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 Description

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.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the ImmunoCyt device, based on the provided 510(k) summary:

Acceptance Criteria and Device Performance

The 510(k) summary primarily focuses on demonstrating substantial equivalence to predicate devices, rather than explicit pre-defined acceptance criteria for ImmunoCyt's performance. However, the reported sensitivity and specificity values against the predicate devices imply a comparative acceptance.

Acceptance Criteria (Implied)	Reported ImmunoCyt Performance	Predicate Device 1 (Bard BTA stat Test) Performance	Predicate Device 2 (AuraTek FDP) Performance
Sensitivity	94%	66%	67%
Specificity	50%	80%	70%

Note on "Acceptance Criteria": It's crucial to understand that for a 510(k) submission, the primary "acceptance criterion" is often substantial equivalence to a legally marketed predicate device. This means demonstrating that the new device is as safe and effective as the predicate. While ImmunoCyt's stated sensitivity is higher than the predicates, its specificity is lower. The FDA ultimately found it substantially equivalent due to other factors and its intended use in conjunction with cytology.

Study Details

2. Sample sizes and data provenance:

Test Set for Sensitivity: 87 urine samples from patients with bladder tumor recurrences confirmed by histology.
Test Set for Specificity 1 (monitored patients): 154 urine samples from patients with negative cystoscopy (monitored for bladder tumor recurrence).
Test Set for Specificity 2 (normal individuals): 170 urine samples from normal individuals (without genitourinary symptoms).
Test Set for Specificity 3 (other GU disorders): 100 urine samples from patients with various genitourinary disorders other than bladder cancer, as established by cystoscopy.
Data Provenance: Samples were collected retrospectively from 14 hospital centers in Canada.

3. Number of experts used to establish the ground truth for the test set and their qualifications:

The document does not explicitly state the number of experts or their qualifications for establishing the ground truth (histology or cystoscopy results). It only states "confirmed by histology" or "established by cystoscopy." Usually, these are performed and interpreted by pathologists and urologists, respectively, but specific numbers and experience are not provided.

4. Adjudication method for the test set:

The document does not explicitly describe an adjudication method (e.g., 2+1, 3+1). The ground truth appears to be based on clinical and pathological findings (histology, cystoscopy) as standard clinical practice rather than a consensus interpretation of the ImmunoCyt results themselves for ground truth establishment.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done:

A formal MRMC comparative effectiveness study in the sense of human readers with and without AI assistance was not conducted.
However, a reproducibility study was performed that involved multiple readers and multiple laboratories to assess within-reader, between-reader, and between-laboratory variability of the ImmunoCyt device's interpretation. This is different from a comparative effectiveness study of the device aiding human readers.
- Within-reader variability: Three readers from three different laboratories, each performing triplicate and blinded readings of one panel of three patient slides and one panel of three mock samples.
- Between-reader variability: Nine readers from three different laboratories, each performing single and blinded reading of a panel of nine mock samples slides.
- Between-laboratory variability: Three readers performing single and blinded reading of four patient slides for each level of positivity.
Effect size improvement with AI vs without AI assistance: Not applicable, as this type of MRMC study was not performed.

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

The ImmunoCyt device itself is an in vitro diagnostic immunocytofluorescence assay, which implies a laboratory technician/microscopist is reading the stained cells. It's not an "algorithm" in the typical standalone AI sense. The performance reported (sensitivity and specificity) is the standalone performance of the assay as interpreted by human observers, separate from the ground truth.
Its intended use is “in conjunction with cytology” which means it's an additive test to a human-interpreted cytology result.

7. The type of ground truth used:

Clinical Ground Truth:
- For bladder cancer recurrence confirmation: Histology.
- For absence of bladder cancer in monitored patients: Negative cystoscopy.
- For normal individuals and other genitourinary disorders: Cystoscopy (to establish presence/absence of cancer or other conditions).

