(91 days)
Not Found
No
The description focuses on standard molecular diagnostic techniques (PCR and capillary electrophoresis) and does not mention any AI or ML components for data analysis or interpretation.
No.
This device is an in vitro diagnostic (IVD) intended to aid in the diagnosis of C. difficile infection by detecting the tcdB gene in stool specimens. It is not used to treat or prevent a disease or condition.
Yes
The device qualitatively detects the Clostridium difficile toxin B gene to aid in the diagnosis of C. difficile infection (CDI) in human stool specimens.
No
The device description explicitly states that the ICEPlex C. difficile Assay kit is intended to be used only on the ICEPlex System, which is described as combining a PCR thermal cycler (amplification module) and a capillary electrophoresis system with fluorescent detection (analysis module). These are hardware components. The assay kit itself also contains physical reagents (PCR enzyme, primer mix, buffers, controls). Therefore, the device is not software-only.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it's for the "qualitative detection of the Clostridium difficile toxin B gene (tcdB gene) in nucleic acids purified from unpreserved liquid or soft human stool specimens from patients suspected of having C. difficile infection (CDI)." This involves testing a sample taken from the human body to provide information about a disease state.
- Device Description: The device description details a kit used to perform a laboratory test (PCR and capillary electrophoresis) on human specimens.
- Performance Studies: The document describes clinical and analytical performance studies conducted to evaluate the device's ability to accurately detect the target in human samples.
- Comparison to Predicate Device: The mention of a predicate device (K130470 BD Max C. diff Assay) which is also an IVD, further supports this classification.
All these elements align with the definition of an In Vitro Diagnostic device, which is a medical device intended for use in vitro for the examination of specimens 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 compatibility of a transplant organ with a potential recipient, or to monitor therapeutic measures.
N/A
Intended Use / Indications for Use
The PrimeraDx ICEPlex® C. difficile Assay is for the qualitative detection of the Clostridium difficile toxin B gene (tcdB gene) in nucleic acids purified from unpreserved liquid or soft human stool specimens from patients suspected of having C. difficile infection (CDI).
The ICEPlex C. difficile Assay is intended to be used only on the ICEPlex® System, which integrates PCR-based amplification with capillary electrophoresis (CE) for the detection of amplification products. The assay is intended to aid in the diagnosis of CDI. Results should be considered in conjunction with patient clinical history.
Product codes (comma separated list FDA assigned to the subject device)
OZN, NSU
Device Description
PrimeraDx's ICEPlex C. difficile Assay kit is intended to be used only on the ICEPlex System. The ICEPlex C. difficile Assay kit incorporates several universal features and approaches developed at PrimeraDx for the ICEPlex System. The ICEPlex C. difficile Assay kit includes a PCR enzyme and the appropriate PCR buffer system developed, optimized, verified and validated for ideal performance in multiplex PCR with subsequent capillary electrophoresis. The ICEPlex C. difficile assay kit includes a primer mix for detection of the Clostridium difficile toxin B gene (tcdB gene) in human stool specimen from patients suspected of having C. difficile infection (CDI). The ICEPlex C. difficile assay kit is comprised of a PCR enzyme, primer mix, PCR buffer, calibrators mix, injection buffer, internal control, and a positive control.
The ICEPlex System combines two functional modules: an amplification module - PCR (Polymerase Chain Reaction) thermal cycler - and an analysis module -CE (Capillary Electrophoresis) system with fluorescent detection. Individual fluorescent PCR products from multiplexed PCR reactions are analyzed by CE through direct electrokinetic injection into the separating capillaries. The labeled amplicons are separated by size and the dyes are excited by two lasers within the system.
ICEPlex System Features
. Bench top instrument with PCR thermal cycler and integrated Capillary Electrophoresis (CE) system
Two solid state lasers enabling fluorescent detection of dye-labeled PCR . amplification products
On-board reagents with liquid level sensing .
. Touch screen software for easy, step-by-step processing of samples
. Ability to run multiple assays in the same plate
. Automated data analysis and result reporting
Simultaneous detection and quantification of multiple DNA targets in the . same reaction well.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
For prescription use only
Description of the training set, sample size, data source, and annotation protocol
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Description of the test set, sample size, data source, and annotation protocol
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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
a. ANALYTICAL PERFORMANCE
Precision
The ICEPlex C. difficile Assay Precision Study was performed on a panel of samples prepared by spiking an appropriate amount of C. difficile culture isolate from ATCC strain BAA-1805 into a pooled clinical negative stool matrix. The panel members included: Low positive, Moderately positive, Negative sample, (C20-80) sample. Runs for this study also include the following controls: Negative Control, Positive Control. The precision study consisted of two operators; three lots of ICEPlex C. difficile Assay kits; and three ICEPlex instruments. The precision study was run over 12 non-consecutive days with two runs per day and two replicates of each sample per run. One negative sample (out of a total of 144 observations) and one low positive sample (out of a total of 144 observations) produced invalid results during the study.
