(123 days)
Premier Toxins A & B (Meridian Bioscience, Inc.), ImmunoCard Toxin A (Meridian Bioscience, Inc.), C. difficile Tox A/B II (Techlab, Inc.), Oxoid Clearview C. difficile Toxin A (Unipath, Ltd.), ColorPAC Toxin A (Becton Dickinson)
Not Found
No
The device description details a manual, visually interpreted enzyme immunoassay with no mention of computational analysis, algorithms, or data processing beyond the chemical reactions and visual readout. The performance studies describe standard clinical trial methodologies and metrics, not AI/ML model training or evaluation.
No
The device is an in vitro diagnostic (IVD) device used for diagnostic purposes to detect C. difficile toxins, not for treating any condition or disease.
Yes
The "Intended Use / Indications for Use" section explicitly states that the assay "is used as an aid in the diagnosis of C.difficile-associated disease."
No
The device description clearly outlines a physical test kit with reagents, a chromatography strip membrane, and a plastic frame. The test involves manual steps and visual reading of a color change, indicating it is a hardware-based in vitro diagnostic device, not software only.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states that the device is used "as an aid in the diagnosis of C.difficile-associated disease" by "detecting Clostridium difficile toxins A and B in human stool." This clearly indicates that the device is intended for use in vitro (outside the body) to examine specimens (human stool) to provide information for diagnostic purposes.
- Device Description: The description details a "test kit" with reagents and a "test device" designed to perform an assay on a biological sample (stool). This aligns with the nature of an in vitro diagnostic device.
- Anatomical Site: The device is used on "Human stool," which is a biological specimen collected from the human body.
- Performance Studies: The document describes clinical trials and performance metrics (sensitivity, specificity, etc.) which are standard for evaluating the performance of IVD devices.
The definition of an In Vitro Diagnostic (IVD) device is a medical device intended for use in vitro for the examination of specimens, including blood, tissue, and urine, derived from the human body, solely or principally for the purpose of providing information concerning a physiological or pathological state, or a congenital abnormality, or to determine the safety and compatibility with potential recipients, or to monitor therapeutic measures.
Based on the provided information, the ImmunoCard® Toxins A & B device fits this definition.
N/A
Intended Use / Indications for Use
ImmunoCard® Toxins A & B is a rapid, qualitative, horizontal-flow enzyme immunoassay (ElA) for detecting Clostridium difficile toxins A and B in human stool. This assay is used as an aid in the diagnosis of C.difficile-associated disease.
Product codes (comma separated list FDA assigned to the subject device)
LLH
Device Description
ImmunoCard Toxins A & B is distributed as a test kit that includes the following reagents:
ImmunoCard Toxins A & B Test Device: A chromatography strip membrane pad housed in a plastic frame and enclosed in a foil pouch with a desiccant. The membrane carries immobilized monoclonal anti-Toxin A and goat polyclonal anti-Toxin B at the TEST reaction port and crude C. difficile toxin at the CONTROL reaction port.
Sample Diluent: A buffered salt protein solution containing thimerosal (0.02%) as a preservative.
Positive Control: Inactivated crude C. difficile Toxin suspension in buffered solution containing thimerosal (0.02%) as a preservative.
Enzyme Conjugate: A blend of goat polyclonal antibodies to Toxins A and B conjugated to horseradish peroxidase and suspended in a buffered protein solution containing thimerosal (0.02%).
Wash Reagent: A buffered solution containing thimerosal (0.02%) as a preservative.
Substrate Reagent: A buffered solution containing tetramethyl-benzidine and peroxide.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
human stool
Indicated Patient Age Range
The age of the patients included in the clinical trial ranged from 1 to 99 years. Less than 2% of the 591 samples tested were from pediatric patients (≤15 years) therefore the IFU carries the limitation that the performance of specimens from pediatric patients has not been evaluated.
Intended User / Care Setting
Not Found
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
Two independent laboratories and Meridian's Development Laboratory performed testing on archival stool specimens collected from symptomatic patients that had been characterized for toxins. Each laboratory tested the samples by its own reference method (if applicable), a predicate device and ImmunoCard Toxins A and B. The status of all samples (whether positive or negative) was confirmed by cytotoxin and neutralization. Because of their expertise, Meridian's laboratory was responsible for performing all confirmation cytotoxin assays for Site 1 and Site 3.
Investigators were required to record the age and sex of the patient, the consistency, appearance and volume of each stool specimen at the time of testing. Samples were tested fresh (stored at 2-8 C for no more than 72 hours) or after frozen storage at 15 years) and Frozen (449 total samples).
Sample Type: Solid (55 samples), Semi-solid (251 samples), Liquid (285 samples).
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Clinical trials were performed at two independent laboratories and Meridian's Development Laboratory. Study design involved testing archival stool specimens from symptomatic patients using the subject device, a predicate device, and the laboratory's reference method (cytotoxin and neutralization). The total sample size was 591.
Clinical Sensitivity:
- Site 1: 41/43 (95.3%)
- Site 2: 25/28 (89.3%)
- Site 3: 34/34 (100%)
- Total Combined: 100/105 (95.2%)
Clinical Specificity:
- Site 1: 150/151 (99.3%)
- Site 2: 26/27 (96.3%)
- Site 3: 303/308 (98.3%)
- Total Combined: 479/486 (98.5%)
Predictive Value Positive Test:
- Site 1: 41/42 (97.6%)
- Site 2: 25/26 (96.2%)
- Site 3: 34/39 (87.2%)
- Total Combined: 100/107 (93.5%)
Predictive Value Negative Test:
- Site 1: 150/152 (98.5%)
- Site 2: 26/29 (89.7%)
- Site 3: 303/303 (100%)
- Total Combined: 479/484 (99.0%)
Reproducibility: 100% reproducibility/precision within and between test sites for coded specimens (low positive, moderate positive, high positive, negative, limit of detection).
Analytical Sensitivity: The limit of detection is 3 ng/mL for both toxin A and toxin B.
No high dose hook effect was observed.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Clinical sensitivity: 95.2%
- Clinical specificity: 98.5%
- Predictive value positive test: 93.5%
- Predictive value negative test: 99.0%
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 Toxins A & B (Meridian Bioscience, Inc.), ImmunoCard Toxin A (Meridian Bioscience, Inc.), C. difficile Tox A/B II (Techlab, Inc.), Oxoid Clearview C. difficile Toxin A (Unipath, Ltd.), ColorPAC Toxin A (Becton Dickinson)
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.
Not Found
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
§ 866.2660 Microorganism differentiation and identification device.
