IMMUNOCYT

K994356 · Diagnocure, Inc. · NBK · Feb 23, 2000 · Hematology

Device Facts

• Record ID: K994356
• Device Name: IMMUNOCYT
• Applicant: Diagnocure, Inc.
• Product Code: NBK · Hematology
• Decision Date: Feb 23, 2000
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.1860
• Device Class: Class 2

Intended Use

ImmunoCyt is a qualitative direct immunocytofluorescence assay intended for use in conjunction with cytology to increase the overall sensitivity for the detection of tumor cells exfoliated in the urine of patients previously diagnosed with bladder cancer. ImmunoCyt is indicated for use as an aid in the management of bladder cancer in conjunction with urinary cytology and cystoscopy.

Device Story

ImmunoCyt is an in vitro diagnostic immunocytofluorescence assay. Input: urine samples from patients with history of bladder cancer. Process: sample fixation and stabilization; application of three monoclonal antibodies (reactive to mucin and glycosylated carcinoembryonic antigen) coupled with fluorescent markers; blocking solution to minimize non-specific binding. Output: qualitative fluorescent detection of exfoliated tumor cells. Used in clinical laboratories; results interpreted by healthcare providers to supplement cytology and cystoscopy findings. Benefits: increased sensitivity for detecting recurrent bladder cancer cells compared to cytology alone.

Clinical Evidence

Clinical performance established via prospective study of 87 patients with histologically confirmed bladder tumor recurrences and 154 patients with negative cystoscopy. Sensitivity for recurrent bladder cancer was 94.3% (95% CI: 86.5-97.9%). Specificity was 50.0% (95% CI: 41.9-58.1%) in monitored patients. Additional specificity testing performed on 170 normal individuals and 100 patients with other genitourinary disorders. Reproducibility assessed via within-reader and between-reader/laboratory variability studies using mock samples and patient slides.

Technological Characteristics

In vitro diagnostic immunocytofluorescence assay. Contains three monoclonal antibodies (M344, LDQ10, 19A211) coupled with fluorescent markers. Includes sample fixative and blocking solution. Qualitative detection principle based on antibody-antigen binding. Manual assay format.

Indications for Use

Indicated for patients previously diagnosed with bladder cancer to aid in management, used in conjunction with urinary cytology and cystoscopy.

Regulatory Classification

Identification

Immunohistochemistry test systems (IHC's) are in vitro diagnostic devices consisting of polyclonal or monoclonal antibodies labeled with directions for use and performance claims, which may be packaged with ancillary reagents in kits. Their intended use is to identify, by immunological techniques, antigens in tissues or cytologic specimens. Similar devices intended for use with flow cytometry devices are not considered IHC's.

Special Controls

(2) Class II (special control, guidance document: “FDA Guidance for Submission of Immunohistochemistry Applications to the FDA,” Center for Devices and Radiologic Health, 1998). These IHC's are intended for the detection and/or measurement of certain target analytes in order to provide prognostic or predictive data that are not directly confirmed by routine histopathologic internal and external control specimens. These IHC's provide the pathologist with information that is ordinarily reported as independent diagnostic information to the ordering clinician, and the claims associated with these data are widely accepted and supported by valid scientific evidence. Examples of class II IHC's are those intended for semiquantitative measurement of an analyte, such as hormone receptors in breast cancer.

Predicate Devices

• Bard BTA stat Test (K964151)
• Auratek FDP (K970353)

Submission Summary (Full Text)

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## l. 510(k) Summary

# 510(k) number: K994356

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR §807.92.

## A. Submitter's information

| Name: | DiagnoCure Inc. |
|-----------------|------------------------------------------------------------------------------|
| Address: | 2050 René-Lévesque Ouest, 6th floor<br>Sainte-Foy, Québec<br>Canada, G1V 2K8 |
| Telephone: | 418-527-6100 |
| Fax Number: | 418-527-0240 |
| Contact person: | Pierre Pelletier |
| Date prepared: | February 7, 2000 |

## B. Device information

| Name of device: | ImmunoCyt |
|----------------------|------------------------------|
| Trade name: | ImmunoCyt™ |
| Common Name: | Immunocytofluorescence assay |
| Classification Name: | Immunohistochemistry kit |