Reproducibility
Reproducibility of the ICEPlex C. difficile Assay was evaluated at 3 independent laboratory sites. The reproducibility study panel included 4 simulated samples - moderate positive (expected positive 100% of the time), low positive (near assay limit of detection, expected positive >95% of the time), negative (expected negative 100% of the time) and C20-80, ( expected positive 20-80% of the time). The panel also included positive and negative controls. Panel samples were tested at each independent laboratory site for 5 days with 2 runs per day and 3 replicates of each panel member per run. One low positive sample (out of a total of 90 observations) and one moderate positive sample (out of a total of 90 observations) produced invalid results during the study.
Analytical Sensitivity (Limit of Detection)
Analytical sensitivity of the ICEPlex C. difficile Assay was determined using a 2-fold serial dilution of two C. difficile strains that were spiked into qualified negative stool and processed according to ICEPlex C. difficile Assay Instructions for Use. 20 replicates at each concentration level were tested on 3 different ICEPlex instruments. Analytical sensitivity of the assay was defined as the lowest concentration at which at least 95% of all replicates were reported positive. LOD of the ICEPlex C. difficile assay determined for: C. difficile strain 43255 (630) (Toxinotype 0): 8CFU/rxn , C. difficile strain BAA-1805 (Toxinotype III): 2CFU/rxn.
Analytical Reactivity (Inclusivity)
To assess the analytical inclusivity of the ICEPlex C. difficile Assay, a set of 20 additional toxigenic strains of C. difficile were spiked into negative matrix (pool of negative clinical samples) at a level approximately three times above the assay LOD. Spiked samples were processed in accordance with the ICEPlex C. difficile Assay Instructions for Use. All strains tested positive.
Analytical Specificity - Cross Reactivity
A microbial cross reactivity study was performed with the ICEPlex C. difficile Assay using a panel of samples, consisting of pooled negative clinical matrix, spiked with 5 non-toxigenic C. difficile strains, 14 other Clostridium strains or 54 other pathogens or representatives of healthy intestinal flora. Bacteria and parasites were tested at concentrations 1x10^7 organisms/ml, viruses at concentrations 1x10^7 - 1x10^8 TCID50/ml. In this study, Clostridium sordellii was identified as cross-reacting, due to high toxin sequence similarity. Clostridium sordellii is not typically found in GI tract. Reactivity with this organism has little to no clinical significance. One of three replicates for Clostridium difficile strain 43601 was reported positive at 14 cps/rxn just above the assay cutoff set at 12 cps/rxn. PrimeraDx was not able to verify interference to this strain in follow-up analysis. None of other species resulted in positive result with ICEPlex C. difficile Assay.
Interfering Substances
A chemical interference study was performed using a panel of samples, consisting of pooled negative clinical matrix and contrived samples produced by supplementing pooled negative clinical matrix with culture stock of two C. difficile strains, ATCC BAA-1805 and 43255, added to produce samples resulting in 9 and 21 cfu/rxn, respectively (approximately three times the assay LOD). None of the substances interfered with detection of C. difficile from both tested strains or caused a false positive result in the negative samples.
Contamination and Well-to-well Cross Run-to-run Carryover Contamination
The study identified no contamination in the negative samples run in the course of the study, reporting False Positive rate at 0% (0 False Positives out of 141 valid negatives). These data allowed PrimeraDx to conclude that there was no well-to-well Cross Contamination or run-to-run Carryover Contamination observed in the course of this study.
b. CLINICAL PERFORMANCE STUDIES
A clinical study was conducted at three independent sites to compare performance of the ICEPlex C. difficile Assay Kit on the ICEPlex System to toxigenic C. difficile direct culture.
This study protocol directed the laboratory testing of specimens from patients suspected of gastrointestinal tract infection with a toxigenic strain of Clostridium difficile bacteria. Each of the sites performed testing in the ICEPlex C. difficile Assay on specimens collected at the site. These specimens collected from all three sites were tested by toxigenic C. difficile direct culture. A total of 1103 (806 fresh, 297 frozen) samples were collected and enrolled in the investigational study at 3 clinical sites. 97 frozen samples were excluded from the analysis due to improper storage conditions and temperature excursions. An additional 37 samples were excluded due to study protocol deviations.