(a)
Identification. A microorganism differentiation and identification device is a device intended for medical purposes that consists of one or more components, such as differential culture media, biochemical reagents, and paper discs or paper strips impregnated with test reagents, that are usually contained in individual compartments and used to differentiate and identify selected microorganisms. The device aids in the diagnosis of disease.(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 § 866.9.
0
AUG 2 0 2004
510(k) SUMMARY OF SAFETY & EFFECTIVENESS
IDENTIFICATION INFORMATION
SUBMITTER'S INFORMATION
This summary of 510(k) safety and effectiveness is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
The assigned 510(k) number is:_K041003
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 Vice President, Regulatory Affairs and Quality Assurance Official Correspondent
DATE SUMMARY PREPARED: August 18, 2004
NAME OF DEVICE: ImmunoCard® Toxins A & B (ImmunoCard is a registered trademarks of Meridian Bioscience, Inc.)
COMMON NAME: Reagent, C. difficile toxins
CLASSIFICATION NAME: Reagents, C. difficile toxin [83(LLH)]
REGULATION: 866.2660
PREDICATE EQUIVALENT DEVICES: Premier Toxins A & B (Meridian Bioscience, Inc.), ImmunoCard Toxin A (Meridian Bioscience, Inc.), C. difficile Tox A/B II (Techlab, Inc.), Oxoid Clearview C. difficile Toxin A (Unipath, Ltd.), ColorPAC Toxin A (Becton Dickinson)
INTENDED USE:
ImmunoCard® Toxins A & B is a rapid, qualitative, horizontal-flow enzyme immunoassay (ElA) for detecting Clostridium difficile toxins A and B in human stool. This assay is used as an aid in the diagnosis of C.difficile-associated disease.
BACKGROUND:
Toxigenic Clostridium difficile is the leading cause of nosocomial infectious diarrhea in developed countries. An estimated 300,000 cases of C. difficile associated disease (CDAD) are seen per year in U.S. hospitals alone. (1.2) Clostridium difficile is the etiologic agent in approximately 25% of all cases of antibiotic-associated diarrhea. Virtually any antibiotic can predispose a patient to CDAD. The clinical presentation for CDAD ranges from asymptomatic colonization to life-threatening pseudomembranous colitis and toxic megacolon. (2) Most pathogenic strains of C. difficile produce two biologically and immunologically distinct toxin A (enterotoxin) and toxin B (cytotoxin). Toxin A was once thought to be responsible for most of the pathology seen in human CDAD until reports of clinically relevant disease caused by strains of C. difficile that produce only toxin B began to appear in the late 1990's. (1)
1
The most accurate assay overall for the detection of C. difficile is the cytotoxin assay, however the The most acourate used on the are and and is not standardized. (2) The use of memod requires lisode bakare labilition 10 && enables the physician to verify infection quickly, begin a rapid toot one and to initiate enteric isolation precautions in a hospital setting. (2)
INDICATIONS FOR USE
lmmunoCard® Toxins A & B is a rapid, qualitative, horizontal-flow enzyme immunoassay (ElA) for detecting Clostridium difficile toxins A and B in human stool. This assay is used as an aid in the diagnosis of C.difficile-associated disease.
Sypmtoms of the disease include:
- Nosocomial infectious diarrhea 1.
- Antibiotic-associated diarrhea 2.
Contraindications
There are no contraindications associated with the use of this product.
Special instrument requirements
No instruments are used with this product.
Combination with other medical devices
No other medical devices are used in combination with this device.
DEVICE DESCRIPTION AND TECHNOLOGICAL PRINCIPLES
Reagents
ImmunoCard Toxins A & B is distributed as a test kit that includes the following reagents:
ImmunoCard Toxins A & B Test Device: A chromatography strip membrane pad housed in a plastic frame and enclosed in a foil pouch with a desiccant. The membrane carries immobilized monoclonal anti-Toxin A and goat polyclonal anti-Toxin B at the TEST reaction port and crude C. difficile toxin at the CONTROL reaction port.
Sample Diluent: A buffered salt protein solution containing thimerosal (0.02%) as a preservative.
Positive Control: Inactivated crude C. difficile Toxin suspension in buffered solution containing thimerosal (0.02%) as a preservative.
Enzyme Conjugate: A blend of goat polyclonal antibodies to Toxins A and B conjugated to horseradish peroxidase and suspended in a buffered protein solution containing thimerosal (0.02%).
Wash Reagent: A buffered solution containing thimerosal (0.02%) as a preservative.
Substrate Reagent: A buffered solution containing tetramethyl-benzidine and peroxide.
Equipment needed to use the device
There is no equipment needed to use this device.
2
Cross reactivity, Interfering substances and Analytical Specificity
There are no known interfering substances that affect the performance of this device.
Drugs. Nonmicrobial Substances
Drugs, Normicrobial Substances that might be present in stool (such as fecal fat, To show that drigs of other nonmoroblar outbeatlers than may results, tests were performed with five known positive and five known negative samples spiked with the potentially interfering material. Known positive and the Rhown Hoganve Spiked with the inert agent phosphate-buffered saline (PBS), Oonlines, consisting of the barno blood of each group of spiked samples were summed and averaged then were tested in paraliti. The grades or age. A substance was considered to interfere when it changed compared to the relovant serials by +/- three or more grades from the control average. As is the Tesult of the group avorage by Illowing substances had a significant effect on the test results:
Fecal fat, metronidazole, whole blood, vancomycin, mucin, barium sulfate, Imodium AD®, Kaopectate® Caplets, Pepto Bismol®.
Microbial organisms (potentially cross-reactive species)
Microbial engaments (percents A & B is specific for C. difficile toxins, known positive and negative stool specimens were first spiked with bacterial, viral and yeast strains, then tested using ImmunoCard. The bacteria, yeast and viruses selected were those that might be expected to be present in human stools either as part of normal flora or from a disease state. The final concentration of bacteria or yeast in each sample was > 1 x 108 organisms/ml. Unspiked stool was tested in parallel to provide a reference against which the reactions with spiked stools could be compared. Reactions (the rolerse of a blue color in the test port) were graded using a 10 point scale, where "0" equals no reaction and "10" equals the strongest color development and a positive reaction. In all test cases, the Control Port was expected to produce a positive reaction of ≥2. Organisms causing interference were those that diminished positive reactions by 2 or more grades, that caused a positive to become negative, or that caused the appearance of a positive reaction in a formerly negative sample.
As is shown in Table 2, only one of the microorganisms influenced test results. The organism, a related toxigenic form of Clostridium sordellii, caused a positive result in a negative stool.
