Predicate Devices

Premier EHEC (K953362)

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a standard immunochromatographic rapid test based on antibody-antigen binding and visual line formation. There is no mention of computational analysis, algorithms, or learning processes.

Therapeutic

No
The device is an in vitro diagnostic test used to aid in the diagnosis of diseases caused by EHEC infections by detecting Shiga toxins, not to treat or directly manage a disease or condition.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the device is "used in conjunction with the patient's clinical symptoms and other laboratory tests to aid in the diagnosis of diseases caused by enterohemorrhagic E. coli (EHEC) infections." This clearly indicates its role in the diagnostic process.

SaMD (Software as a Medical Device)

No

The device description clearly outlines a physical immunochromatographic rapid test with a sample port, test window, and control window, utilizing physical components like chromatography paper and gold-labeled antibodies. This is a hardware-based diagnostic test, not a software-only device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states that the device is for the "qualitative detection of Shiga toxins 1 and 2... in cultures derived from clinical stool specimens." It is used "in conjunction with the patient's clinical symptoms and other laboratory tests to aid in the diagnosis of diseases caused by enterohemorrhagic E. coli (EHEC) infections." This clearly indicates that the device is intended to be used in vitro (outside the body) to examine specimens (stool cultures) for diagnostic purposes.
Device Description: The description details an immunochromatographic rapid test that analyzes a sample (from a stool culture) to detect specific substances (Shiga toxins). This process is performed in vitro.
Specimen Type: The device uses "clinical stool specimens" which are biological samples taken from a patient.
Intended User: The intended users are "laboratory professionals," which is typical for IVD devices.

All of these factors align with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

ImmunoCard STAT! EHEC is an immunochromatographic rapid test for the qualitative detection of Shiga toxins 1 and 2 (also called Verotoxins) produced by E. coli in cultures derived from clinical stool specimens. ImmunoCard STAT! EHEC is used in conjunction with the patient's clinical symptoms and other laboratory tests to aid in the diagnosis of diseases caused by enterohemorrhagic E. coli (EHEC) infections.

Product codes (comma separated list FDA assigned to the subject device)

GMZ

Device Description

Each ImmunoCard STAT! EHEC test kit contains three reagents/components.

1. Test Devices, in individual foil pouches with desiccants.
1. Sample Diluent (Negative Control) in a plastic dropper vial
1. Positive Control in a plastic dropper vial
  No calibrators are needed for this device.
  This assay includes an internal control line that is used to demonstrate that the broth samples have been applied, that it has flowed correctly, and that the conjugated detector antibodies are active at the time of use. A colorless to faint pink background around the Toxin 1 and 2 Test Lines serves as a negative control and indicates the reagents were performing correctly at the time of testing.
  The Positive Control external control reagent is used with the Sample Diluent/Negative Control when external control testing is warranted. The readents also serve as indicators that the test was performed correctly, that reagents were active and specific at the time of use, and that the Test Device membrane supports proper sample flow.
  ImmunoCard STAT! EHEC is an immunochromatographic rapid test utilizing monoclonal antibodies labeled with red-colored gold particles. The test device has a circular sample port and an oval-shaped test (Toxin 1, Toxin 2) and control (Control) window.

The sample is applied to the chromatography paper via the circular sample port (Sample).
The sample is absorbed through the pad to the reaction zone containing colloidal, gold-labeled antibodies specific to Shiga toxins.
Any Shiga toxin (ST1 and ST2) antigen present complexes with the gold-labeled antibody and migrates through the pad until it encounters the binding zones in the test (Toxin 1, Toxin 2) area.
The binding zones (Toxin 1 and Toxin 2) contain another anti-ST1 or -ST2 antibody, which immobilizes any Shiga toxin-antibody comblex present. Due to the gold labeling, a distinct red line is then formed.
The remainder of the sample continues to migrate to another binding reagent zone within the control zone, and also forms a further distinct red line (positive control). Regardless of whether any Shiga toxin is present or not, a distinct red line should always be formed in the control zone and confirms that the test is working correctly.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Human stool samples

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Laboratory professionals

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

This study was conducted with samples tested fresh or following frozen storage. Samples were collected in the United States, Canada and Argentina. A total of 360 stool samples were evaluated in either GN, MacConkey (Mac) broths or both. 340 of these samples produced growth in GN broth, while 344 produced growth in Mac broth.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Clinical Trial Data / Performance Evaluation:
ImmunoCard STAT! EHEC is a modification of Duopath Verotoxin GLISA (K031367), cleared to market to detect EHEC in stool samples that had been cultured on Sorbitol MacConkey plates. This premarket submission describes the evaluations performed with clinical samples to validate the product's new intended use with broth cultures inoculated with human stool.
Samples producing discrepant results between Premier EHEC and ImmunoCard STAT! EHEC were further analyzed using cytotoxin assay. These samples generally produced weak reactions (less than 0.300) in Premier EHEC. Some of the samples grew in MAC broth but not GN broth.