969 specimens were compliant and met all protocol requirements. 952 (98.2%) out of 969 gave reportable results and were included in the statistical analysis. 17 samples remained invalid upon retest (1.8% of all analyzed samples). 3 out of the 17 unresolved invalids were reported positive by direct culture.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Positive Percent Agreement (95% CI): 90.0% ( 84.5 - 94.1)
Negative Percent Agreement (95% CI): 97.4% ( 96.1 - 98.4)
PPA (95% CI) (Site A): 49/55 = 89.1% (77.8 - 95.9)
NPA (95% CI) (Site A): 281/288 = 97.6% (95.1 - 99.0)
PPA (95% CI) (Site B): 56/64 = 87.5% (76.8 - 94.4)
NPA (95% CI) (Site B): 256/262 = 97.7% (95.1 - 99.2)
PPA (95% CI) (Site C): 48/51 = 94.1% (83.8 - 98.8)
NPA (95% CI) (Site C): 225/232 = 97.0% (93.9 - 98.8)
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.
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
§ 866.3130 Clostridium difficile toxin gene amplification assay.
(a)
Identification. AClostridium difficile toxin gene amplification assay is a device that consists of reagents for the amplification and detection of target sequences inClostridium difficile toxin genes in fecal specimens from patients suspected of havingClostridium difficile infection (CDI). The detection of clostridial toxin genes, in conjunction with other laboratory tests, aids in the clinical laboratory diagnosis of CDI caused byClostridium difficile. (b)
Classification. Class II (special controls). The special controls are set forth in FDA's guideline document entitled: “Class II Special Controls Guideline: Toxin Gene Amplification Assays for the Detection ofClostridium difficile; Guideline for Industry and Food and Drug Administration Staff.” See § 866.1(e) for information on obtaining this document.
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510(k) Summary
ICEPlex C. difficile Kit on the ICEPlex System
A. 510(K) NUMBER: K132726
Date Prepared: November 29, 2013
- B. SUBMITTED BY (APPLICANT):
PrimeraDx
171 Forbes Blvd., Suite 1000 Mansfield, MA 02048
NOV 29 2013
Contact Person
Fayyaz Memon Vice President, Regulatory Affairs and Quality Assurance Tel: 301-221-0245 Fax: 508-339-0452 Email: fmemon@primeradx.com
C. PURPOSE OF SUBMISSION
Substantial equivalence determination of ICEPlex C. difficile Assay Kit on the ICEPlex® System for qualitative detection of the C. difficile toxin B gene (tcdB gene) in a human stool specimen in patients suspected of having C. difficile infection (CDI).
D. Measurand
Clostridium difficile toxin B gene (tcdB)
E. TYPE OF TEST:
Qualitative Nucleic Acid Amplification Test for C. difficile toxin B gene from liquid or soft tool specimen.
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F. PROPRIETARY AND ESTABLISHED NAMES (TRADE NAMES)
Assay: ICEPlex C. difficile Assay
System: ICEPlex® System
G. REGULATORY INFORMATION
a. Regulatory Section:
21 CFR 866.3130 - C. difficile Nucleic Acid Amplification Test Assay
b. Classification:
Class II
c. Product Code
OZN, NSU
d. Panel
Microbiology (83)
H. INTENDED USE
a. Intended Use
The PrimeraDx ICEPlex® C. difficile Assay is for the qualitative detection of the Clostridium difficile toxin B gene (tcdB gene) in nucleic acids purified from unpreserved liquid or soft human stool specimens from patients suspected of having C. difficile infection (CDI).
The ICEPlex C. difficile Assay is intended to be used only on the ICEPlex® System, which integrates PCR-based amplification with capillary electrophoresis (CE) for the detection of amplification products. The assay is intended to aid in the diagnosis of CDI. Results should be considered in conjunction with patient clinical history.
b. Indication(s) for use
Same as Intended Use
2
c. Special Conditions for use statement(s)
For prescription use only
d. Special Instrument Requirement
ICEPlex® System
I. DEVICE DESCRIPTION
PrimeraDx's ICEPlex C. difficile Assay kit is intended to be used only on the ICEPlex System. The ICEPlex C. difficile Assay kit incorporates several universal features and approaches developed at PrimeraDx for the ICEPlex System. The ICEPlex C. difficile Assay kit includes a PCR enzyme and the appropriate PCR buffer system developed, optimized, verified and validated for ideal performance in multiplex PCR with subsequent capillary electrophoresis. The ICEPlex C. difficile assay kit includes a primer mix for detection of the Clostridium difficile toxin B gene (tcdB gene) in human stool specimen from patients suspected of having C. difficile infection (CDI). The ICEPlex C. difficile assay kit is comprised of a PCR enzyme, primer mix, PCR buffer, calibrators mix, injection buffer, internal control, and a positive control.
