(263 days)
No
The description details a system based on PCR and MALDI-TOF MS analysis with software that analyzes mass spectra using a "test-specific algorithm." There is no mention of AI, ML, or any learning-based approach.
No
The device is an in vitro diagnostic (IVD) device used for genetic testing to aid in the diagnosis of suspected thrombophilia, not for direct treatment or therapy.
Yes
The "Intended Use / Indications for Use" states that the device "is indicated for use as an aid in the diagnosis of patients with suspected thrombophilia."
No
The device is part of a system that includes significant hardware components (IMPACT Dx NANO, IMPACT Dx MA mass spectrometer) and reagents, in addition to the software. While the software is crucial for analysis, it is not a standalone software-only medical device.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is a "qualitative in vitro diagnostic device intended for use in the detection and genotyping..." This is the most direct indicator.
- Sample Type: The device uses "genomic DNA isolated from EDTA anti- coagulated human whole blood samples." This is a biological sample taken from the human body, which is characteristic of IVDs.
- Purpose: The test is indicated for use "as an aid in the diagnosis of patients with suspected thrombophilia." IVDs are used to provide information for the diagnosis, treatment, or prevention of disease.
- Device Description: The description details a process involving "genomic DNA purified from human whole blood" and analysis of "reaction products of the target nucleic acids." This aligns with the nature of in vitro diagnostic testing.
- Performance Studies: The document includes performance studies using "clinical samples" and comparing results to a "reference method," which are standard practices for evaluating the performance of IVDs.
- Predicate Devices: The mention of predicate devices with K numbers (e.g., K033607, K033612) indicates that this device is being compared to other legally marketed IVDs.
All of these points strongly support the classification of this device as an In Vitro Diagnostic.
N/A
Intended Use / Indications for Use
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is a qualitative in vitro diagnostic device intended for use in the detection and genotyping of a single point mutation (G1691A, referred to as the Factor V Leiden mutation or FVL) of the Factor V gene, located on Chromosome 1q23, and a single point mutation (G20210A) of the prothrombin gene (referred to as Factor II or FII), located on Chromosome 11p11-q12, from genomic DNA isolated from EDTA anti-coagulated human whole blood samples. The test is to be performed on the IMPACT Dx System and is indicated for use as an aid in the diagnosis of patients with suspected thrombophilia.
Product codes
PHJ
Device Description
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is a qualitative, multiplexed genetic testing device for parallel detection and genotyping of the point mutations G1691A of the Factor V gene and G20210A of the Factor II gene from genomic DNA isolated from EDTA anti- coagulated human whole blood samples. The test is to be performed on the IMPACT Dx System.
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is performed using the IMPACT Dx System, which includes the IMPACT Dx NANO and the IMPACT Dx MA, a matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometer. The test involves Factor V and Factor II region-specific polymerase chain reaction (PCR) amplification of genomic DNA purified from human whole blood in a multiplexed reaction, followed by allele-specific single base primer extension reactions. The reaction products are desalted, dispensed onto a SpectroCHIP® Array using the IMPACT Dx NANO, and the genotyping products are resolved on the basis of mass using the IMPACT Dx MA.
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test provides reagents for multiplex PCR, deoxynucleotide triphosphate dephosphorylation, and single base extension. The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is comprised of the following components:
- IMPACT Dx Factor V Leiden and Factor II Primer Set
- IMPACT Dx PCR Reagent Set
- IMPACT Dx Extend Reagent Set
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test utilizes a biochemistry process (Sequenom Biochemistry) that involves target-specific PCR amplification and single-base extension reactions with the subsequent analysis of the reaction products of the target nucleic acids by matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS),
IMPACT Dx System Overview
The IMPACT Dx System (System) is a platform for highly accurate and sensitive genomic analysis and is designed for use with FDA cleared or approved assays citing its use. The IMPACT Dx System is comprised of the following instruments, software and consumables;
- IMPACT Dx NANO (NANO)
- IMPACT Dx MA (MA)
- TYPER Dx Software (TYPER Dx)
- System Consumables
- SpectroCHIP® Arrays (Chip)
- Clean Resin
- 3-Point Calibrant (Calibrant)
The System is intended to be used by trained operators in a professional laboratory to perform the following key tasks:
- De-salt (using Clean Resin) amplified nucleic acid samples, upon completion of polymerase chain reaction (PCR) and single-base extension reactions following the instructions provided in the Sequenom test-specific package insert;
- Transfer (using the IMPACT Dx NANO) de-salted nucleic acid samples from a microtiter plate onto a disposable 96-pad sample Chip;
- Obtain mass spectra (using the IMPACT Dx MA) from samples and 3-Point Calibrant on a Chip; and
- Analyze (using the TYPER Dx software) the mass spectra of the samples for genotyping results.