Key Results from Clinical Sensitivity and Specificity (Combined Broth Total):

Sensitivity: 112/124 = 90.3% (83.7% - 94.9% CI)
Specificity: 560/560 = 100.0% (99.3% - 100% CI)
PPV: 112/112 = 100.0% (96.8% - 100% CI)
NPV: 560/572 = 97.9% (96.4% - 98.9% CI)
Correlation: 672/684 = 98.2% (97.0% - 99.1% CI)

Key Results from Clinical Sensitivity and Specificity following resolution of discrepant samples by cytotoxin assay (Combined Broth Total):

Sensitivity: 112/118 = 94.9% (89.3% - 98.1% CI)
Specificity: 566/566 = 100.0% (99.4% - 100% CI)
PPV: 112/112 = 100.0% (96.8% - 100% CI)
NPV: 566/572 = 99.0% (97.7% - 99.6% CI)
Correlation: 678/684 = 99.1% (98.1% - 99.7% CI)

Reproducibility:
Intra-assay variability was assessed with reference panel samples inoculated with ST1 and ST2. All results showed 100% precision.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

GN Broth Enrichment Cultures:

Sensitivity: 88.5% (54/61), 77.8%-95.3% CI
Specificity: 100.0% (279/279), 98.7%-100% CI
PPV: 100.0% (54/54), 93.4%-100% CI
NPV: 97.6% (279/286), 95.0%-99.0% CI
Correlation: 97.9% (333/340), 95.8%-99.2% CI

MacConkey Broth Enrichment Cultures:

Sensitivity: 92.1% (58/63), 82.4%-97.4% CI
Specificity: 100.0% (281/281), 98.7%-100% CI
PPV: 100.0% (58/58), 93.8%-100% CI
NPV: 98.3% (281/286), 96.0%-99.4% CI
Correlation: 98.5% (339/344), 96.6%-99.5% CI

Combined Broth Total:

Sensitivity: 90.3% (112/124), 83.7% - 94.9% CI
Specificity: 100.0% (560/560), 99.3% - 100% CI
PPV: 100.0% (112/112), 96.8% - 100% CI
NPV: 97.9% (560/572), 96.4% - 98.9% CI
Correlation: 98.2% (672/684), 97.0% - 99.1% CI

After Resolution of Discrepant Samples by Cytotoxin Assay:
GN Broth Enrichment Cultures:

Sensitivity: 96.4% (54/56), 87.7%-99.6% CI
Specificity: 100.0% (284/284), 98.7%-100% CI
PPV: 100.0% (54/54), 93.4%-100% CI
NPV: 99.3% (284/286), 97.5%-99.9% CI
Correlation: 99.4% (338/340), 97.9%-99.9% CI

MacConkey Broth Enrichment Cultures:

Sensitivity: 93.5% (58/62), 84.3%-98.2% CI
Specificity: 100.0% (282/282), 98.7%-100% CI
PPV: 100.0% (58/58), 93.8%-100% CI
NPV: 98.6% (282/286), 96.5%-99.6% CI
Correlation: 98.8% (340/344), 97.1%-99.7% CI

Combined Broth Total:

Sensitivity: 94.9% (112/118), 89.3% - 98.1% CI
Specificity: 100.0% (566/566), 99.4% - 100% CI
PPV: 100.0% (112/112), 96.8% - 100% CI
NPV: 99.0% (566/572), 97.7% - 99.6% CI
Correlation: 99.1% (678/684), 98.1% - 99.7% CI

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Premier EHEC (K953362)

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Duopath Verotoxin GLISA (K031367)

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.3255

Escherichia coli serological reagents.(a)
Identification. Escherichia coli serological reagents are devices that consist of antigens and antisera used in serological tests to identifyEscherichia coli from cultured isolates derived from clinical specimens. Additionally, some of these reagents consist ofEscherichia coli antisera conjugated with a fluorescent dye used to identifyEscherichia coli directly from clinical specimens or cultured isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by this bacterium belonging to the genusEscherichia, and provides epidemiological information on diseases caused by this microorganism. AlthoughEscherichia coli constitutes the greater part of the microorganisms found in the intestinal tract in humans and is usually nonpathogenic, those strains which are pathogenic may cause urinary tract infections or epidemic diarrheal disease, especially in children.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 866.9.

