminetetraacetic acid (EDTA) or heparin anticoagulated human peripheral blood. The detection of CMV pp65 in human perpheral blood cells aids in the diagnosis of acute or reactivated CMV infection. This product is not FDA cleared (approved) for use in testing (i.e., screening) of blood or plasma donors.

#### F. Comparison with Predicate Device

A summary comparison of the features of the CMV Brite™ Turbo Kit and CMV Brite™ Test Kit is provided in Table 1 on the following page:

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# Table 1 Feature Comparison of the CMV Brite™ and CMV Brite™ Turbo Tests

| | CMV Brite™ | CMV Brite™ Turbo |
|----------------------------------------|-------------------------------------------------------------------------|------------------------------------------------------------------------|
| Intended Use | Detection of CMV protein pp65 in<br>peripheral leukocytes | Detection of CMV protein pp65 in<br>peripheral leukocytes |
| Monoclonal Antibodies | Clones C10 and C11 | Clones C10 and C11 |
| Assay Method | Immunofluorescence Staining | Immunofluorescence Staining |
| Specimen: Type<br>Minimum vol | Heparin or EDTA<br>5 - 7 ml | Heparin or EDTA<br>3 - 5 ml |
| Preparation of<br>Leukocyte Suspension | Dextran sedimentation and lysis of red<br>cells with NH₄Cl | Direct lysis of red cells with NH₄Cl |
| | Centrifugation: 10 min at 300 xg | Centrifugation: 2 min at 1000 xg |
| Cell Count Required<br>(cells/ml) | $1.5 x 10^6$ | $2 x 10^6$ |
| Affixing Method | Cytospin Centrifugation | Cytospin Centrifugation |
| Fixation Time | 10 minutes | 5 minutes |
| Permeabilization Time | 5 minutes | 1 minute |
| Staining | 0.002% Evans Blue | 0.00075% Evans Blue |
| Incubation Time | 30 minutes | 20 minutes |
| Microscope | Immunofluorescence | Immunofluorescence |
| Controls | Positive & negative control slide (20)<br>Leukocytes & SF9 Insect cells | Positive & negative control slide (5)<br>Leukocytes & SF9 Insect cells |
| Sensitivity | 91.0% | 87.4% |
| Specificity | 99.5% | 98.8% |
| Positive Predictive<br>Value | 98.9% | 92.4% |
| Negative Predictive<br>Value | 95.7% | 97.9% |
| Kit Size | 100 test | 110 tests |
| Total Assay Time | 5 hours | 2 hours |

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## G. Performance Data

The performance characteristics of the CMV Brite™ Turbo Kit are the same as those established for the CMV Brite™ Test Kit.

- H. Conclusions Drawn from the Studies
The conclusions drawn from the design control and validation studies demonstrate that the CMV Brite™ Turbo Kit is as safe, effective, and performs as well as the legally marketed device to which equivalence is claimed, the CMV Brite™ Test Kit.

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Image /page/4/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the top half of the circle. Inside the circle is a stylized symbol that resembles a human figure in profile, with three overlapping shapes suggesting a head, body, and legs.

Food and Drug Administration 9200 Corporate Boulevard Rockville MD 20850

JUL 1.2 1999

Patricia E. Bonness Official Correspondent Biotest Diagnostics Corporation 66 Ford Road, Suite 131 Denville, New Jersey 07834

Re: K991650 Trade Name: CMV Brite™ Turbo Kit Regulatory Class: II Product Code: LIN Dated: June 28, 1999 Received: July 1, 1999

Dear Ms. Bonness:

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 Ouality 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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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 [phone].

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 [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. 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 [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours.

Steven Autman

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

Enclosure

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Page of

510(k) Number (if known): _ K 991650

Device Name: ________________________________________________________________________________________________________________________________________________________________

Indications For Use:

The CMV Brite™ Turbo Kit is intended for use in the qualitative detection of Cytomegalovirus (CMV) lower matrix protein pp65 by indirect immunofluorescence using microscopy in isolated peripheral blood leukocytes obtained from ethylenediaminetreaacetic acid (EDTA) or heparin anticoagulated human peripheral blood. The detection of CMV pp65 in human peripheral blood cells aids in the diagnosis of acute or reactivated CMV infection. This product is not FDA cleared (approved) for use in testing (i.e., screening ) of blood or plasma donors.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Woody Dubois
(Division Sign off)

Division of Clin 510(k) Number

Prescription Use OR (Per 21 CFR 801.109)

Over-The-Counter Use_

(Optional Format 1-2-96)