3
| Sample
ID | PBS (control) | Steric
Acid/Palmitic
Acid (fecal fat)
(4.8% w/v) | Metronidazole
(0.25% w/v) | Whole blood
(40% v/v) | Vancomycin
(0.25% w/v) | Mucin
(3.5% w/v) | Barium
sulfate
(5% w/v) | Imodium AD
Loperamide
HCl
(5% w/v) | Kaopectate
Attapulgite
(5 mg/mL)) | Pepto Bismol
(5% w/v) | | | | | |
|--------------------|---------------|-----------------------------------------------------------|------------------------------|--------------------------|---------------------------|---------------------|-------------------------------|---------------------------------------------|-----------------------------------------|--------------------------|-----|------|----|------|-----|
| | C | T | C | T | C | T | C | T | C | T | C | T | | | |
| P1 | 3 | 4 | 3 | 4 | 3 | 4/5 | 4 | 4 | 3 | 4 | 3 | 4 | 3 | 4 | |
| P2 | 3 | 3 | 3 | 2 | 4 | 4 | 3 | 3 | 2 | 3 | 4 | 3 | 3 | 3 | |
| P3 | 3 | 2 | 3 | 3 | 3 | 2 | 4 | 2 | 3 | $1/2$ | 4 | 4 | 4 | 1 | |
| P4 | 3 | 4 | 3 | 4 | 3 | 5 | 3 | 4 | $1/2$ | 4 | 3 | 4/5 | 3 | 4 | |
| P5 | 3 | 2 | 3 | $1/2$ | 3 | 2 | 3 | 2 | 4 | 2 | 4 | 3 | 2 | 2 | |
| Total +
grade | 15 | 15 | 15 | 14.5 | 16 | 17.5 | 17 | 15 | 13.5 | 14.5 | 18 | 18.5 | 15 | 15.5 | 16 |
| Average
- grade | 3 | 3 | 3 | 2.9 | 3.2 | 3.5 | 3.4 | 3 | 2.7 | 2.9 | 3.6 | 3.7 | 3 | 3.1 | 3.2 |
| N1 | 3 | 0 | 3 | 0 | 4 | 0 | 4 | 0 | 3 | 0 | 3 | 0 | 4 | 0 | |
| N2 | 3 | 0 | 4 | 0 | 4 | 0 | 4 | 0 | 3 | 0 | 4 | 0 | 2 | 0 | |
| N3 | 3 | 0 | 3 | 0 | 3 | 0 | 4 | 0 | 3 | 0 | 3 | 0 | 4 | 0 | |
| N4 | 3 | 0 | 3 | 0 | 2 | 0 | 4 | 0 | 3 | 0 | 3 | 0 | 2 | 0 | |
| N5 | 2 | 0 | 3 | 0 | 3 | 0 | $1/2$ | 0 | 4 | 0 | 3 | 0 | 3 | 0 | |
| Total
grade | 14 | 0 | 16 | 0 | 16 | 0 | 17.5 | 0 | 16 | 0 | 16 | 0 | 15 | 0 | |
| Average - grade | 2.8 | 0 | 3.2 | 0 | 3.2 | 0 | 3.5 | 0 | 3.2 | 0 | 3.2 | 0 | 3 | 0 | |
able 1. Affect of drugs and nonmicrobial substances on positive and negative test result
Control results
| | Don
Control | est Port |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------|----------|
| mmunoCard Positive Control | | |
| " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " "
'mmunoCard Negative Contrc | | |
| | | |
Legend: LP = low positive (as defined by Premier Toxins A & B ElA), C = control port, T = test por, P = positive, + = positive, - = negative
4
Table 2. Affect of microbial organisms on positive and negative test results.
| | | Spiked into
Positive Stool | | Spiked into
Negative Stool | | |
|-------------------------------------------|------------------|-------------------------------|--------------|-------------------------------|--------------|------------------------------|
| Organism | ATCC # or ID # | Control
Port | Test
Port | Control
Port | Test
Port | Organism effects
outcome? |
| | | | | | | |
| Unspiked stool control | Not applicable | 3-4 | 2-3 | 2-4 | 0 | Not applicable |
| | | | | | | |
| Adenovirus 40 (1:125) (exp. 30Jul04) | 01080032H | 3 | 2 | 4 | 0 | No |
| Adenovirus 41 (exp. 15Jan05) | 4017441.008 | 4 | 3 | 3 | 0 | No |
| Aeromonas hydrophila | 35654 | 3 | 3 | 3 | 0 | No |
| Bacillus cereus | 11778 | 3 | 2 | 3 | 0 | No |
| Bacillus subtilis | 6051 | 4 | 3 | 4 | 0 | No |
| Bacteroides fragilis | 23745 | 4 | 2 | 3 | 0 | No |
| Campylobacter coli | 49941 | 3 | 3 | 3 | 0 | No |
| Campylobacter jejuni | 29428 | 3 | 2 | 3 | 0 | No |
| Candida albicans | BHI 15004 | 3 | 3 | 4 | 0 | No |
| Clostridium butyricum | 4855 | 4 | 2 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q11 | 620-3/28 | 3 | 2 | 2 | 0 | No |
| Clostridium difficile (non-toxogenic) Q12 | UNC19904 | 2 | 2 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q13 | X-15076-590-1/20 | 3 | 3 | 4 | 0 | No |
| Clostridium difficile (non-toxogenic) Q15 | 611 | 3 | 2 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q17 | 234-25039 | 3 | 2 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q18 | 2C62-1/31 | 4 | 1 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q19 | 11186 | 3 | 2 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q20 | 2C165 | 2 | 1 | 3 | 0 | No |
| Clostridium difficile (non-toxogenic) Q32 | TCH3 | 3 | 2 | 4 | 0 | No |
| Clostridium difficile (non-toxogenic) Q31 | C124 | 2 | 2 | 4 | 0 | No |
| Clostridium perfringens | 3624 | 3 | 2 | 2 | 0 | No |
| Clostridium septicum | 12464 | 3 | 1/2 | 3 | 0 | No |
| Clostridium sordellii | VPI 9048 | 3 | 2 | 3 | 1 | Yes |
| Clostridium sordellii | 9714 | 4 | 3 | 4 | 0 | No |
| Clostridium sporogenes | 3584 | 3 | 2/3 | 4 | 0 | No |
| Enterobacter cloacae | 3251 | 3 | 2 | 4 | 0 | No |
5
Table 2 continued.