### C. Predicate devices

ImmunoCyt is substantially equivalent to these currently marketed devices:

| #1 Predicate Device: | Bard BTA stat Test |
|----------------------|----------------------------------------------------------------------------------|
| Manufacturer: | Bainbridge Sciences Inc.,<br>Subsidiary of C.R. Bard Inc.<br>Redmond, Washington |
| 510(k) Number: | K964151 |
| #2 Predicate Device: | Auratek FDP |
| Manufacturer: | Organon Teknika, B.V. |
| Applicant: | Perimmune Inc.<br>1330 Piccard Dr.<br>Rockville, MD 20850 |
| 510(k) Number: | K970353 |

Premarket Notification

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## Device description D.

ImmunoCyt is an in vitro diagnostic device that contains a solution of three monoclonal antibodies. Two antibodies are reactive to epitopes selectively detected on a mucin found in bladder cancer cells, and one antibody reacts with a bladder cancer-associated glycosylated form of the carcinoembryonic antigen. The antibodies are coupled with fluorescent markers. This solution is used to detect tumor cells exfoliated in the urine of bladder cancer patients.

ImmunoCyt also contains a sample fixative and a blocking solution, as well as positive and negative control cells. The purpose of the blocking solution is to minimize non-specific binding of the antibodies. The purpose of the sample fixative is to optimize the quality of the cells isolated from urine by stabilizing the pH and decreasing crystals, mucus and other potential sources of artifacts in staining. Positive and negative control cells are to be used with each test sample preparation as qualitative indicators of the adequacy of the technique, reagents and instruments.

## Intended use and indications ய்

ImmunoCyt is a qualitative direct immunocytofluorescence assay intended for use in conjunction with cytology to increase the overall sensitivity for the detection of tumor cells exfoliated in the urine of patients previously diagnosed with bladder cancer. ImmunoCyt is indicated for use as an aid in the management of bladder cancer in conjunction with urinary cytology and cystoscopy.

## ட் Technological characteristics

ImmunoCyt, the in vitro diagnostic device presented in this 510(k) submission is substantially equivalent to two currently marketed devices: the AuraTek FDP and the BARD BTA stat Test. All three devices are indicated to measure the presence of bladderassociated markers in the urine of patients already diagnosed with bladder cancer, and are based upon the use of antibodies. However, the analytes detected in each system are different. All three devices are indicated for use in conjunction with cystoscopy.

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| Substantial equivalence comparison table | | | |
|------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | | | |
| Intended<br>use | ImmunoCyt is a<br>qualitative direct<br>immunocytofluorescence<br>assay intended for use in<br>conjunction with cytology<br>to increase the overall<br>sensitivity for the<br>detection of tumor cells<br>exfoliated in the urine of<br>patients previously<br>diagnosed with bladder<br>cancer. | The Bard BTA stat test is<br>an <i>in vitro</i> immunoassay<br>intended for the<br>qualitative detection of<br>bladder tumor associated<br>antigen in urine of<br>persons diagnosed with<br>bladder cancer. | AuraTek FPD is a rapid<br>one-step gold dye<br>particle lateral flow<br>immunoassay indicated<br>for the <i>in vitro</i> qualitative<br>measurement of<br>fibrinogen and<br>fibrin/fibrinogen<br>degradation products<br>(FDP) in human urine. |
| Indication | Indicated for use as an<br>aid in the management<br>of bladder cancer in<br>conjunction with urinary<br>cytology and cystoscopy. | Indicated for use as an<br>aid in the management<br>of bladder cancer<br>patients, in conjunction<br>with cystoscopy. | To be used with standard<br>cystoscopic examination<br>to aid in the<br>management of patients<br>with a history of bladder<br>cancer. |
| Format | Immunocytofluorescence<br>assay | Immunoassay | Immunoassay |
| Reagents | Antibody-based | Antibody-based | Antibody-based |
| Sample<br>Matrix | Urine | Urine | Urine |
| Analyte | Cell-associated antigens<br>M344, LDQ10, 19A211 | A human complement<br>factor H-related protein | Fibrinogen and<br>Fibrin/fibrinogen<br>degradation products |
| Sensitivity | 94% | 66% | 67% |
| Specificity | 50% | 80% | 70% |

# Substantial equivalence comparison table

The three tests are intended to detect the presence of bladder The through urine sample analysis, but they each measure a tumors through unne 'sample' analytes are all related to the presence of bladder tumors.