The IMPACT Dx System accomplishes genomic analysis and genotyping testing by coupling a biochemistry process (Sequenom biochemistry) that involves target-specific PCR amplification and single-base extension reactions with the subsequent analysis of the reaction products of the target nucleic acids by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This biochemistry process is homogeneous and does not require purification of the PCR products or the extension products and thus is very amenable to high-throughput genotyping testing.
IMPACT Dx NANO
The IMPACT Dx NANO is a self-contained, enclosed instrument that uses computer-controlled robotics to transfer nanoliter volumes of analyte from a 96-well microtiter plate onto a Chip, which is subsequently processed by means of MALDI-TOF MS analysis on the IMPACT Dx MA. This instrument includes an integrated computer pre-loaded with the Nanodispenser software and provides a simple touch-screen interface for users.
IMPACT Dx MA
The IMPACT Dx MA is a bench top mass spectrometer that processes analyte-loaded Chips by means of MALDI-TOF MS analysis. This instrument includes an integrated computer pre-loaded with the TYPER Dx software, a monitor, and a firewall for secure communication with the IMPACT Dx NANO. The main function of the IMPACT Dx MA is to acquire mass spectra from analytes that have been transferred onto a Chip, which has a chemical matrix on each pad. The mass spectra are captured and further analyzed by the TYPER Dx software.
TYPER Dx Software
The TYPER Dx software (TYPER Dx) manages the processing of Sequenom genotyping tests. It is deployed on the computer embedded within the IMPACT Dx MA.
The TYPER Dx software provides the following key functions:
- Allows users to create and manage panel runs;
- Monitors analyte transfer activities on the IMPACT Dx NANO;
- Controls user-initiated, automated mass spectrum acquisition runs on the IMPACT Dx MA;
- Analyzes the mass spectra acquired by the IMPACT Dx MA and makes genotype calls per a test-specific algorithm;
- Enables users to view and export results; and
- Allows an administrator to manage users to ensure secure access to the IMPACT Dx MA and panel run data.
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
trained operators in a professional laboratory
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
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Non-clinical Bench Data:
A series of internal and external analytical studies were conducted which demonstrated that the IMPACT Dx Factor V Leiden and Factor II Genotyping Test has equivalent performance compared to the predicate devices with respect to analytical sensitivity, analytical specificity, reproducibility and potential interfering substances and mutations. In addition, studies were conducted to verify that sample-to-sample contamination did not occur during the test procedure when used with the IMPACT Dx System. Lastly, in a study using fresh EDTA anti-coagulated whole blood samples collected from normal human subjects, DNA was extracted using the recommended DNA extraction method. The IMPACT Dx Factor V Leiden and Factor II Genotyping Test demonstrated 100% concordance with the reference bi-directional sequencing method on genotypes of both Factor II and Factor V genes.
Analytical Specificity:
To determine the specificity of the extension primers were evaluated individually in the extension reactions using 13 clinical genomic DNA from patient samples encompassing all relevant genotypes for Factor VL and Factor II. All clinical samples produced the expected genotype calls, in respect to the extension primer under evaluation. When all extension primers in the IMPACT Dx Factor V Leiden and Factor II Genotyping Test were evaluated together, similarly, all clinical samples produced the expected genotype calls for both FII and FVL.
Interfering Mutations:
In studies to evaluate the effect of mutations in close proximity to genotyping at locus 1691 of the FVL gene, as well as locus 20210 on the FII gene, the variants were tested in each of the 4 reactions. No FVL or FII genotype will be reported on the patient specimens when the Factor V forward assay fails to produce a genotype call due to the presence of an interfering 1690 C>T or 1692A>C or on a patient specimen when the Factor II forward assay fails to produce a genotype call due to the presence of either the 20207A>C or 20209C>T interfering mutations. No miscall or unexpected call from any of the samples was observed during the evaluation.