Summary

0

K062546

SUMMARY OF SAFETY AND EFFECTIVENESS

IDENTIFICATION INFORMATION

FEB 1 4 2007

SUBMITTERS INFORMATION

This summary of 510(k) safety and effectiveness is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.20.

SUBMITTER'S NAME AND ADDRESS: Meridian Bioscience, Inc. 3471 River Hills Drive Cincinnati, OH 45244

PHONE NUMBER: (513) 271-3700

FAX NUMBER: (513) 272-5213

CONTACT PERSON: Susan Rolih Official correspondent

DATE SUMMARY PREPARED: August 29, 2006

TRADE NAME: ImmunoCard STAT! EHEC

COMMON NAME: E coli toxins detection test

CLASSIFICATION NAME: Escherichia coli serological reagents

REGULATION: 866.3255

INTENDED USES: ImmunoCard STAT! EHEC is an immunochromatographic rapid test for the qualitative detection of Shiga toxins 1 and 2 (also called Verotoxins) produced by E. coli in cultures derived from clinical stool specimens. ImmunoCard STAT! EHEC is used in conjunction with the patient's clinical symptoms and other laboratory tests to aid in the diagnosis of diseases caused by enterohemorrhagic E. coli (EHEC) infections.

PREDICATE DEVICE: Premier EHEC (K953362)

BACKGROUND: Among the E. coli human pathogens, Shiga toxin-producing strains of E. coli have gained in importance in recent years. The group of EHEC, with their highly pathogenic serovars 0157:H7. 026, 0103, 0111, 0145, and other strains are of particular concern. Production of Shiga toxins is the most common criteria for the detection of this group of bacteria. Shiga toxins can be classified into two main categories: Shiga toxin 1 (ST1) and Shiga toxin 2 (ST2). EHEC strains may produce ST1 or ST2 only or both ST1 and ST2 simultaneously. EHEC are capable of initiating life-threatening illnesses, particularly in young children, the elderly or patients with immune deficiency. The main sources of infection are contaminated, raw or insufficiently heated foods of animal origin, eg, meat and dairy products. The reservoir for EHEC is the feces of cattle. sheep and goats. These microorganisms can enter food during the processing of meat and dairy products if hygienic conditions are inadequate. The incidence of food infection caused by Shiga toxin-producing E. coli demands reliable and rapid methods of detection. In addition to traditional culture methods, immunological techniques are becoming more useful due to their improved specificity and sensitivity. ImmunoCard STAT! EHEC is an immunological screening test based on the lateral flow principle.

1

This product was previously cleared to market as Duopath Verotoxin GLISA (K031367) to identify Shiga toxin 1 or Shiga toxin 2-producing strains of E coli isolated from colonies derived from human stools cultured on Sorbitol MacConkey agar plates. This application adds now intended use to the device (use with broth cultures inoculated with human stool) and changes the name of the product.

Type of test

Qualitative, rapid, single use, immunochromatographic assay

Specimen type

Human stool samples inoculated into enrichment media (GN or Mac broth) (This application.)

Conditions for use

ImmunoCard STAT! EHEC is designed for use by laboratory professionals under normal environmental conditions. The assay, which is stored at 2-8 C when not in use, is brought to room temperature (20-25 C) before use. Normal laboratory lighting, humidity and temperature do not affect the performance of the assay.

Contraindications

There are no contraindications associated with the use of this product.

Special instrument requirements

No instruments are used with this device.

Combination with other medical devices

No other medical devices are used in combination with this device.