| # of vials and volume | |
|---|---|
| 2 vials, 1.5ml/vial | 2x PCR Buffer |
| P/N: 560-1001 | |
| 1 vial, 45µl/vial | PCR Enzyme |
| P/N: 560-1002 | |
| 1 vial, 220µl/vial | 25x Primer Mix |
| P/N: 560-1003 | |
| 1 vial, 220 µl/vial | 25x Calibrators Mix |
| P/N: 560-1004 | |
| 1 vial, 100µl/vial | Positive Control |
| P/N: 560-1005 | |
| 3 vials, 1.5 ml/vial | 10x Injection Buffer |
| P/N: 560-1006 | |
| 1 vial, 1.0 ml/vial | Internal Control |
| P/N: 560-1007 |
ICEPlex C. difficile Assay kit components
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The ICEPlex System combines two functional modules: an amplification module - PCR (Polymerase Chain Reaction) thermal cycler - and an analysis module -CE (Capillary Electrophoresis) system with fluorescent detection. Individual fluorescent PCR products from multiplexed PCR reactions are analyzed by CE through direct electrokinetic injection into the separating capillaries. The labeled amplicons are separated by size and the dyes are excited by two lasers within the system.
ICEPlex System Features
- . Bench top instrument with PCR thermal cycler and integrated Capillary Electrophoresis (CE) system
- Two solid state lasers enabling fluorescent detection of dye-labeled PCR . amplification products
- On-board reagents with liquid level sensing .
- . Touch screen software for easy, step-by-step processing of samples
- . Ability to run multiple assays in the same plate
- . Automated data analysis and result reporting
- Simultaneous detection and quantification of multiple DNA targets in the . same reaction well.
J. SUBSTANTIAL EQUIVALENCE COMPARISON
a. PREDICATE DEVICE NAME BD Max C. diff Assay
b. PREDICATE DEVICE 510(K) NUMBER K130470
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| PrimeraDx | Predicate: | |
|---|---|---|
| Similarities | ||
| Intended | ||
| Use | The PrimeraDx ICEPlex® C. | |
| difficile Assay is for the | ||
| qualitative detection of the | ||
| Clostridium difficile toxin B | ||
| gene ( tcdB gene) in nucleic | ||
| acids purified from unpreserved | ||
| liquid or soft human stool | ||
| specimens from patients | ||
| suspected of having C. difficile | ||
| infection (CDI). The ICEPlex | ||
| C. difficile Assay is intended to | ||
| be used only on the ICEPlex® | ||
| System, which integrates PCR- | ||
| based amplification with | ||
| capillary electrophoresis (CE) | ||
| for the detection of | ||
| amplification products. The | ||
| assay is intended to aid in the | ||
| diagnosis of CDI. Results | ||
| should be considered in | ||
| conjunction with patient clinical | ||
| history. | The BD MAX C. diff Assay | |
| performed on the BD MAX | ||
| System is an automated in vitro | ||
| diagnostic test for the direct, | ||
| qualitative detection of the | ||
| Clostridium difficile toxin B | ||
| gene ( tcdB ) in human liquid or | ||
| soft stool specimens from | ||
| patients suspected of having C. | ||
| difficile infection (CDI). | ||
| The test, performed directly on | ||
| the specimen, utilizes real-time | ||
| polymerase chain reaction | ||
| (PCR) for the amplification of | ||
| C. difficile toxin B gene DNA | ||
| and fluorogenic target-specific | ||
| hybridization probes for the | ||
| detection of the amplified DNA. | ||
| The BD MAX C. diff Assay is | ||
| intended to aid in the diagnosis | ||
| of CDI. (Similar) | ||
| Measurand | Clostridium difficile | |
| toxin B gene ( tcdB ) | Same | |
| Specimen | ||
| Type | Unformed (liquid or | |
| soft stool) | Same | |
| Principle | DNA: real time PCR | Same |
| PrimeraDx (ICEPlex) | Predicate: | |
| Differences | ||
| Instrument | ||
| System | ICEPlex System | BD Max System |
| Detection/ | ||
| Probes | Assays use real time PCR | |
| amplification with Capillary | ||
| Electrophoresis and direct | ||
| laser-induced fluorescent | ||
| detection of target-specific | ||
| labeled primers in optical | ||
| channels. | Assay use PCR and | |
| amplified DNA targets are detected | ||
| using hydrolysis (TaqMan) probes. | ||
| Probes labeled flourophores are | ||
| used | ||
| to detect tcdB. Emitted fluorescent | ||
| is | ||
| measured in optical channels. | ||
| Sample | ||
| Extraction | bioMérieux NucliSENS | |
| easyMAG | Integrated | |
| Test | ||
| Cartridge | Disposable single-use | |
| PCR plate and micro titer | ||
| plate, and ICEPlex Cartridge | ||
| with 48 | ||
| Capillaries. | Disposable tray with reagents, | |
| microfluidics PCR chamber | ||
| Process | Extraction, PCR | |
| amplification, capillary | ||
| electrophoresis, size- based | ||
| target detection | Extraction, PCR | |
| amplification, in- sample | ||
| optical detection based on | ||
| hydrolysis probes. | ||
| Time to | ||
| result | Four hours for 48 | |
| samples | Under 3 hours for 24 samples | |
| Assay | ||
| Controls | Calibration Control, | |
| Internal control. PCR Positive | ||
| control provided. Negative | ||
| control and Positive/Extraction | ||
| Control provided by user | Sample processing control. User- | |
| provided Positive and negative | ||
| controls are recommended | ||
| Negative | Negative | |
| Results | Positive | |
| Invalid, with error code | Positive | |
| Unresolved - specimen | ||
| failure | ||
| Indeterminate - system | ||
| failure with error code | ||
| Incomplete Run - with | ||
| error code |
Comparison with Predicate Devices Showing Similarities
. .