Analytical Sensitivity:
Clinical genomic DNA obtained from patient samples encompassing all relevant genotypes for Factor V and Factor II genes were used to determine the limits of detection of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test. A series dilution of input DNA levels ranging from 25 ng down to 0.0015 ng was used. The limit of detection was very similar for wild type, heterozygous and homozygous mutant alleles for both loci, and is 0.67 ng per reaction. The recommended input DNA level is 25 ng per reaction.
Carry-over Contamination:
A study to evaluate cross contamination and carry-over was conducted with a panel of 3 clinical genomic DNA samples and a no template control sample. No cross contamination or carry-over was observed as all the no template control samples produced the expected no-call results after the runs.
Interferences:
Leukocyte-depleted whole blood specimens spiked with cell lines from individuals encompassing all relevant genotypes for both the Factor V and Factor II genes were used in this evaluation. The following substances, at the concentrations listed below, were evaluated for their potential interfering effect on the test results.
- Hemoglobin, 200 mg/dL
- K2-EDTA, 3.4 µM
- Heparin, 3000 units/L
- Cholesterol. 500 mg/dL
- Bilirubin, 60 mg/dL
- Ethanol, 500 mg/dL
These substances are representative of metabolites produced during pathological conditions (endogenous) or compounds introduced during sample preparation (exogenous). All the substances were added to whole blood samples before genomic DNA extraction, except ethanol, which was added after the DNA extraction. None of the substances tested adversely impacted the performance of the IMPACT Dx test.
Reproducibility:
The reproducibility of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test was assessed at three external clinical sites. Twelve human genomic DNA samples, including all genotypes (wild type (GG), heterozygous mutant (GA) and homozygous mutant (AA) for both FV and FII genes) were used in this study. Each DNA sample was tested in singlet by each of 3 sites using one discrete IMPACT Dx System at each site. Two operators each conducted 5 runs on 5 non-consecutive days for a total of 10 testing runs per site. The number of correct calls, defined as the number of samples yielding the expected genotypes for both FV and FII genes, no calls and mis-calls were calculated for each operator, and all sites and operators combined. All operators, except one, from all 3 sites produced 100% agreement between the genotypes after all no call results were retested. One operator (Operator 2, Site 1) had 2 samples yielding repeated no calls; there were no mis-calls.
Clinical Data:
In a method comparison study, 860 clinical samples were tested in the IMPACT Dx Factor V Leiden and Factor II Genotyping Test and compared to bi-directional sequencing reference method. The test demonstrated an overall percentage agreement of 99.4% (Cl: 98.6 - 99.8%) and 99.3% (Cl: 98.5 - 99.7%) for the Factor II and Factor V genes, respectively, with the reference method.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
For Factor II: Overall percentage agreement of 99.4% (Cl: 98.6 - 99.8%).
Positive percent agreement (clinical sensitivity) of 100.0% (95% CI: 96.3 – 100.0%).
Negative percent agreement (clinical specificity) of 99.3% (95% Cl: 98.5 - 99.8%).
For Factor V: Overall percentage agreement of 99.3% (Cl: 98.5 - 99.7%).
Positive percent agreement (clinical sensitivity) of 98.7% (95% CI: 95.3 - 99.8%).
Negative percent agreement (clinical specificity) of 99.4% (95% Cl: 98.6 - 99.8%).
Predicate Device(s)
Reference Device(s)
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 864.7280 Factor V Leiden DNA mutation detection systems.
(a)
Identification. Factor V Leiden deoxyribonucleic acid (DNA) mutation detection systems are devices that consist of different reagents and instruments which include polymerase chain reaction (PCR) primers, hybridization matrices, thermal cyclers, imagers, and software packages. The detection of the Factor V Leiden mutation aids in the diagnosis of patients with suspected thrombophilia.(b)
Classification. Class II (special controls). The special control is FDA's guidance entitled “Class II Special Controls Guidance Document: Factor V Leiden DNA Mutation Detection Systems.” (See § 864.1(d) for the availability of this guidance document.)