2

able 1. Comparison of ImmunoCard STAT! EHEC to the predicate device and its prior design

: 1

:

Characteristics	ImmunoCard STAT! EHEC	Premier EHEC (predicate)	Duopath Verotoxin GLISA (predicate)
Device Type
Technology	Single use, rapid, lateral flow immunoassay	Microwell-based enzyme-linked immunoassay	Single use, rapid, lateral flow immunoassay
In vitro diagnostic device	Yes	Yes	Yes
Control	Includes external control reagent	Includes external control reagent	No control reagent included
Calibrator	No	No	No
Assay Features
Human factors	No special equipment	EIA-related equipment	No special equipment
Sterile device	No	No	No
Mechanical safety	Not applicable	Not applicable	Not applicable
Environmental safety	Normal medical waste	Normal medical waste	Normal medical waste
Chemical hazards	None	None	None
Radiation safety	Not applicable	Not applicable	Not applicable
Intended Use
Detection of Shiga toxins 1 and 2	Yes	Yes	Yes
Differentiation between Shiga toxins 1 and 2	Yes	No	Yes
Screening test	Yes	Yes	Yes
Diagnostic test	No	No	No
Identification test	Yes	No	Yes
Monitoring therapy	No	No	No

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| ble Samples | ImmunoCard STAT! EHEC | Premier EHEC
(predicate) | Duopath Verotoxin GLISA |
|-------------------------|---------------------------------------|------------------------------|---------------------------------------|
| Droth culture | Yes | Yes | No |
| agar culture | Yes | No | Yes |
| stool | No | Yes | No |
| s/Components | | | |
| Medium | Test Device with nitrocellulose strip | Antibody-coated microwell | Test Device with nitrocellulose strip |
| gate Reagent | In Test Device | Stand alone reagent | In Test Device |
| Diluent/Negative | Yes | Yes | Yes |
| e | | | |
| ol (external) | No | Yes | No |
| rate Reagent | No | Yes | No |
| Solution | Yes | No | Yes |
| al positive run control | Yes | No | Yes |
| al negative run control | Yes | Yes | No |
| al positive control | Yes | Yes | No |
| of
/Antibodies | | | |
| re ST1 antibodies | Murine monoclonal | Murine monoclonal | Murine monoclonal |
| re ST2 antibodies | Murine monoclonal | Murine monoclonal | Murine monoclonal |
| cor ST1 Antbodies | Murine monoclonal | Rabbit polyclonal | Murine monoclonal |
| cor ST2 Antibodies | Murine monoclonal | Rabbit polyclonal | Murine monoclonal |
| e Control | Inactivated toxin | Inactivated toxin | None |
| ison of assay steps* | None | EIA-related | None |
| oment Required | Moderately complex | Moderately complex | Moderately complex |
| I of skill required | 5 | 15 | 5 |
| y steps | Pink-red band | Yellow color | Pink-red band |
| point | | OD > 0.150 (dual wavelength) | Pos = color band Neg = no color |

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Section 5

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Table 1 continued

Performance Characteristics	ImmunoCard STAT! EHEC	Premier EHEC (predicate)	Duopath Verotoxin GLISA
Clinical Sensitivity	90.3%	100%	Not calculated
Clinical Specificity	100%	97.9%	Not calculated
Precision/Reproducibility (intra-assay)	100%	100%	100%
Linearity/reportable range	N/A	N/A	N/A
Analytical limit	ST1 = 25 ug/mL	ST1 = 7 pg/well	N/A
Detection/sensitivity	ST2 = 25 ug/mL	ST2 = 15 pg/well	ST1 and ST2 = one colony per enrichment broth
Assay cutoff	N/A	OD 0.150 (dual wavelength)	N/A
Indeterminant range	N/A	N/A	N/A

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5

DEVICE DESCRIPTION AND TECHNOLOGICAL PRINCIPLES

Construct

Each ImmunoCard STAT! EHEC test kit contains three reagents/components.

1. Test Devices, in individual foil pouches with desiccants.
1. Sample Diluent (Negative Control) in a plastic dropper vial
1. Positive Control in a plastic dropper vial

Calibrators

No calibrators are needed for this device.

Controls

This assay includes an internal control line that is used to demonstrate that the broth samples have been applied, that it has flowed correctly, and that the conjugated detector antibodies are active at the time of use. A colorless to faint pink background around the Toxin 1 and 2 Test Lines serves as a negative control and indicates the reagents were performing correctly at the time of testing.

The Positive Control external control reagent is used with the Sample Diluent/Negative Control when external control testing is warranted. The readents also serve as indicators that the test was performed correctly, that reagents were active and specific at the time of use, and that the Test Device membrane supports proper sample flow.