| Organism | ATCC # or ID # | Control
Port | Test
Port | Control
Port | Test
Port | Organism effects
outcome? |
|----------------------------------|-------------------------|-----------------|--------------|-----------------|--------------|------------------------------|
| Escherichia coli 0157:H7 | 43895 | 3 | 3 | 3 | 0 | No |
| Eschericia coli 0157:H7 | EMDI-012 | 3 | 2 | 3 | 0 | No |
| Escherichia coli | 8739 | 4 | 2 | 4 | 0 | No |
| Escherichia coli | 9637 | 3 | 3 | 3 | 0 | No |
| Helicobacter pylori | 43504 | 3 | 3 | 4 | 0 | No |
| Klebsiella pneumoniae | 13883 | 3 | 2 | 3 | 0 | No |
| Peptostreptococcus anaerobius | 27337 | 3 | 2 | 3 | 0 | No |
| Porphyromonas asaccharolytica | 25260 | 4 | 3 | 3 | 0 | No |
| Proteus vulgaris | 6380 | 3 | 2 | 3 | 0 | No |
| Pseudomonas aeruginosa | Meridian Microbank # 18 | 2 | 3 | 3 | 0 | No |
| Rotavirus (20Aug08) | TRV061 | 3 | 2 | 4 | 0 | No |
| Salmonella typhimurium | 14028 | 3 | 3 | 4 | 0 | No |
| Serratia liquefaciens | 35551 | 4 | 3 | 3 | 0 | No |
| Shigella dysenteriae | Meridian Microbank # 14 | 3 | 3 | 3 | 0 | No |
| Shigella flexneri | Meridian Microbank # 15 | 3 | 3 | 3 | 0 | No |
| Shigella sonnei | Meridian Microbank # 6 | 3 | 3 | 4 | 0 | No |
| Staphylococcus aureus | 6538 | 4 | 3 | 4 | 0 | No |
| Staphylococcus aureus (Cowans I) | 12598 | 4 | 4 | 4 | 0 | No |
| Staphylococcus epidermidis | 12228 | 3 | 2 | 3 | 0 | No |
| Streptococcus faecalis | Meridian Microbank # 7 | 4 | 2 | 4 | 0 | No |
| Vibrio parahaemolyticus | 17802 | 3 | 3 | 3 | 0 | No |
| Yersinia enterocolitica | Meridian Microbank # 10 | 4 | 3 | 3 | 0 | No |
Calibrators
There are no calibrators used with this device.
Controls
The assay includes an internal control (control port) that is used to demonstrate that sample has been applied, that it has flowed correctly and that the Enzyme Conjugate is active at the time of testing. In addition, a Positive Control Reagent and Sample Diluent (used for a negative control reagent) are supplied as external controls. Proper results obtained with the Control Port, Positive Control and negative control Diluent serve as indicators that the test was performed correctly, that the antibodies embedded in the membrane and the Enzyme Conjugate are active at the time of testing, and that the membrane supports proper sample flow. Failure of the internal and external control to produce the expected results suggests the test was not performed correctly (ie, incorrect volume of reagents added; incorrect incubation temperature or times used or that reagents were not brought to room temperature prior to testing).
Technological principles
ImmunoCard Toxins A & B consists of a membrane held in a plastic frame with two sample ports and two reaction ports. The membrane carries immobilized antibodies to toxins A and B. The Enzyme Conjugate Reagent consists of antibodies to toxins A and B coupled to horseradish peroxidase. To perform the test, patient stool sample is diluted with Specimen Diluent and Enzyme Conjugate and the
6
mixture is incubated for 5 minutes. During the incubation, molecules of toxin, if present, are bound to the anti-toxin antibodies of the Conjugate. Following incubation, an aliquot of the mixture is added to the anti-oally antiloodio of the Gorijagens: 's incubated for an additional 5 minutes at 20-26 C. During each of the two sample portune the tool is separated from particulate matter as the fluid the second nodbation the toxin serijagais romane to the TEST and CONTROL reaction ports. The portion of the bample frome in bug captured at the TEST reaction port by immobilized antitoxin in the reaction membrane. (The second of the two reaction ports serves as an internal control.) Both reaction ports are subsequently wash Reagent to reduce interference by contaminating roution porto and basedgement is added. The reaction ports are incubated for an additional 5 process during which time the enzyme conjugate modifies the Substrate Reagent. The result is the minutes dailing which the eneyms sen read visually. Development of a blue color in the TEST reaction port indicates a positive test. In the CONTROL port, the anti-toxin antibodies of the conjugate roution por in the immobilized toxin. The appearance of blue in the CONTROL reaction port indicates bird directly to the infribalized were active at the time of use and that proper sample migration occurred.
| Characteristics | IC Toxins A
& B | Cytotoxin/
Neutraliza
tion
(Std) | Premier
Toxins
A & B | BD
ColorPAC
Toxin A | Oxoid
C. difficile
Toxin
Detection | Wampole
C. difficile
Tox A/B II |
|-----------------------------------------------|--------------------|-------------------------------------------|----------------------------|---------------------------|---------------------------------------------|---------------------------------------|
| Device Type | | | | | | |
| In vitro diagnostic device | Yes | Yes | Yes | Yes | Yes | Yes |
| Control | No | No | No | No | No | No |
| Calibrator | No | No | No | No | No | No |
| Intended Use | | | | | | |
| Detection of Toxins A and B in human
stool | Yes | Yes | Yes | Only A | Only A | Yes |
| Detection of Toxins A and B in culture | No | Yes | Yes | Only A | No | Yes |
| Screening test | Yes | No | Yes | Yes | Yes | Yes |
| Diagnostic test | No | Yes | No | No | No | No |
| Monitoring therapy | No | Yes | No | No | No | No |
| Acceptable Sample | | | | | | |
| Formed stool | Yes | Yes | Yes | Yes | Yes | Yes |
| Semi-solid stool | Yes | Yes | Yes | Yes | Yes | Yes |
| Liquid stool | Yes | Yes | Yes | Yes | Yes | Yes |
| Stool collected in transport media | No | No | No | No | No | No |
| Broth culture | No | Yes | Yes | Yes | No | Yes |
SUBSTANTIAL EQUIVALENCE TO PREDICATE DEVICES
7
| Performance Characteristics
(rounded) in Direct Comparison
to Clinical Status or Condition | IC Toxins
A & B | Cytotoxin
(Std) | Premier
Toxins A
& B | BD
ColorPAC
Toxin A | Oxoid C.