0
ട്ട് sequenom. _ ............................................................................................................................................................
JUN 1 3 2014
510(K) SUMMARY
This 510(k) Summary is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92
The assigned 510(k) number is: K132978
- A. Submitter Sequenom, Inc. 3595 John Hopkins Court San Diego, California 92121
Contact
Robin Weiner Senior Vice President, Quality and Regulatory. Affairs Sequenom, Inc. Telephone: (858) 202-9044 Telefax: (858) 202-9020 Email: rweiner@sequenom.com
Date Prepared June 12, 2014 .
B. Device Names and Regulatory Information
ASSAY
| Common or Usual Name: | Factor V Leiden and Factor II genotyping test |
|---|---|
| Proprietary Name | IMPACT Dx™ Factor V Leiden and Factor II Genotyping Test |
| Classification Name | Factor V Leiden DNA mutation detection systems (21 CFR |
| 864.7280) | |
| Classification | Class II |
| Product Code | PHJ: System, mass spectrometry, multiplex genotyping, hereditary |
| thrombophilia related mutations | |
| Panel | Hematology (81) |
Pan
C. Predicate Devices
ASSAY :
Roche Factor V Leiden Kit (K033607) Roche Factor II (Prothrombin) G20210A Kit (K033612)
INSTRUMENT
Roche LightCycler® (K033734) AutoGenomics INFINITI™ System (K060564)
IMPACT Dx Factor V Leiden and Factor II Genotyping Test
Confidential Communication Page 1 of 9
1
s sequenom.
D. Device Description
IMPACT Dx Factor V Leiden and Factor II Genotyping Test Overview
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is a qualitative, multiplexed genetic testing device for parallel detection and genotyping of the point mutations G1691A of the Factor V gene and G20210A of the Factor II gene from genomic DNA isolated from EDTA anti- coagulated human whole blood samples. The test is to be performed on the IMPACT Dx System.
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is performed using the IMPACT Dx System, which includes the IMPACT Dx NANO and the IMPACT Dx MA, a matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometer. The test involves Factor V and Factor II region-specific polymerase chain reaction (PCR) amplification of genomic DNA purified from human whole blood in a multiplexed reaction, followed by allele-specific single base primer extension reactions. The reaction products are desalted, dispensed onto a SpectroCHIP® Array using the IMPACT Dx NANO, and the genotyping products are resolved on the basis of mass using the IMPACT Dx MA.
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test provides reagents for multiplex PCR, deoxynucleotide triphosphate dephosphorylation, and single base extension. The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is comprised of the following components:
- . IMPACT Dx Factor V Leiden and Factor II Primer Set
- IMPACT Dx PCR Reagent Set .
- . IMPACT Dx Extend Reagent Set
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test utilizes a biochemistry process (Sequenom Biochemistry) that involves target-specific PCR amplification and single-base extension reactions with the subsequent analysis of the reaction products of the target nucleic acids by matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS),
IMPACT Dx System Overview
The IMPACT Dx System (System) is a platform for highly accurate and sensitive genomic analysis and is designed for use with FDA cleared or approved assays citing its use. The IMPACT Dx System is comprised of the following instruments, software and consumables;
- . IMPACT Dx NANO (NANO)
- IMPACT Dx MA (MA) .
- TYPER Dx Software (TYPER Dx) ●
- System Consumables .
- SpectroCHIP® Arrays (Chip) o
- Clean Resin o
- 3-Point Calibrant (Calibrant) o
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication Page 2 of 9
510(k) Summary
2
sequenom. __
The System is intended to be used by trained operators in a professional laboratory to perform the following key tasks:
- De-salt (using Clean Resin) amplified nucleic acid samples, upon completion of polymerase . chain reaction (PCR) and single-base extension reactions following the instructions provided in the Sequenom test-specific package insert;
- Transfer (using the IMPACT Dx NANO) de-salted nucleic acid samples from a microtiter plate . onto a disposable 96-pad sample Chip;
- . Obtain mass spectra (using the IMPACT Dx MA) from samples and 3-Point Calibrant on a Chip; and
- Analyze (using the TYPER Dx software) the mass spectra of the samples for genotyping results. .