8. The sample size for the training set:

The document does not explicitly mention a training set size. As ImmunoCyt is an antibody-based immunocytochemistry assay, rather than a machine learning algorithm, the concept of a "training set" for model development isn't directly applicable in the same way it would be for an AI device. The manufacturing and QC processes would be more analogous to its "training."

9. How the ground truth for the training set was established:

Not applicable as it's not an AI/ML device with a distinct "training set" in the conventional sense. The "ground truth" for developing and validating the antibodies and assay components would have been established through standard biological and chemical methods during its development.

Summary

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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 floorSainte-Foy, QuébecCanada, 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.,Subsidiary of C.R. Bard Inc.Redmond, Washington
510(k) Number:	K964151
#2 Predicate Device:	Auratek FDP
Manufacturer:	Organon Teknika, B.V.
Applicant:	Perimmune Inc.1330 Piccard Dr.Rockville, MD 20850
510(k) Number:	K970353

Premarket Notification

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Device description D.

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

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

Intendeduse	ImmunoCyt is aqualitative directimmunocytofluorescenceassay intended for use inconjunction with cytologyto increase the overallsensitivity for thedetection of tumor cellsexfoliated in the urine ofpatients previouslydiagnosed with bladdercancer.	The Bard BTA stat test isan in vitro immunoassayintended for thequalitative detection ofbladder tumor associatedantigen in urine ofpersons diagnosed withbladder cancer.	AuraTek FPD is a rapidone-step gold dyeparticle lateral flowimmunoassay indicatedfor the in vitro qualitativemeasurement offibrinogen andfibrin/fibrinogendegradation products(FDP) in human urine.
Indication	Indicated for use as anaid in the managementof bladder cancer inconjunction with urinarycytology and cystoscopy.	Indicated for use as anaid in the managementof bladder cancerpatients, in conjunctionwith cystoscopy.	To be used with standardcystoscopic examinationto aid in themanagement of patientswith a history of bladdercancer.
Format	Immunocytofluorescenceassay	Immunoassay	Immunoassay
Reagents	Antibody-based	Antibody-based	Antibody-based
SampleMatrix	Urine	Urine	Urine
Analyte	Cell-associated antigensM344, LDQ10, 19A211	A human complementfactor H-related protein	Fibrinogen andFibrin/fibrinogendegradation 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

WithinReader	MockSamples	-	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
BetweenReader	MockSamples	-	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
WithinReader	PatientSlides	-	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
BetweenLab.	PatientSlides	-	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

MockSamples	-	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%)
PatientSlides	-	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 (301) 594-4588. Additionally, for questions on the promotion and advertising of Compliance and please contact the Office of Compliance at (301) 594-4639. 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 (800) 638-2041 or (301) 443-6597 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)

Regulation Number and Section

§ 864.1860 Immunohistochemistry reagents and kits.

(a)
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.(b)
Classification of immunohistochemistry devices. (1) Class I (general controls). Except as described in paragraphs (b)(2) and (b)(3) of this section, these devices are exempt from the premarket notification requirements in part 807, subpart E of this chapter. This exemption applies to IHC's that provide the pathologist with adjunctive diagnostic information that may be incorporated into the pathologist's report, but that is not ordinarily reported to the clinician as an independent finding. These IHC's are used after the primary diagnosis of tumor (neoplasm) has been made by conventional histopathology using nonimmunologic histochemical stains, such as hematoxylin and eosin. Examples of class I IHC's are differentiation markers that are used as adjunctive tests to subclassify tumors, such as keratin.(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.
(3) Class III (premarket approval). IHC's intended for any use not described in paragraphs (b)(1) or (b)(2) of this section.
(c)
Date of PMA or notice of completion of a PDP is required. As of May 28, 1976, an approval under section 515 of the Federal Food, Drug, and Cosmetic Act is required for any device described in paragraph (b)(3) of this section before this device may be commercially distributed. See § 864.3.