difficile
Toxin
Detection | Wampole
C. difficile
Tox A/B II |
|--------------------------------------------------------------------------------------------------|--------------------|--------------------|----------------------------|---------------------------|------------------------------------------------|---------------------------------------|
| Clinical Sensitivity | 95.2% | | 94.7% | 81% | 83.3% | 92.2% |
| Clinical Specificity | 98.5% | | 97.3% | 97% | 96.7% | 100% |
| Predictive Value of a Positive Test
(incidence = 1%) | 93.5% | | 87.4% | ND | 91.7% | 100% |
| Predictive Value of a Negative Test | 99% | | 98.9% | ND | 93.2% | 98.6% |
| Correlation | 98% | | 96.9% | ND | ND | 98.8% |
| Laboratory Equivalence in
Stool tests with (Predicate
Device) | | | (cell
cytotoxicity) | (cytotoxin B) | (cell
cytotoxicity
and
neutralization | Tissue
culture |
| Concordance of positive tests with
predicate device | 95% | | 90/95 (95%) | 96/119 (81%) | 110/132
(83%) | 165/179
(92%) |
| Discordance of positive tests with
predicate device | 5% | | 5/95 (5%) | 23/119 (19%) | 22/132 (17%) | 14/179 (8%) |
| Concordance of negative tests with
predicate device | 98% | | 465/478
(97%) | 622/641
(90%) | 301/311
(97%) | 973/973
(100%) |
| Discordance of negative tests with
predicate device | 2% | | 13/478 (3%) | 19/641 (10%) | 10/311 (3%) | 0/973 (0%) |
| Performance characteristics | | | | | | |
| Precision/Reproducibility (intra-
assay) | 100% | | See insert | 100% | ND | ND |
| Precision/Reproducibility (inter-
assay) | 100% | | See insert | 100% | ND | ND |
| Linearity/reportable range | NA | | See insert | N/A | ND | ND |
| Limit of detection | 3 ng/mL | | 1.2 ng/mL A
2.4 ng/mL B | | ND | >0.8 ng/mL A
2.5 ng/ML B |
| Liquid stool | | | ND | 1.38-5.21 | ND | ND |
| Semi-solid stool | | | ND | 1.61 - 18.71 | ND | ND |
| Solid stool | | | ND | 3.19-22.58 | ND | ND |
| Assay cutoff | NA | | See insert | N/A | N/A | See insert |
:
8
Me.
Comparison of Assay Methods
| Characteristic | ICTAB | Premier Toxins A & B | BD ColorPAC | Oxoid C. difficile Toxin A | Wampole Tox A/B II |
|---|---|---|---|---|---|
| Intended use | Detection of toxins in human stool | Detection of toxins in human stool and broth culture | Detection of Toxin A in human stool or broth culture | Detection of Toxin A in human stool | Detection of Toxins A and B in fecal specimens and broth culture |
| Specimen Required | 1. Human stool | Human stool | 1. Human Stool |
- Broth culture | Human stool | Human stool
Human stool in prep soln |
| Technology | Horizontal-flow EIA | Microplate-based EIA | Rapid chromatographic assay | Rapid chromatographic assay | Microwell based EIA |
| Level of skill required | Moderate complexity | Moderate complexity | Moderate complexity | Moderate complexity | Moderate complexity |
| Assay steps | 1. Add 200 uL Sample Diluent to a tube - Add 3 drops Enzyme Conjugate.
- Add 25 uL stool to tube, vortex and let stand 5 min.
- Vortex sample mixture.
- Add 150 ul. diluted sample to test and control ports.
- Incubate 5 min., 20-26°C.
- Add 3 drops Wash Reagent to each port.
- Add Substrate.
- Incubate 5 min, 20-26 C.
- Read at end of incubation. | 1. Add 200 ul. Sample Diluent to a tube.
- Add 50 ul, stool and vortex.
- Add 100uL diluted sample to test well.
- Add 1 drop Enzyme Conjugate to well.
- Incubate 50 min, 35-39 C.
- Wash wells with Wash Buffer.
- Add 2 drops Substrate and shake plate and wait 30 seconds.
- At 2 drops Stop Solution.
- Wait 2 minutes then read at OD at 450 nm or 450/630nm. | 1. Add 1 mL Sample Buffer to tube.
- Add 0.5 mL or 0.5 g stool and vortex.
- Add 3 drops diluted sample to test device
- Add 1 drop Wash Reagent and absorb.
- Add 1 drop Detector A and wait 3 minutes.
- Add 1 drop Wash Reagent and absorb.
- Add 1 drop Detector B and wait 3 minutes.
- Add 1 drop Wash Reagent and absorb.
- Read after 1 minute. | 1. Add 1 mL Sample Diluent to tube.
- Add 100 uL or stool bead to tube and vortex.
- Centrifuge sample to separate solids from supernate.
- Add 125 ul supernate to Test Unit.
- Incubate 30 minutes.
- Read test at end of 30 minutes. | 1. Add 200 uL Diluent to tube.
- Add 50 uL of stool to tube and vortex.
- Add 1 drop Conjugate to test well.
- Add100 uL of diluted specimen.
- Incubate, 37 C, 50 minutes.
- Wash wells with Wash Solution
- Add 100 uL Substrate
- Incubate for 10 minutes.
- Add 1 drop Stop Solution and wait 2 minutes.
- Read at OD 450 nm. |
| End point | Appearance of blue color | Appearance of yellow color | Appearance of pink line | Appearance of blue line | Appearance of yellow color |
| Interpretation of test result | Positive = appearance of blue color in test and control ports (indicates presence of toxin)
Negative = no color in test port with blue color in reaction port (indicates absence of toxin) | Positive = appearance of yellow color in test and positive control wells (indicates presence of toxin)
Negative = no color in test well with yellow color in positive control (indicates absence of toxin) | Positive = appearance of a pink line at test and control points (indicates presence of toxin)
Negative = no color at test point with pink color at control point | Positive = appearance of a blue line at test and control points (indicates presence of toxin)
Negative = no color at test point with blue color at control point | Positive = appearance of yellow color in test and positive control wells (indicates presence of toxin)
Negative = no color in test well with yellow color in positive control (indicates absence of toxin) |
9
CLINICAL TRIALS
Two independent laboratories and Meridian's Development Laboratory performed testing on archival I wo independent laboratione and incraraneles collected from symptomatic patients that had been (it croupolited for toxing. Each laboratory tested the samples by its own reference method (if applicable), a predicate device and ImmunoCard Toxins A and B. The status of all samples (whether applicable), a products and was confirmed by cytotoxin and neutralization. Because of their positive of negative by immunoodial was evelles and the responsibility of performing all confirmation cytotoxin assays for Site 1 and Site 3.