Failure of the internal and external controls to produce the expected results suggests the test was not performed correctly.

Technological principles

ImmunoCard STAT! EHEC is an immunochromatographic rapid test utilizing monoclonal antibodies labeled with red-colored gold particles. The test device has a circular sample port and an oval-shaped test (Toxin 1, Toxin 2) and control (Control) window.
1. The sample is applied to the chromatography paper via the circular sample port (Sample).
1. The sample is absorbed through the pad to the reaction zone containing colloidal, gold-labeled antibodies specific to Shiga toxins.
3.Any Shiga toxin (ST1 and ST2) antigen present complexes with the gold-labeled antibody and migrates through the pad until it encounters the binding zones in the test (Toxin 1, Toxin 2) area.
4.The binding zones (Toxin 1 and Toxin 2) contain another anti-ST1 or -ST2 antibody, which immobilizes any Shiga toxin-antibody comblex present. Due to the gold labeling, a distinct red line is then formed.
5.The remainder of the sample continues to migrate to another binding reagent zone within the control zone, and also forms a further distinct red line (positive control). Regardless of whether any Shiga toxin is present or not, a distinct red line should always be formed in the control zone and confirms that the test is working correctly.

CLINICAL TRIAL DATA

Performance evaluation

ImmunoCard STAT! EHEC is a modification of Duopath Verotoxin GLISA (K031367), cleared to market to detect EHEC in stool samples that had been cultured on Sorbitol MacConkey plates. This premarket submission describes the evaluations performed with clinical samples to validate the product's new intended use with broth cultures inoculated with human stool.

This study was conducted with samples tested fresh or following frozen storage. Samples were collected in the United States, Canada and Argentina. A total of 360 stool samples were evaluated in either GN, MacConkey (Mac) broths or both. 340 of these samples produced growth in GN broth, while 344 produced growth in Mac broth. Table 2 identifies the ages of the patients from whom samples were collected during the study. Table 3 provides the gender of each patient. Table 4 summarizes the sensitivity obtained with GN and Mac stool broth samples. Samples producing discrepant results between Premier EHEC and ImmunoCard

6

	Specimen Type
GN Samples	Male	Female	Not defined
Clinical site 3
Total tested	37	51	213
Mean positive reaction strength	2.0	3.1	3.5
Positive reaction range	0.5-8.0	0.5-6.0	0.5-8.0
Clinical site 4
Total tested	14	25	0
Mean positive reaction strength	1.0	All Negative	N/A
Positive reaction range	0.5-5.0	All Negative	N/A
Clinical site Totals
Total tested	51	76	213
Mean positive reaction strength	1.9	3.1	3.5
Positive reaction range	0.5-5.0	0.5-6.0	0.5-8.0
MAC Samples	Male	Female	Not defined
Clinical site 3
Total tested	40	52	213
Mean positive reaction strength	2.8	3.3	3.3
Positive reaction range	0.5-7.0	0.5-7.0	0.5-6.0
Clinical site 4
Total tested	14	25	0
Mean positive reaction strength	3.0	All Negative	N/A
Positive reaction range	3.0	All Negative	N/A
Clinical site Totals
Total tested	54	77	213
Mean positive reaction strength	2.9	3.3	3.3
Positive reaction range	0.5-7.0	0.5-7.0	0.5-6.0

Table 3 Classification of patients from whom samples were collected based on gender.

7

STAT! EHEC were further analyzed using cytotoxin assay (Table 5). These samples generally produced weak reactions (less than 0.300) in Premier EHEC (Table 6). Some of the samples grew in MAC broth but not GN broth.

Patient exclusion criteria

There are no patient exclusion criteria associated with ImmunoCard STAT! EHEC.

Influence of other disease states

There is no influence by other disease states on test results.