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#### G. Summary of studies

# Clinical performance

Samples were collected from 14 hospital centers in Canada and were mixed with an equivalent volume of ethanol and stored refrigerated until processing at DiagnoCure's laboratories in Sainte-Foy (Québec), Canada. The clinical sensitivity of ImmunoCyt was established using urine samples from a panel of 87 patients with bladder tumor recurrences confirmed by histology. The clinical specificity of ImmunoCyt was established using urine samples from a panel of 154 patients with negative cystoscopy while being followed for bladder tumor recurrence. Specificity was also tested on urine samples from a panel of 170 normal individuals (without genitourinary symptoms) and on urine samples from a panel of 100 patients with various genitourinary disorders other than bladder cancer, as established by cystoscopy.

| | 1 | 2 | 3 | Ta | T1 | T2+ | Tis | Total |
|------|------|------|------|------|------|------|------|-------|
| 1 | 21 | 45 | 21 | 52 | 23 | 7 | 5 | 87 |
| 38.1 | 38.1 | 48.9 | 66.7 | 46.2 | 43.5 | 85.7 | 80.0 | 50.6 |
| 19.0 | 19.0 | 33.9 | 43.1 | 32.5 | 23.9 | 42.0 | 29.9 | 39.7 |
| 61.3 | 61.3 | 64.0 | 84.5 | 60.4 | 65.1 | 99.2 | 98.9 | 61.4 |
| 95.2 | 95.2 | 93.3 | 95.2 | 96.2 | 95.7 | 85.7 | 80.0 | 94.3 |
| 74.1 | 74.1 | 80.7 | 74.1 | 85.7 | 76.0 | 42.0 | 29.9 | 86.5 |
| 99.8 | 99.8 | 98.3 | 99.8 | 99.3 | 99.8 | 99.2 | 98.9 | 97.9 |
| 95.2 | 95.2 | 97.8 | 100 | 96.2 | 100 | 100 | 100 | 97.7 |
| 74.1 | 74.1 | 86.8 | 86.7 | 85.7 | 87.8 | 65.2 | 54.9 | 91.2 |
| 99.8 | 99.8 | 99.9 | 100 | 99.3 | 100 | 100 | 100 | 99.6 |

## Sensitivity Results by Stage and Grade (percentage)

Kev to tables 1, and 2:

N: Number, P: Proportion; L: Lower bound of 95% confidence interva; Ly; Upper bound of 95% confidence interval

## Specificity Results by Condition (percentage)

| | N | P | L | I | C | E | S | N | I | S |
|----------------------------------------------------------|-----|------|------|------|------|------|------|------|------|------|
| Normal asymptomatic | 170 | 100 | 98.3 | 100 | 77.1 | 69.9 | 83.0 | 77.1 | 69.9 | 83.0 |
| GU disorders | | | | | | | | | | |
| Prostate cancer | 15 | 100 | 81.9 | 100 | 46.7 | 22.3 | 72.6 | 46.7 | 22.3 | 72.6 |
| Inflammation | 24 | 100 | 88.3 | 100 | 62.5 | 40.8 | 80.5 | 62.5 | 40.8 | 80.5 |
| BPH | 25 | 100 | 88.7 | 100 | 32.0 | 15.7 | 53.6 | 32.0 | 15.7 | 53.6 |
| Hematuria | 13 | 100 | 79.4 | 100 | 46.2 | 20.4 | 73.9 | 46.2 | 20.4 | 73.9 |
| Lithiasis | 8 | 100 | 68.8 | 100 | 37.5 | 10.2 | 74.1 | 37.5 | 10.2 | 74.1 |
| GU conditions* | 15 | 100 | 81.9 | 100 | 46.7 | 22.3 | 72.6 | 46.7 | 22.3 | 72.6 |
| Total | 100 | 100 | 97.0 | 100 | 46.0 | 36.1 | 56.2 | 46.0 | 36.1 | 56.2 |
| Monitored for bladder cancer with no evidence of disease | 154 | 94.8 | 89.7 | 97.6 | 50.0 | 41.9 | 58.1 | 49.4 | 41.3 | 57.5 |

Various genitourinary disorders including: vesical atony, urinational urinary disfunding, and hematuria Varrous genitous may usorders moduling. Vestair atterry, orinal y has with and hematuria.