The IMPACT Dx System accomplishes genomic analysis and genotyping testing by coupling a biochemistry process (Sequenom biochemistry) that involves target-specific PCR amplification and single-base extension reactions with the subsequent analysis of the reaction products of the target nucleic acids by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This biochemistry process is homogeneous and does not require purification of the PCR products or the extension products and thus is very amenable to high-throughput genotyping testing.
IMPACT Dx NANO
1
The IMPACT Dx NANO is a self-contained, enclosed instrument that uses computer-controlled robotics to transfer nanoliter volumes of analyte from a 96-well microtiter plate onto a Chip, which is subsequently processed by means of MALDI-TOF MS analysis on the IMPACT Dx MA. This instrument includes an integrated computer pre-loaded with the Nanodispenser software and provides a simple touch-screen interface for users.
IMPACT Dx MA
The IMPACT Dx MA is a bench top mass spectrometer that processes analyte-loaded Chips by means of MALDI-TOF MS analysis. This instrument includes an integrated computer pre-loaded with the TYPER Dx software, a monitor, and a firewall for secure communication with the IMPACT Dx NANO. The main function of the IMPACT Dx MA is to acquire mass spectra from analytes that have been transferred onto a Chip, which has a chemical matrix on each pad. The mass spectra are captured and further analyzed by the TYPER Dx software.
TYPER Dx Software
The TYPER Dx software (TYPER Dx) manages the processing of Sequenom genotyping tests. It is deployed on the computer embedded within the IMPACT Dx MA.
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication Page 3 of 9
510(k) Summary
June 12, 2014
3
sequenom.
The TYPER Dx software provides the following key functions:
- Allows users to create and manage panel runs; �
- Monitors analyte transfer activities on the IMPACT Dx NANO; ●
- Controls user-initiated, automated mass spectrum acquisition runs on the IMPACT Dx MA; .
- Analyzes the mass spectra acquired by the IMPACT Dx MA and makes genotype calls per a test-. specific algorithm;
- Enables users to view and export results; and .
- Allows an administrator to manage users to ensure secure access to the IMPACT Dx MA and panel run data.
E. Intended Use
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is a qualitative in vitro diagnostic device intended for use in the detection and genotyping of a single point mutation (G1691A, referred to as the Factor V Leiden mutation or FVL) of the Factor V gene, located on Chromosome 1q23, and a single point mutation (G20210A) of the prothrombin gene (referred to as Factor II or FII), located on Chromosome 11p11-q12, from genomic DNA isolated from EDTA anti-coagulated human whole blood samples. The test is to be performed on the IMPACT Dx System and is indicated for use as an aid in the diagnosis of patients with suspected thrombophilia.
Comparison with Predicate Devices F.
ASSAY
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is substantially equivalent to the legally marketed predicate devices. These devices have the same intended use and indications for use and similar technological characteristics and principles of operation. They are all PCR-based tests for the genotyping of the Factor V and Factor II genes, with essentially the same performance characteristics. They use different signal detection methodologies and run on different instrumentation platforms; however, all are based on similar technological principles. Table F.1 below presents the similarities and differences between the assay and the predicate devices.
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication Page 4 of 9 .
510(k) Summary
June 12, 2014
4
ള്ള ടലവശ്വാനം.