Table 2. Categories of patients by age from whom samples were collected for clinical studies

| Patient Age and Sample Storage GN Samples | birth to 1
month | >1 month to
2 years | >2 years to
12 years | >12 years to
21 years | >21 years | Not Defined |
|--------------------------------------------|---------------------|------------------------|-------------------------|--------------------------|-----------|--------------|
| Clinical site 3 | | | | | | |
| Total tested | 1 | 4 | 12 | 10 | 62 | 213 |
| Mean positive reaction strength | All Negative | All Negative | 3.6 | 2.6 | 2.4 | 3.5 |
| Positive reaction range | All Negative | All Negative | 1.0-5.0 | 0.5-6.0 | 0.5-6.0 | 0.5-8.0 |
| Clinical site 4 | | | | | | |
| Total tested | N/A | 1 | N/A | 2 | 35 | 0 |
| Mean positive reaction strength | N/A | All Negative | N/A | All Negative | 1.0 | N/A |
| Positive reaction range | N/A | All Negative | N/A | All Negative | 1.0 | N/A |
| Clinical site Totals | | | | | | |
| Total tested | 1 | 5 | 12 | 12 | 97 | 213 |
| Mean positive reaction strength | All Negative | All Negative | 3.6 | 2.6 | 2.3 | All Negative |
| Positive reaction range | All Negative | All Negative | 1.0-5.0 | 0.5-6.0 | 0.5-6.0 | All Negative |
| Patient Age and Sample Storage MAC Samples | birth to 1
month | >1 month to
2 years | >2 years to
12 years | >12 years to
21 years | >21 years | Not Defined |
| Clinical site 3 | | | | | | |
| Total tested | 1 | 5 | 13 | 10 | 63 | 213 |
| Mean positive reaction strength | All Negative | All Negative | 3.9 | 2.6 | 3.1 | 3.3 |
| Positive reaction range | All Negative | All Negative | 0.5-6.0 | 0.5-7.0 | 0.5-7.0 | 0.5-6.0 |
| Clinical site 4 | | | | | | |
| Total tested | N/A | 1 | N/A | 2 | 36 | 0 |
| Mean positive reaction strength | N/A | All Negative | N/A | All Negative | 3.0 | N/A |
| Positive reaction range | N/A | All Negative | N/A | All Negative | 3.0 | N/A |
| Clinical site Totals | | | | | | |
| Total tested | 1 | 6 | 13 | 12 | 99 | 213 |
| Mean positive reaction strength | All Negative | All Negative | 3.9 | 2.6 | 3.0 | 3.3 |

All Negative

0.5 - 6.0

0.5 - 7.0

Positive reaction range

0.5 · 7.0

0.5 - 6.0

8

Table 6. Analysis of samples producing discrepant results

| Site | Specimen
Serial# | Specimen Storage | ICS EHEC
Result | Premier
Absorbance | Premier Interp | CTA Result |
|------|---------------------|--------------------|--------------------|-----------------------|----------------|--------------------|
| 3 | 2831 | Cary-Blair 2-8 C | Negative | 0.223 | Positive | Negative |
| 3 | 282 | Unpreserved --70 C | Negative | 0.289 | Positive | Cytotoxin Positive |
| 3 | 281 | Unpreserved -70 C | Negative | 0.387 | Positive | Negative |
| 3 | 280 | Unpreserved -70 C | Negative | 0.151 | Positive | Negative |
| 3 | 269 | Unpreserved -70 C | Negative | 0.535 | Positive | Cytotoxin Positive |
| 3 | 266 | Unpreserved -70 C | Negative | 0.159 | Positive | Negative |
| 3 | 289 | Unpreserved -70 C | Negative | 0.280 | Positive | Negative |

GN broth-enriched samples

Mac broth-enriched samples

Site	Specimen Serial #	Specimen Storage	ICS EHEC Result	Premier Absorbance	Premier Interp	CTA Result
3	2831	Cary-Blair 2-8 C	Negative	0.314	Positive	Cytotoxin Positive
3	282	Unpreserved -70 C	Negative	0.174	Positive	Negative
3	281	Unpreserved -70 C	Negative	0.239	Positive	Cytotoxin Positive
3	280	Unpreserved -70 C	Negative	0.355	Positive	Cytotoxin Positive
3	110	Unpreserved -70 C	Negative	0.157	Positive	Negative