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| otal | | | |
|-----------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----|-----|
| | | 154 | 241 |
| Negative | | | 82 |
| Positive | ದ | | 159 |
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# Comparison of ImmunoCyt and Cystoscopy in Detection of Recurrent Bladder Cancers

Monitoring sensitivity: Monitoring specificity: 94.3% (86.5-97.9%, 95% confidence interval) 50.0% (41.9-58.1%, 95% confidence interval)

#### Reproducibility 2.

Within-reader variability was determined using three readers from villnin-reader vanability was each performing triplicate and blinded three unferent laboratorise, outsing there patient slides and one panel of three reading of one panel of three-partive for each level of positivity in both panels. Between-reader variability was determined using nine parels. Between roader vanabing.
readers from three different laboratories, each reader performing readers from three different laboration of a panel of nine mock samples slides, Single and blinded foundy of positivity. Between-laboratory three slides for our vanability was dotominous as a reader performing single and blinded reading of a laboratones, odon roads. four slides for each level of positivity.

# Descriptive statistics

| Within<br>Reader | Mock<br>Samples | - | 0 | 4 | 0.44 | 1.33 | 0 | 0 | 0.00 | 0.00 |
|-------------------|-------------------|----|---|----|-------|-------|---|-----|-------|-------|
| | | + | 0 | 16 | 5.11 | 5.51 | 0 | 3 | 1.33 | 1.12 |
| | | ++ | 7 | 31 | 21.67 | 10.01 | 1 | 17 | 9.11 | 6.17 |
| Between<br>Reader | Mock<br>Samples | - | 0 | 9 | 0.41 | 1.74 | 0 | 2 | 0.19 | 0.56 |
| | | + | 0 | 18 | 5.44 | 6.62 | 0 | 9 | 1.67 | 2.27 |
| | | ++ | 7 | 81 | 35.26 | 22.74 | 3 | 65 | 14.70 | 12.22 |
| Within<br>Reader | Patient<br>Slides | - | 0 | 1 | 0.33 | 0.50 | 0 | 0 | 0.00 | 0.00 |
| | | + | 1 | 13 | 6.00 | 4.44 | 1 | 21 | 6.11 | 6.49 |
| | | ++ | 1 | 5 | 1.78 | 1.39 | 1 | 193 | 54.67 | 61.01 |
| Between<br>Lab. | Patient<br>Slides | - | 0 | 1 | 0.17 | 0.39 | 0 | 4 | 0.67 | 1.37 |
| | | + | 0 | 5 | 2.75 | 1.96 | 0 | 4 | 0.83 | 1.19 |
| | | ++ | 0 | 9 | 2.67 | 3.73 | 1 | 30 | 12.83 | 9.86 |

## Concordance rates

| Mock<br>Samples | - | Within Reader | 3/3 | 3/3 | 2/3 | 8/9 | 89% (51.8%-99.7%) |
|-------------------|--------|--------------------|-------|-------|-------|---------|-------------------|
| | | Between Reader | 8/9 | 9/9 | 6/9 | 23/27 | 85% (66.3%-95.8%) |
| | +/++ | Within Reader | 6/6 | 6/6 | 3/6 | 15/18 | 83% (58.6%-96.4%) |
| | | Between Reader | 12/18 | 18/18 | 17/18 | 47/54 | 87% (75.1%-94.6%) |
| Patient<br>Slides | - | Within Reader | 3/3 | 0/3 | 3/3 | 6/9 | 67% (29.9%-92.5%) |
| | | Between Laboratory | 3/4 | 4/4 | 0/4 | 7/12 | 58% (27.7%-84.8%) |
| | +/++ | Within Reader | 6/6 | 6/6 | 6/6 | 18/18 | 100% (81.5%-100%) |
| | | Between Laboratory | 7/8 | 8/8 | 8/8 | 23/24 | 96% (78.9%-99.9%) |
| Total | -/+/++ | All | 48/57 | 54/57 | 45/57 | 147/171 | 86% (80.8%-91.2%) |
| | | | 84.2% | 94.7% | 78.9% | | |