5
Comparison with Predicate Devices Showing Difference
6
K. TEST PRINCIPLE
The ICEPlex C. difficile Assay on the ICEPlex System is a molecular diagnostic test for the qualitative detection of toxigenic C. difficile nucleic acids isolated and purified from liquid or soft stool specimens obtained from symptomatic patients. This test targets the C. difficile toxin B encoding gene (tcdB). The ICEPlex C. difficile Assay has been developed for use on the PrimeraDx ICEPlex System.
This instrument platform integrates PCR-based amplification with capillary electrophoresis (CE) based detection of amplification products. Oligonucleotide primers are designed to produce PCR products with unique CE mobility, enabling simultaneous measurement of multiple targets and controls in a single reaction. Each individual test includes three target sites of the tcdB gene (not reported separately), an Internal Control) and Calibrators. Calibrators of three sizes are used for aligning and assigning CE peaks, a procedure unique to the ICEPlex System. The ICEPlex C. difficile Assay is PCR based and is performed on nucleic acids purified from human stool sample. The ICEPlex C. difficile Assay is intended to be used only on the ICEPlex System.
To perform the test, liquid or soft stool is collected in a standard sterile container that can be sealed. Sample could be stored at 2-8 C for up to 48 hours prior to testing or frozen at -70°C or below if not processed within 48 hours. Sample extraction is performed using bioMérieux NucliSENS easyMAG™ System as per the ICEPlex C. difficile Assay instruction for use. PCR reagent master mix is prepared using a Primers mix, PCR enzyme, PCR buffer and calibrators. In a designated well of PCR plate, 40 uL of ICEPlex C. difficile Master Mix is aliquoted and 10 uL of extracted sample is added. Negative and positive controls are added in designated wells of the PCR plate. After the PCR plate is prepared, it is placed in the ICEPlex System to execute an instrument run for the detection of the Clostridium difficile toxin B gene (tcdB gene)per the user manual.
Assay Controls:
Controls provided with the ICEPlex C. difficile assay kit include:
Internal Control - Non-target nucleic acid that is co-extracted and coamplified with the tcdB target. It controls for nucleic acid extraction efficiency, for the integrity of the reagents and for the presence of PCR
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inhibitors in a given sample. The Internal Control needs to be spiked into each sample before extraction.
Calibration Control - A group of ICEPlex specific elements used to align electropherograms and assign identities of the target peaks. It also controls for the integrity of the kit reagents .
Positive Control - Non-infectious and non-contagious double stranded DNA containing fragment of the C. difficile tcdB gene.
Controls not provided with the ICEPlex C. difficile assay kit but required include:
Negative Control - Substitute S.T.A.R. buffer for the clinical specimen and process normally through the extraction system and on the ICEPlex Instrument.
Known Positive Sample - It is also required to include previously characterized positive sample or simulated sample with every easyMAG extraction run and include it in the subsequent ICEPlex Instrument run to verify successful lysis.
L. PERFORMANCE CHARACTERISTICS
a. ANALYTICAL PERFORMANCE
Precision
.
·
. . . . .
The ICEPlex C. difficile Assay Precision Study was performed on a panel of samples prepared by spiking an appropriate amount of C. difficile culture isolate from ATCC strain BAA-1805 into a pooled clinical negative stool matrix. The panel members included:
- . Low positive
- Moderately positive .
- Negative sample .
- (C20-80) sample. .
Runs for this study also include the following controls:
- Negative Control .
- Positive Control .