Legend: CTA = cytotoxin assay

9

able 4 Clinical sensitivity and specificity tabl

| GN Broth Enrichment Cultures
Premier EHEC | | | | MacConkey Broth Enrichment Cultures
Premier EHEC | | | |
|----------------------------------------------|----------|----------|-------------|-----------------------------------------------------|----------|----------|-------|
| ICS EHEC GN | Positive | Negative | Total | ICS EHEC MAC | Positive | Negative | Total |
| Positive | 54 | 0 | 54 | Positive | 58 | 0 | 58 |
| Negative | 7 | 279 | 286 | Negative | 5 | 281 | 281 |
| Total | 61 | 279 | 340 | Total | 63 | 281 | 344 |
| Sensitivity | 54/61 | 88.5% | 77.8%-95.3% | Sensitivity | 58/63 | 92.1% | 82.4% |
| Specificity | 279/279 | 100.0% | 98.7%-100% | Specificity | 281/281 | 100.0% | 98.7% |
| PPV | 54/54 | 100.0% | 93.4%-100% | PPV | 58/58 | 100.0% | 93.8% |
| NPV | 279/286 | 97.6% | 95.0%-99.0% | NPV | 281/286 | 98.3% | 96.0% |
| Correlation | 333/340 | 97.9% | 95.8%-99.2% | Correlation | 339/344 | 98.5% | 96.6% |

ombined Broth Total

Total

..............................................................................................................................................................................

	Premier EHEC
ICS EHEC
Total	Positive	Negative	Total
Positive	112	0	112
Negative	12	560	572
Total	124	560	684
Sensitivity	112/124	90.3%	83.7% - 94.9%
Specificity	560/560	100.0%	99.3% - 100%
PPV	112/112	100.0%	96.8% - 100%
NPV	560/572	97.9%	96.4% - 98.9%
Correlation	672/684	98.2%	97.0% - 99.1%

82.4% - 97.4%

286 344

58

98.7% - 100% 93.8% - 100%

96.0% - 99.4% 96.6% - 99.5%

ble 5 Test Data following resolution of discrepant samples by cytotoxin assa

GN Broth Enrichment Cultures				MacConkey Broth Enrichment
ICS EHEC GN	Premier EHEC Positive	Negative	Total	ICS EHEC MAC	Premier Positive
Positive	54	0	54	Positive	58
Negative	2	284	286	Negative	4
Total	56	284	340	Total	62
Sensitivity	54/56	96.4%	87.7%-99.6%	Sensitivity	58/62
Specificity	284/284	100.0%	98.7%-100%	Specificity	282/282
PPV	54/54	100.0%	93.4%-100%	PPV	58/58
NPV	284/286	99.3%	97.5%-99.9%	NPV	282/286
Correlation	338/340	99.4%	97.9%-99.9%	Correlation	340/344

An-an experience of the results

Total

Negative

richment Cultures
TEHEC

	Premier	EHEC
ICS EHEC
Total	Positive	Negative	Total
Positive	112	0	112
Negative	6	566	572
Total	118	566	684
Sensitivity	112/118	94.9%	89.3% - 98.1%
Specificity	566/566	100.0%	99.4% - 100%
PPV	112/112	100.0%	96.8% - 100%
NPV	566/572	99.0%	97.7% - 99.6%
Correlation	678/684	99.1%	98.1% - 99.7%

84.3% - 98.2%

93.5%

286 344

282 282

58

0

98.7% - 100% 93.8% - 100%

100.0% 100.0%

:

96.5% - 99.6%
97.1% - 99.7%

98.8%

98.6%

10

EPRODUCIBILIT

ses precion. Inte-assay wabiliy vere assessed with reference panel more nouled will 51 am ST
if to 1 same in the epoded as her bother promose on a som polition in esay prese