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## 3. Interfering substances

Testing has been conducted to identify cross-reactivity between the ImmunoCyt kit and potential contaminants in urine specimens. The specific objectives were to establish whether the presence of contaminants: 1) induced false positivity on negative samples, and 2) extinguished positivity on positive samples. Proteinaceous contaminants included white blood cells, red blood cells, albumin, and immunoglobulin. All were from human origin. Microbial contaminants included: Candida albicans, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. All microbial contaminants came from ATCC. Each test condition was conducted on three urine samples from different donors or patients, and at two pH levels (5.0 and 8.0).

| Albumin | $10 \text{ mg}$ | $10 \text{ mg}$ |
|------------------------|-------------------------------|-------------------------------|
| γ-globulin | $0.625 \text{ mg}$ | $0.625 \text{ mg}$ |
| White blood cells | $5 \times 10^4 \text{ cells}$ | $5 \times 10^4 \text{ cells}$ |
| Red blood cells | $1 \times 10^6 \text{ cells}$ | $1 \times 10^6 \text{ cells}$ |
| Candida albicans | $1 \times 10^8 \text{ cells}$ | $1 \times 10^8 \text{ cells}$ |
| Escherichia coli | $1 \times 10^8 \text{ cells}$ | $1 \times 10^8 \text{ cells}$ |
| Staphylococcus aureus | $1 \times 10^8 \text{ cells}$ | $1 \times 10^8 \text{ cells}$ |
| Pseudomonas aeruginosa | $1 \times 10^8 \text{ cells}$ | $1 \times 10^8 \text{ cells}$ |

## Interfering Substances

The effects of intravesical therapy with BCG or chemotherapy on the specificity of ImmunoCyt have not been established. Test results from samples of patients who have received such should be treated carefully.

## H. Conclusions

The safety and effectiveness data demonstrate how the performance of ImmunoCyt compares to the legally now the devices, Bard BTA stat test and AuraTek FDP. Furthermone the data substantiate the basic performance claim of the product, and to increase the overall sensitivity of urinary cytology for the detection of turnor cells exfoliated in the urine of patients previously diagnosed with bladder cancer.

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## DEPARTMENT OF HEALTH & HUMAN SERVICES

# FEB 23 2000

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

DiagnoCure, Inc. c/o Dr. Bruce F. Mackler Heller Fhrman White and McAuliffe, LLP 815 Connecticut Avenue, N.W., Suite 200 Washington, D.C. 20006-4004

Re: K994356 Trade Name: ImmunoCytTM Regulatory Class: II Product Code: NBK Dated: December 22, 1999 Received: December 23, 1999

Dear Dr. Mackler:

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 Mcdical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug, and Cosmetic Act (Act). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice. labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to moponents, are regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

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 to a legally market 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 Compliance and please contact the Office of Compliance at [phone]. Also, please note the regulation information on your responsibilities under (30 ) 34-4039. "Also, please note the regulation
information on your responsibilities under the Anton's (21CFR 807.97). Other gene information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number [phone] or [phone] or at is internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Sutman

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

Enclosure

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## Indications for Use Statement

Applicant: DiagnoCure Inc.

510(k) Number (if known): ____________________________________________________________________________________________________________________________________________________

Device Name: ImmunoCyt

Indications for use:

ImmunoCyt is a qualitative direct immunocytofluorescence assay intended for use in conjunction with cytology to increase the overall sensitivity for the detection of turnor cells exfoliated in the urine of patients previously diagnosed with bladder cancer . ImmunoCyt is indicated for use as an aid in the management of bladder cancer in conjunction with urinary cytology and cystoscopy.

Peter E. Mackin

(Division Sign-Off)
Division of Clinical Laboratory Devices K994356
510(k) Number

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

Concurrence of CDRH, Office of Device Evaluation (ODE) Prescription Use V OR Over-The-Counter Use (Per 21 C.F.R. 801.109) (Optional Format 1-2-96)