The precision study consisted of two operators; three lots of ICEPlex C. difficile Assay kits; and three ICEPlex instruments. The precision study was
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run over 12 non-consecutive days with two runs per day and two replicates of each sample per run. One negative sample (out of a total of 144 observations) and one low positive sample (out of a total of 144 observations) produced invalid results during the study.
| Concentration | Overall Agreement | Ct Value | |||
|---|---|---|---|---|---|
| Mean Ct | SD | %CV | |||
| Negative | 143/143 | 100% | N/A | N/A | N/A |
| Low Positive | 142/143 | 99% | 30.6 | 1.4 | 4.5 |
| Moderate Positive | 144/144 | 100% | 28.7 | 0.8 | 2.7 |
| Hi/Low (C20-80) | 105/144 | 73% | 34.6 | 2.1 | 6.2 |
Precision Study Overall Results
Reproducibility
Reproducibility of the ICEPlex C. difficile Assay was evaluated at 3 independent laboratory sites. The reproducibility study panel included 4 simulated samples - moderate positive (expected positive 100% of the time), low positive (near assay limit of detection, expected positive >95% of the time), negative (expected negative 100% of the time) and C20-80, ( expected positive 20-80% of the time). The panel also included positive and negative controls. Panel samples were tested at each independent laboratory site for 5 days with 2 runs per day and 3 replicates of each panel member per run. One low positive sample (out of a total of 90 observations) and one moderate positive sample (out of a total of 90 observations) produced invalid results during the study Study results are summarized in the tables below:
Reproducibility Study Result - Overall Agreement
| Observed | Total | % Agreement | 95% CI | |
|---|---|---|---|---|
| C20-80* | 62/90 | 90 | 69 | 58.26 to 78.23 |
| Moderate Positive | 89/89 | 89 | 100 | 95.98 to 100 |
| Low Positive | 89/89 | 89 | 100 | 95.98 to 100 |
| Negative | 90/90 | 90 | 100 | 95.98 to 100 |
| Negative control | 60/60 | 60 | 100 | 94.04 to 100 |
| Positive control | 60/60 | 60 | 100 | 94.04 to 100 |
NOTE: *For C20-80 samples % agreement is given as % positive results
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| Site A | Site B | Site C | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Observed | Total | % Agreement | Observed | Total | % Agreement | Observed | Total | % Agreement | |
| C20-80* | 23 | 30 | 77 | 23 | 30 | 77 | 16 | 30 | 53 |
| Moderate | |||||||||
| Positive | 30 | 30 | 100 | 30 | 30 | 100 | 29 | 29 | 100 |
| Low | |||||||||
| Positive | 30 | 30 | 100 | 29 | 29 | 100 | 30 | 30 | 100 |
| Negative | 30 | 30 | 100 | 30 | 30 | 100 | 30 | 30 | 100 |
| Negative | |||||||||
| Control | 20 | 20 | 100 | 20 | 20 | 100 | 20 | 20 | 100 |
| Positive | |||||||||
| Control | 20 | 20 | 100 | 20 | 20 | 100 | 20 | 20 | 100 |
Reproducibility Study, Site to Site % Agreement
Analytical Sensitivity (Limit of Detection)
Analytical sensitivity of the ICEPlex C. difficile Assay was determined using a 2-fold serial dilution of two C. difficile strains that were spiked into qualified negative stool and processed according to ICEPlex C. difficile Assay Instructions for Use. 20 replicates at each concentration level were tested on 3 different ICEPlex instruments. Analytical sensitivity of the assay was defined as the lowest concentration at which at least 95% of all replicates were reported positive.
LOD of the ICEPlex C. difficile assay determined for:
- . C. difficile strain 43255 (630) (Toxinotype 0): 8CFU/rxn
- . C. difficile strain BAA-1805 (Toxinotype III): 2CFU/rxn
Analytical Reactivity (Inclusivity)
To assess the analytical inclusivity of the ICEPlex C. difficile Assay, a set of 20 additional toxigenic strains of C. difficile were spiked into negative matrix (pool of negative clinical samples) at a level approximately three times above the assay LOD. Spiked samples were processed in accordance with the ICEPlex C. difficile Assay Instructions for Use. The results of the study are as follows:
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| Strain | Toxinotype | Result |
|---|---|---|
| BAA-1382 (630) | A+B+ | Positive |
| BAA-1871 (4111) | 0, A+B+binary-, NAP5 | Positive |
| 9689 (90556-M6S) | 0 | Positive |
| 700792 (14797-2) | A+B+ | Positive |
| BAA-1875 (5325) | V, A+B+, NAP7 | Positive |
| 51695 (BDMS 18 | ||
| AN) | A+B+ | Positive |
| 43598 (1470) | VIII, A-B+ | Positive |
| 43600 (2149) | A+B+ | Positive |
| 43599(2022) | A+B+ | Positive |
| 43597 | A+B+ | Positive |
| 43594 (W1194) | A+B+ | Positive |
| 43596 (545) | I, A+B+ | Positive |
| 17858 (1253) | A+B+ | Positive |
| 17857 (870) | A+B+ | Positive |
| BAA-1808 | A+B+ | Positive |
| BAA-1806 | A+B+ | Positive |
| BAA-1803 | III A+B+, NAP1 | Positive |
| BAA-1870 (4118) | III, binary+, NAP1 | Positive |
| BAA-1873 (5283) | 0, A+,B+,binary- | Positive |
: ・ ・ ・
۔ ﺑ
. --
Note: Strain BAA-1814 (Toxinotype XXII) was determined to be nonviable. PrimeraDx cannot claim inclusivity to this strain
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Analytical Snecificity
Cross Reactivity
A microbial cross reactivity study was performed with the ICEPlex C. difficile Assay using a panel of samples, consisting of pooled negative clinical matrix. spiked with 5 non-toxigenic C. difficile strains, 14 other Clostridium strains or 54 other pathogens or representatives of healthy intestinal flora. Bacteria and parasites were tested at concentrations 1x10' organisms/ml, viruses at concentrations1x10'-- 1x10° TCID50/ml. In this study, Clostridium sordellii was identified as cross-reacting, due to high toxin sequence similarity. Clostridium sordellii is not typically found in GI tract. Reactivity with this organism has little to no clinical significance. One of three replicates for Clostridium difficile strain 43601 was reported positive at 14 cps/rxn just above the assay cutoff set at 12 cps/rxn. PrimeraDx was not able to verify interference to this strain in follow-up analysis. None of other species resulted in positive result with ICEPlex C. difficile Assay.