able 7 Results of reproducibility evaluatio

| Sample ID Qual.
Result | Site 3 | |---------------------- | Run 1 | Day 1
Run 2 | Day 2
Run 1 | Day 2
Run 2 | Day 3
Run 1 | Day 3
Run 2 | Day 1
Run 1 | Day 1
Run 2 | Day 2
Run 1 | Day 2
Run 2 | Day 3
Run 1 | Day 3
Run 2 |
| 1 HP ST1 | 2 HP ST2 | 3 Cut off LP ST1 | 4 Cut off LP ST1 | 5 Cut off LP ST2 | 6 Cut off LP ST2 | 7 Cut off HN ST1 | 8 Cut off HN ST1 | 9 Cut off HN ST2 | 10 Cut off HN ST2 | 11 LN | Average high
negative value | | Average low positive
value | | Percent Correlation | Correlation of cut off
Specimens | 100% | | 100% | | | Precision 100% | | 100% | | | Sample
| | | | | Site 4 | | | | | |
---------------|---------------------------|----------------|----------------|----------------|----------------|----------------|----------------|----------------|----------------|----------------|----------------|----------------|----------------|
| | Day 1
| 7 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 8.0 | 8.0 | 7.0 | 6.0 |
| 7 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 7.0 | 8.0 | 8.0 | 6.0 | 7.0 | 7.0 |
| 3 | 2.0 | 2.5 | 2.0 | 3.0 | 2.5 | 2.0 | 1.0 | 1.0 | 2.0 | 1.0 | 1.0 | 1.0 |
| 3 | 3.0 | 3.0 | 2.5 | 2.5 | 2.5 | 2.0 | 1.0 | 1.0 | 1.0 | 1.0 | 2.0 | 1.0 |
| 3 | 2.5 | 2.5 | 2.0 | 2.0 | 2.5 | 3.0 | 1.0 | 1.0 | 1.0 | 1.0 | 2.0 | 2.0 |
| 3 | 3.0 | 2.5 | 3.0 | 3.0 | 2.5 | 3.0 | 1.0 | 1.0 | 2.0 | 1.0 | 1.0 | 1.0 |
| 0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| 0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| 0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| 0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| 0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| 2.6 | 2.6 | 2.4 | 2.6 | 2.5 | 2.5 | 1.0 | 1.0 | 1.5 | 1.3 | 1.5 | 1.3 |
| | 100% | | | | | | 100% | | | | | |
| Site 3
| | | | Site 4
| | | |
| | Site 3
| | | | Site 4
| | | |

11

Meridian Bioscience, Inc. Cincinnati, OH

510(k) application ImmunoCard STAT! EHEC

.. . . . .

Comparison to predicate

The following table compares the outcomes of trials using ImmunoCard STAT! EHEC to the data published for the predicate device, Premier EHEC.

Table 8 Comparison of the clinical performance of ImmunoCard STAT! to Premier EHEC

| Performance Characteristics in Direct Comparison to
Clinical Status or Condition
(Combined Samples) | ImmunoCard STAT!
EHEC
Combined broths | Premier EHEC
(predicate)
Combined broths |
|-----------------------------------------------------------------------------------------------------------|---------------------------------------------|------------------------------------------------|
| Clinical Sensitivity | 90.3% | 100% |
| Clinical Specificity | 100% | 97.9% |
| Predictive Value of a Positive Test | 100% | Not done |
| Predictive Value of a Negative Test | 97.9% | Not done |
| Correlation | 98.2% | 98.1% |
| Performance characteristics | | |
| Precision/Reproducibility | 100% | 100% |
| Linearity/reportable range | N/A | N/A |
| Limit of detection | ST1 = 1.25 ng/mL
ST2 = 1.25 ng/mL | ST1 = 7 pg/well
ST2 = 15 pg/well |
| Assay cutoff | N/A | OD 0.150 (dual
wavelength |

CONCLUSIONS

IlmmunoCard STAT! EHEC can be used to detect Shiga-toxin forming strains of E. coli in human stool samples. The performance of this device for the stool broth intended use is substantially equivalent to the predicate Premier EHEC

12

DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/12/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with three tail feathers, representing the department's three main goals: protecting the health of all Americans, providing essential human services, and strengthening the nation's health and well-being. The eagle is surrounded by the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement.

Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Ms. Susan Rolih Official Correspondent Meridian Bioscience, Inc. 3474 River Hills Drive Cincinnati, OH 45244

FEB 1 4 2007

Re: K062546 Trade/Device Name: ImmunoCardSTAT! EHEC Regulation Number: 21 CFR 866.3255 Regulation Name: Escherichia coli serological reagents Regulatory Class: Class I Product Code: GMZ Dated: February 6, 2007 Received: February 6, 2007

Dear Ms. Rolih:

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 Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

13

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 information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240)276-0450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours,

Sally, artur

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

14

ImmunoCard STAT! EHEC

510(k) Notification Meridian Bioscience, Inc.

INDICATIONS FOR USE STATEMENT ImmunoCard STAT! EHEC

510(K) Number: K062544

ImmunoCard STAT! EHEC is an immunochromatographic rapid test for the qualitative detection of Shiga toxins 1 and 2 (also called Verotoxins) produced by E. coli in cultures derived from dinical stool specimens. ImmunoCard STAT! EHEC is used in conjunction with the patient's clinical symptoms and other laboratory tests to aid in the diagnosis of diseases caused by enterchemorrhagic E. coli (EHEC) infections.

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

Over-The-Counter Use (21 CFR 807 Subpart C)

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

Fuddie M. Poole
Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

576/k) KO62546