Investigators were required to record the age and sex of the patient, the consistency, appearance and involugation thou specimen at the time of testing. Samples were tested fresh (stored at 2-8 C for no more than 72 hours) or after frozen storage at 15 Yrs | Not defined |
|---------------------------------|--------|---------|----------|----------|-------------|
| Clinical site 1 | | | | | |
| Total tested Fresh | 0 | 0 | 2 | 140 | N/A |
| Mean positive reaction strength | N/A | N/A | N/A | 6.6 | N/A |
| Positive reaction range | N/A | N/A | N/A | 1-10 | N/A |
| Total tested Frozen | 0 | 1 | 0 | 51 | N/A |
| Mean positive reaction strength | N/A | N/A | N/A | 5.7 | N/A |
| Positive reaction range | N/A | N/A | N/A | 1-10 | N/A |
| Clinical site 2 | | | | | |
| Total tested Fresh | 0 | 0 | 0 | 0 | N/A |
| Mean positive reaction strength | N/A | N/A | N/A | N/A | N/A |
| Positive reaction range | N/A | N/A | N/A | N/A | N/A |
| Total tested Frozen | 0 | 3 | 1 | 51 | N/A |
| Mean positive reaction strength | N/A | 8.0 | N/A | 8.0 | N/A |
| Positive reaction range | N/A | 1-10 | N/A | 1-10 | N/A |
| Clinical site 3 | | | | | |
| Total tested Fresh | N/A | N/A | N/A | N/A | N/A |
| Mean positive reaction strength | N/A | N/A | N/A | N/A | N/A |
| Positive reaction range | N/A | N/A | N/A | N/A | N/A |
| Total tested Frozen | N/A | N/A | N/A | N/A | 342 |
| Mean positive reaction strength | N/A | N/A | N/A | N/A | 7.1 |
| Positive reaction range | N/A | N/A | N/A | N/A | 1-10 |
| Clinical site Totals | | | | | |
| Total tested Fresh | 0 | 1 | 2 | 140 | N/A |
| Mean positive reaction strength | N/A | N/A | N/A | 7.0 | N/A |
| Positive reaction range | N/A | N/A | N/A | 1-10 | N/A |
| Total tested Frozen | 0 | 3 | 1 | 102 | 342 |
| Mean positive reaction strength | N/A | 8.0 | N/A | 8.0 | 7.1 |
| Positive reaction range | N/A | 1-10 | N/A | 1-10 | 1-10 |
| Table 3. Patient age, sample storage statistics and mean positive reaction strengths | ||||||
|---|---|---|---|---|---|---|
| -- | -- | -- | -- | -- | -- | -------------------------------------------------------------------------------------- |
10
| Table 4. Patient gender statistics and mean positive reaction strengths | |
|---|---|
| ------------------------------------------------------------------------- | -- |
| Male | Female | Not defined | |
|---|---|---|---|
| Clinical site 1 | |||
| Total tested | 70 | 124 | 0 |
| Mean positive reaction strength | 5.6 | 6.7 | N/A |
| Positive reaction range | 1-10 | 1-10 | N/A |
| Clinical site 2 | |||
| Total tested | 28 | 27 | 0 |
| Mean positive reaction strength | 8.2 | 7.9 | N/A |
| Positive reaction range | 1-10 | 1-10 | N/A |
| Clinical site 3 | |||
| Total tested | N/A | N/A | 400 |
| Mean positive reaction strength | N/A | N/A | 7 |
| Positive reaction range | N/A | N/A | 1-10 |
| Clinical site Totals | |||
| Total tested | 98 | 151 | 400 |
| Mean positive reaction strength | 6.6 | 7.1 | 7 |
| Positive reaction range | 1-10 | 1-10 | 1-10 |
Sample comparison
Semi-solid or liquid (diarrheal) stools are more frequently encountered in patients with C. difficileassociated disease (CDAD). However, it may be necessary to test solid stool for the presence of toxin. Instructions for collecting, preparing and storing the three stool types are given in the instructions for inditions for our of consoling for specimens used in the clinical trial were semi-solid. Appropriate results were obtained with all three types.
Table 5. Stool sample type and mean positive reaction strengths
| Stool Type | ||||
|---|---|---|---|---|
| Solid | Semi-solid | Liquid | Not defined | |
| Total tested -- Clinical Site 1 | ||||
| Total tested | 23 | 108 | 63 | 0 |
| Mean positive reaction strength | 7.0 | 5.6 | 6.7 | N/A |
| Positive reaction range | 1-10 | 1-10 | 1-10 | N/A |
| Total tested -- Clinical Site 2 | ||||
| Total tested | 20 | 19 | 16 | 0 |
| Mean positive reaction strength | 7.2 | 7.8 | 9.7 | N/A |
| Positive reaction range | 1-10 | 1-10 | 1-10 | N/A |
| Total tested -- Clinical Site 3 | ||||
| Total tested | 12 | 124 | 206 | 0 |
| Mean positive reaction strength | 3 | 7.5 | 7.1 | N/A |
| Positive reaction range | 1-3 | 1-9 | 1-10 | N/A |
| Total tested -- All Sites | ||||
| Total tested | 55 | 251 | 285 | 0 |
| Mean positive reaction strength | 6.9 | 6.9 | 7.1 | N/A |
| Positive reaction range | 1-10 | 1-10 | 1-10 | N/A |
11
| Table 6. Stool sample storage parameters and mean positive reaction strengths | |
|---|---|
| -------------------------------------------------------------------------------- | -- |
| Stool Type | |||
|---|---|---|---|
| Fresh | Frozen | Not recorded | |
| Total tested -- Clinical Site 1 | 142 | 52 | 0 |
| Total tested | 142 | 52 | 0 |
| Mean positive reaction strength | 6.6 | 5.7 | N/A |
| Positive reaction range | 1-10 | 1-10 | N/A |
| Total tested -- Clinical Site 2 | 0 | 55 | 0 |
| Total tested | 0 | 55 | 0 |
| Mean positive reaction strength | N/A | 8.0 | N/A |
| Positive reaction range | N/A | 1-10 | N/A |
| Total tested -- Clinical Site 3 | 0 | 342 | 0 |
| Total tested | 0 | 342 | 0 |
| Mean positive reaction strength | N/A | 7.1 | N/A |
| Positive reaction range | N/A | N/A | N/A |
| Total tested -- All Sites | 142 | 449 | 0 |
| Total tested | 142 | 449 | 0 |
| Mean positive reaction strength | 6.6 | 7.2 | N/A |
| Positive reaction range | 1-10 | 1-10 | N/A |
Clinical trial data summarized
The data collected during clinical trials is shown in the spreadsheets provided at the end of these sections. The results can be summarized as follows:
Table 7. Results of clinical evaluations
| Clinical site 1 | ICTAB | | | Reference
BD ColorPAC | | | Reference
Wampole | | |
|--------------------------------|------------------------------|-----|-------|--------------------------|-----|-------|----------------------|-----|-------|
| | Pos | Neg | Total | Pos | Neg | Total | Pos | Neg | Total |
| Cytotoxin Pos (Std) | 41 | 2 | 43 | 35 | 8 | 43 | 38 | 5 | 43 |
| Cytotoxin Neg (Std) | 1 | 150 | 151 | 2 | 149 | 151 | 6 | 145 | 151 |
| Total | 42 | 152 | 194 | 37 | 157 | 194 | 44 | 150 | 194 |
| | 95% CI | | | | | | | | |
| Clinical sensitivity | 41/43 (95.3%)
89.1-100% | | | 35/43 (81.4%) | | | 38/43 (88.4%) | | |
| Clinical specificity | 150/151 (99.3%)
97.4-100% | | | 149/151 (98.7%) | | | 145/151 (96.0%) | | |
| Predictive value positive test | 41/42 (97.6%)
94.1-100% | | | 35/37 (94.6%) | | | 38/44 (86.4%) | | |
| Predictive value negative test | 150/152 (98.5%)
97.4-100% | | | 149/157 (94.9%) | | | 145/150 (96.7%) | | |
| Clinical site 2 | ICTAB | Reference Oxoid | Reference PTAB | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Pos | Neg | Total | Pos | Neg | Total | Pos | Neg | Total | |
| Cytotoxin Pos (Std) | 25 | 3 | 28 | 25 | 3 | 28 | 26 | 2 | 28 |
| Cytotoxin Neg (Std) | 1 | 26 | 27 | 1 | 26 | 27 | 3 | 24 | 27 |
| Total | 26 | 29 | 55 | 26 | 29 | 55 | 29 | 26 | 55 |
| 95% CI | |||||||||
| Clinical sensitivity | 25/28 (89.3%) | 77.2-100% | 25/28 (89.3%) | 26/28 (92.9%) | |||||
| Clinical specificity | 26/27 (96.3%) | 88.2-100% | 26/27 (96.3%) | 24/27 (88.9%) | |||||
| Predictive value positive test | 25/26 (96.2%) | 88.2-100% | 25/26 (96.2%) | 26/29 (89.7%) | |||||
| Predictive value negative test | 26/29 (89.7%) | 78.2-100% | 26/29 (89.7%) | 24/26 (92.3%) |
12
| Clinical site 3 | ICTAB | | | Reference
BD | | |
|--------------------------------|-----------------|-----|------------|-----------------|-----|-------|
| | Pos | Neg | Total | Pos | Neg | Total |
| Cytotoxin Pos (Std) | 34 | 0 | 34 | 25 | 9 | 34 |
| Cytotoxin Neg (Std) | 5 | 303 | 308 | 11 | 297 | 308 |
| Total | 39 | 303 | 342 | 36 | 306 | 342 |
| | | | 95% CI | | | |
| Clinical sensitivity | 34/34 (100%) | | NA | 25/34 (71.4%) | | |
| Clinical specificity | 303/308 (98.3%) | | 96.4-99.6% | 297/308 (96.4%) | | |
| Predictive value positive test | 34/39 (87.2%) | | 75.2-98.8% | 25/36 (69.4%) | | |
| Predictive value negative test | 303/303 (100%) | | NA | 297/306 (97.1%) | | |
| Total Sites Combined Data - Comparison to Standard | ICTAB | |||
|---|---|---|---|---|
| Pos | Neg | Total | ||
| Cytotoxin Pos (Std) | 100 | 5 | 105 | |
| Cytotoxin Neg (Std) | 7 | 479 | 486 | |
| Total | 107 | 484 | 591 | |
| 95% CI | ||||
| Clinical sensitivity | 100/105 (95.2%) | 90.9-99.1% | ||
| Clinical specificity | 479/486 (98.5%) | 98.0-100% | ||
| Predictive value positive test | 100/107 (93.5%) | 88.1-97.9% | ||
| Predictive value negative test | 479/484 (99.0%) | 98.2-99.8% |
Combined data differentiated based on sample type
| Prospective Samples | ||||
|---|---|---|---|---|
| Pos | Neg | Total | ||
| Cytotoxin Pos (Std) | 67 | 5 | 72 | |
| Cytotoxin Neg (Std) | 2 | 176 | 178 | |
| Total | 69 | 181 | 250 | |
| 95% CI | ||||
| Clinical sensitivity | 67/72 (93.1%) | 87.1-98.9% | ||
| Clinical specificity | 176/178 (98.9%) | 97.6-100% | ||
| Predictive value positive test | 67/69 (97.1%) | 92.9-100% | ||
| Predictive value negative test | 176/181 (97.2%) | 94.5-99.5% | ||
| Retrospective Samples | ||||
| Pos | Neg | Total | ||
| Cytotoxin Pos (Std) | 33 | 0 | 33 | |
| Cytotoxin Neg (Std) | 5 | 303 | 308 | |
| Total | 38 | 303 | 341 | |
| 95% CI | ||||
| Clinical sensitivity | 33/33 (100%) | N/A | ||
| Clinical specificity | 303/308 (98.4%) | 96.4-99.6% | ||
| Predictive value positive test | 33/38 (86.8%) | 76.2-97.8% | ||
| Predictive value negative test | 303/303 (100%) | N/A |
13
Characterization of samples producing discordant results
Samples producing discordant test results between ImmunoCard Toxins A& B and cytotoxin are listed Samples producing ulscordant test results betwoor immaries and first and the positive below. Three of the hee samples oldobined as infracts that were not neutralized in the cytotoxin assay until the specimens were diluted 1 in 10.
| Sample
Number | ICTAB
Result | Cytotoxin
Result | Counted as
ICTAB | Comment |
|------------------|-----------------|---------------------|---------------------|-----------------------------------------------------------------------------------------------------|
| 1-32 | Neg | Pos | FN | No repeat testing performed |
| 1-119 | Neg | Pos | FN | No repeat testing performed |
| 1-270 | Pos | Neg | FP | No repeat testing performed |
| 2-1 | Neg | Pos | FN | No repeat testing performed |
| 2-2 | Neg | Pos | FN | No repeat testing performed |
| 2-25 | Neg | Pos | FN | No repeat testing performed |
| 2-41 | Pos | Neg | FP | No repeat testing performed |
| 3-103 | Pos | Neg | FP | Strong positive sample. Cytotoxin assay converted to positive
on repeat testing at 1:10 dilution |
| 3-118 | Pos | Neg | FP | Strong positive sample. Cytotoxin assay converted to positive
on repeat testing at 1:10 dilution |
| 3-200 | Pos | Neg | FP | Cytotoxin assay remained negative on repeated testing |
| 3-233 | Pos | Neg | FP | No repeat testing performed |
| 3-241 | Pos | Neg | FP | Strong positive sample. Cytotoxin assay converted to positive
on repeat testing at 1:10 dilution |
Table 8. Samples producing discrepant results.