Interfering Substances
A chemical interference study was performed using a panel of samples, consisting of pooled negative clinical matrix and contrived samples produced by supplementing pooled negative clinical matrix with culture stock of two C. difficile strains, ATCC BAA-1805 and 43255, added to produce samples resulting in 9 and 21 cfu/rxn, respectively (approximately three times the assay LOD). The table below shows potential interfering substances and concentrations used in this study.
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| Substance
| ్రామెన్ నేని వేసా ప్రసి | Active Ingredient | Concentration |
|---|---|---|
| Anti-Fungal /Anti-Itch Vaginal | Nystatin | 1% (w/v) |
| Creams/Ointments/Suppositories | Hydrocortisone | 1% (w/v) |
| Anti-Hemorrhoid Creams/Ointments | Phenylephrine | 1% (w/v) |
| Antacids | Calcium Carbonate/ | |
| Aluminum | ||
| Hydroxide/ | ||
| Magnesium | ||
| Hydroxide | 10% (w/v) | |
| Enemas | Mesalazine/Mineral | |
| Oil | 10% (w/v) | |
| Condoms with Spermicidal Lubricant | Nonoxynol-9 | 1% (w/v) |
| Anti-Diarrheal Medication | Loperamide | |
| Hydrochloride/ | ||
| Bismuth | ||
| Subsalicylate | 10% (w/v) | |
| Laxatives | Sennosides | 1% (w/v) |
| Antibiotic | Metronidazole | 12.5mg/ml |
| Antibiotic | Vancomycin | 12.5mg/ml |
| Non-Steroidal Anti-Inflammatory | ||
| Medications | Naproxen Sodium | 12.5mg/ml |
| Moist Towelettes | Benzalkonium | |
| Chloride, Ethanol | ().1% (v/v), | |
| 1%(v/v) | ||
| Fecal Fat | Lipids, etc. | 40% w/v |
| Whole Blood | Glucose, Hormones, | |
| Enzymes, Ions, Iron, | ||
| etc. | 40% v/v | |
| Mucus | Mucin protein | 3.5% (w/v) |
None of the substances interfered with detection of C. difficile from both tested strains or caused a false positive result in the negative samples.
Contamination and Well-to-well Cross Run-to-run Carryover Contamination
Samples for Well-to-well Cross Contamination and Run-to-run Carryover Contamination study were prepared by the extraction of a series of high positive samples (with analyte concentration exceeding the concentration found in 95% positive samples in the intended use population) alternating with negative samples. Location of positive and negative samples on the extraction instrument was altered
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run-to-run. On the ICEPlex system, high positive and negative samples were run in a checkerboard fashion. In the consecutive run, the platemap was inverted to allow wells and capillaries that were running negative samples to run positive samples. The study included 6 runs: each run had 24 high positive and 24 negative samples. The study identified no contamination in the negative samples run in the course of the study, reporting False Positive rate at 0% (0 False Positives out of 141 valid negatives). These data allowed PrimeraDx to conclude that there was no well-towell Cross Contamination or run-to-run Carryover Contamination observed in the course of this study.
b. CLINICAL PERFORMANCE STUDIES
A clinical study was conducted at three independent sites to compare performance of the ICEPlex C. difficile Assay Kit on the ICEPlex System to toxigenic C. difficile direct culture.
This study protocol directed the laboratory testing of specimens from patients suspected of gastrointestinal tract infection with a toxigenic strain of Clostridium difficile bacteria. Each of the sites performed testing in the ICEPlex C. difficile Assay on specimens collected at the site. These specimens collected from all three sites were tested by toxigenic C. difficile direct culture. A total of 1103 (806 fresh, 297 frozen) samples were collected and enrolled in the investigational study at 3 clinical sites. 97 frozen samples were excluded from the analysis due to improper storage conditions and temperature excursions. An additional 37 samples were excluded due to study protocol deviations.
969 specimens were compliant and met all protocol requirements. 952 (98.2%) out of 969 gave reportable results and were included in the statistical analysis. 17 samples remained invalid upon retest (1.8% of all analyzed samples). 3 out of the 17 unresolved invalids were reported positive by direct culture.
Performance characteristics determined in the course of this study were as follows. The Clopper-Pearson exact method was used to calculate confidence intervals.
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Overall Results
| Direct Culture | |||
|---|---|---|---|
| Positive | Negative | ||
| ICEPlex C. | |||
| difficile Assay | Positive | 153 | 20a |
| Negative | 17b | 762 | |
| Total: | 170 | 782 |
Positive Percent Agreement (95% CI): 90.0% ( 84.5 - 94.1) Negative Percent Agreement (95% CI): 97.4% ( 96.1 - 98.4)
Notes:
Discordant testing was performed for samples where ICEPlex C. difficile Assay and toxigenic C. difficile direct culture reported results in disagreement.
Discordant analysis included microbiological isolation of and PCR targeting of 3 appropriate regions of the toxin B gene (Different recognition sites than the ones used in the ICEPlex C. difficile assay) with bi-directional DNA sequencing.
46 of 20 reported positive by ICEPIex C. difficile Assay were reported positive by discordant analysis.
b14 of 17 reported negative by ICEPlex C. difficile Assay samples were reported positive by discordant analysis.
| | Site
A | Site
B | Site C |
|-----------------|----------------------------------|----------------------------------|----------------------------------|
| PPA
(95% CI) | 49/55 = 89.1%
(77.8 - 95.9) | 56/64 = 87.5%
(76.8 - 94.4) | 48/51 = 94.1%
(83.8 - 98.8) |
| NPA
(95% CI) | 281/288 = 97.6%
(95.1 - 99.0) | 256/262 = 97.7%
(95.1 - 99.2) | 225/232 = 97.0%
(93.9 - 98.8) |
Results by Site
M. CONCLUSION
PrimeraDx believes that based on the clinical and analytical performance comparison data, ICEPlex C. difficile Assay on the ICEPlex System is substantially equivalent to predicate devices.
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Image /page/15/Picture/0 description: The image shows a logo with a stylized bird-like figure composed of three curved lines, suggesting movement or flight. The logo is encircled by text, which appears to be part of the organization's name or a related phrase. The text is arranged in a circular fashion around the bird symbol, indicating a formal or official emblem.
DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring. MI) 20995-0002
PRIMERADX, INC. FAYYAZ MEMON VP. REGULATORY AFFAIRS AND QUALITY ASSURANCE 171 FORBES BLVD SUITE 1000 MANSFIELD MA 02048
November 29, 2013
Re: K132726
Trade/Device Name: ICEPlex C. difficile Assav Kit. ICEPlex System Regulation Number: 21 CFR § 866.3130 Regulation Name: Clostridium difficile toxin gene amplification assay Regulatory Class: II Product Code: OZN, NSU Dated: September 6, 2013 Received: September 9. 2013
Dear Mr. Memon:
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. Iisting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warrantics. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA). it may be rise is easilitional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations. Title 21. Parts 800 to 898. 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 I Teast be advised mar 1971 3 lessantes over device complies with other requirements of the Act many Federal statutes and regulations administered by other Federal agencies. You must or and 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); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803): good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CHR Part 820); and if applicable. the electronic product radiation control provisions (Sections 531-542 of the Act): 21 CFR 1000-1050.
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Page 2—Mr. Memon
If you desire specific advice for your device on our labeling regulations (2) CFR Parts 801 and 809). please contact the Division of Small Manufacturers. International and Consumer Assistance at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://yww.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803). please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRF' s Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on vour responsibilities under the Act from the Division of Small Manufacturers. International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm.
Sincerely yours.
Uwe Scherf -5 for
Sally A. Hojvat. M.Sc.. Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number: K132726
Device Name: ICEPlex®C. difficile Assay on the ICEPlex® System
Indications For Use:
The PrimeraDx ICEPlex® C. difficile Assay is for the qualitative detection of the Clostridium difficile toxin B gene (tcdB gene) in nucleic acids purified from unpreserved liquid or soft human stool specimens from patients suspected of having C. difficile infection (CDI).
The ICEPlex C. difficile Assay is intended to be used only on the ICEPlex® System, which integrates PCR-based amplification with capillary electrophoresis (CE) for the detection of amplification products. The assay is intended to aid in the diagnosis of CDI. Results should be considered in conjunction with patient clinical history.
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)
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Concurrence of CDRH; Office of In Vitro Diagnostics and Radiological Health (OIR)
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