Legend: ICTAB ≤ ImmunoCard Toxins A & B, FN = false negative, FP = false positive
Reproducibility
Reproducibility panels, consisting of eight coded specimens were sent to the three clinical sites. Five of these r reproducilly partified by the predicate device Premier Toxins A & B as positive and one was at the limit of stamples were stablined by the produce a positive or negative or negative result. Even though the trial sites were instructed to grade reactions, there were no criteria regarding the strength of a positive reaction that was expected. As the data shows, there was 100% reproducibility/precision within a test site and between the test sites.
| Clinical Site 1 | Clinical Site 2 | Clinical Site 3 | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Sample ID | Day 1 | Day 2 | Day 3 | Day 1 | Day 2 | Day 3 | Day 1 | Day 2 | Day 3 |
| 1 LP | 2 | 1 | 2 | 5 | 4 | 4 | 2 | 2 | 3 |
| 2 LP | 2 | 2 | 3 | 7 | 5 | 6 | 6 | 6 | 5 |
| 3 MP | 5 | 6 | 6 | 4 | 6 | 6 | 5 | 5 | 6 |
| 4 MP | 7 | 7 | 8 | 9 | 9 | 9 | 6 | 7 | 8 |
| 5 HP | 10 | 9 | 10 | 8 | 9 | 10 | 10 | 10 | 8 |
| 6 N | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 7 N | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 8 Limit of detect | 3 | 3 | 3 | 4 | 5 | 6 | 3 | 4 | 4 |
| Total positive score | 29 | 28 | 32 | 37 | 38 | 41 | 32 | 34 | 34 |
| Average positive score | 4.8 | 4.6 | 5.3 | 6.2 | 6.3 | 6.8 | 5.3 | 5.7 | 5.7 |
| Percent correlation | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 |
Table 9. Results with reproducibility test panels.
Legend: LP = low positive, MP = moderate positive, HP = high positive, N = negative,
14
Analytical sensitivity
The analytical sensitivity of the assay was determined in tests using negative stool samples that had The arraying of the about the about has spure toxin A or pure toxin B. The assay limit of detection is been i spitca will varying our centralit noduced a positive reaction of grade 1 or more across three lots of ImmunoCard. The results of these studies showed the assay limit of detection is 3 ng/mL for both toxin A and toxin B.
| ICTAB Lot | ICTAB Lot | ICTAB Lot | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| 712050.002 | 712050.003 | 712050.004 | ||||||||
| Toxin A | Control | Test | Interp | Control | Test | Interp | Control | Test | Interp | |
| ng/mL | ||||||||||
| 5.00 | 4 | 1/2 | Pos | 7 | 1/2 | Pos | 9 | 2 | Pos | |
| 4.00 | 4 | 1/2 | Pos | 7 | 1 | Pos | 9 | 2 | Pos | |
| 3.00 | 4 | 1 | Pos | 7 | 1 | Pos | 9 | 1 | Pos | |
| 2.00 | 4 | 1 | Pos | 7 | 1 | Pos | 9 | 1 | Pos | |
| 1.00 | 4 | 0 | Neg | 7 | 1 | Pos | 9 | 0/1 | Pos | |
| 0.50 | 4 | 0 | Neg | 7 | 0 | Neg | 9 | 0/1 | Pos | |
| 0.25 | 3 | 0 | Neg | 7 | 0 | Neg | 9 | 0 | Neg | |
| 0.00 | 2 | 0 | Neg | 7 | 0 | Neg | 9 | 0 | Neg | |
| PC | Pass | |||||||||
| NC | Pass | |||||||||
| Endpoint | 2.00 | 1.00 | 2.00 | |||||||
| Toxin B | ||||||||||
| ng/mL | ||||||||||
| 3.00 | 2 | 1 | Pos | 7 | 2 | Pos | 9 | 2 | Pos | |
| 2.00 | 2 | 1 | Pos | 7 | 2 | Pos | 9 | 2 | Pos | |
| 1.00 | 3 | 0/1 | Pos | 7 | 1/2 | Pos | 9 | 1/2 | Pos | |
| 0.50 | 3 | 0/1 | Pos | 7 | 0/1 | Pos | 9 | 1 | Pos | |
| 0.25 | 3 | 0 | Neg | 7 | 0 | Neg | 9 | 0/1 | Pos | |
| 0.125 | 3 | 0 | Neg | 7 | 0 | Neg | 9 | 0 | Neg | |
| 0.0625 | 3 | 0 | Neg | 7 | 0 | Neg | 9 | 0 | Neg | |
| 0.00 | 3 | 0 | Neg | 7 | 0 | Neg | 9 | 0 | Neg | |
| PC | Pass | |||||||||
| NC | Pass | |||||||||
| Endpoint | 2.00 | 1.00 | 0.50 |
Table 10. Limit of detection testing.
High dose hook effect
There was no high dose hook effect observed in verification or clinical testing performed with this assay.
Clinical trial data shows that ImmunoCard Toxins A & B is substantially equivalent to the standard method (cytotoxicity) and to predicate devices currently approved to market in the United States.
CONCLUSIONS
ImmunoCard Toxins A & B meets all performance claims when used to test hurnan stool specimens from the general population.
15
Image /page/15/Picture/1 description: The image shows the logo for the Department of Health & Human Services (HHS). The logo consists of a stylized human figure with three flowing lines representing health, services, and people. The figure is positioned to the right of the text "DEPARTMENT OF HEALTH & HUMAN SERVICES (USA)", which is arranged in a circular fashion around the figure.
Food and Drug Administration 2098 Gaither Road Rockville MD 20850
AUG 2 0 2004
Ms. Susan Rolih Vice President, Regulatory Affairs/Quality Assurance Meridian Bioscience, Inc. 3471 River Hills Drive Cincinnati, OH 45244
K041003 Re:
Trade/Device Name: Immuno Card® Toxins A & B Regulation Number: 21 CFR 866.2660 Regulation Name: Microorganism differentiation and identification device Regulatory Class: Class I Product Code: LLH Dated: July 21, 2004 Received: July 22, 2004
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).
16
Page 2
This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific 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 (301) 594-3084. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.
Sincerely yours,
Saaxtys
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
17
Meridian Bioscience, Inc. Cincinnati, OH
INDICATIONS FOR USE (Amended 8/14/04)
510(K) Number (if known):
Device Name: ImmunoCard Toxins A& B
Indications For Use: ImmunoCard Toxins A & B is an in vitro diagnostic qualitative enzyme immunoassay to detect the presence of Clostridium difficile toxins A and B in human stool. The assay is used as an aid in the diagnosis of C. difficile associated disease.
Prescription Use _____________________________________________________________________________________________________________________________________________________________ (Part 21 CFR 801 Subpart D)
. .
AND/OR
Over-The-counter Use (21 CFR 807 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
Ferdinand Carlo
Division Sign-Off
8-1
Office of In Vitro Diagnostic Device Evaluation and Safety
510(k) KC4 1003