| Device | Predicate | |||
|---|---|---|---|---|
| IMPACT Dx Factor V | ||||
| Leiden and Factor II | ||||
| Genotyping Test | Roche Factor V Leiden Kit | Roche Factor II | ||
| (Prothrombin) G20210A | ||||
| Kit | ||||
| 510(k) Number | K132978 | K033607 | K033612 | |
| Type of Test | Genotyping Test | Genotyping Test | Genotyping Test | |
| Measurand | Factor II and Factor V | Factor V | Factor II | |
| Target of Detection | Single-nucleotide | |||
| polymorphism | Single-nucleotide | |||
| polymorphism | Single-nucleotide | |||
| polymorphism | ||||
| Intended User | Health Care Professional | Health Care Professional | Health Care Professional | |
| Intended Use | Qualitative in vitro | |||
| diagnostic genotyping test | ||||
| for the detection of Factor II | ||||
| and Factor V alleles from | ||||
| EDTA anti-coagulated | ||||
| human whole blood samples | Qualitative in vitro | |||
| diagnostic genotyping test | ||||
| for the detection Factor V | ||||
| only in EDTA anti- | ||||
| coagulated whole blood | ||||
| samples | Qualitative in vitro | |||
| diagnostic genotyping test | ||||
| for the detection of Factor II | ||||
| only in EDTA anti- | ||||
| coagulated whole blood | ||||
| samples | ||||
| Indications for Use | Aid in the diagnosis of | |||
| patients with suspected | ||||
| thrombophilia | Aid in the diagnosis of | |||
| patients with suspected | ||||
| thrombophilia | Aid in the diagnosis of | |||
| patients with suspected | ||||
| thrombophilia | ||||
| Specimen Type | Purified DNA from human | |||
| blood samples | Purified DNA from human | |||
| blood samples | Purified DNA from human | |||
| blood samples | ||||
| Technological Detection | ||||
| Principles | Genotyping test for | |||
| simultaneous detection | ||||
| (multiplex system) of PCR- | ||||
| amplified DNA sequences | Genotyping test for PCR- | |||
| amplified DNA sequences | Genotyping test for PCR- | |||
| amplified DNA sequences | ||||
| Sample Preparation | DNA extraction and | |||
| purification performed off- | ||||
| line | DNA extraction and | |||
| purification performed off- | ||||
| line | DNA extraction and | |||
| purification performed off- | ||||
| line | ||||
| Oligonucleotide probes and | ||||
| primers | Specific for Factor V Leiden | |||
| (G1691A) and Factor II | ||||
| G20210A | Specific for Factor V Leiden | |||
| (G1691A) | Specific for Factor II | |||
| (prothrombin G20210A) | ||||
| Detection Chemistry | SNP discrimination by | |||
| allele-specific single | ||||
| nucleotide extension coupled | ||||
| with MALDI-TOF mass | ||||
| spectrometry | Fluorogenic detection of | |||
| PCR-amplification products | ||||
| by melting curve analysis | Fluorogenic detection of | |||
| PCR-amplification products | ||||
| by melting curve analysis | ||||
| Analytical Sensitivity | 200 allele copies (0.67 ng | |||
| input DNA/reaction) | FV: 50 allele copies / | |||
| reaction | FII: 50 allele copies / | |||
| reaction | ||||
| Instrument | IMPACT Dx System | Roche LightCycler | Roche LightCycler | |
| Controls | Internal control per sample | |||
| plus external positive and | ||||
| negative controls required | ||||
| per run | External positive and | |||
| negative controls required | ||||
| per run | External positive and | |||
| negative controls required | ||||
| per run | ||||
| Reference Method | Bi-directional DNA | |||
| sequencing | DNA sequencing | DNA sequencing | ||
| Device | Predicate | |||
| Sequenom | ||||
| IMPACT Dx™ System | Roche | |||
| LightCycler® | AutoGenomics | |||
| INFINITI™ System | ||||
| 510(k) Number | K132978 | K033734 | K060564 | |
| Intended Use | Simultaneous, qualitative | |||
| detection of multiple | ||||
| analytes in a PCR amplified | ||||
| genomic DNA sample | ||||
| utilizing MALDI-TOF MS. | A fully automated | |||
| amplification and detection | ||||
| system for nucleic acids | ||||
| using fluorescence | ||||
| detection. | Designed to measure | |||
| fluorescence signals of | ||||
| labeled DNA target | ||||
| hybridized to BioFilmChip | ||||
| microarrays | ||||
| Intended User | Clinical laboratory | Clinical laboratory | Clinical laboratory | |
| Specimen Type | Purified nucleic acids | Purified nucleic acids | Purified nucleic acids | |
| Specimen Preparation | Performed off-line | Performed off-line | Performed off-line | |
| Test Principle | MALDI-TOF detection | |||
| (multiplex system) of PCR- | ||||
| amplified DNA sequences | Fluorogenic detection of | |||
| PCR-amplified DNA | ||||
| fragments by melting curve | ||||
| analysis | Fluorogenic detection | |||
| (multiplex system) of PCR- | ||||
| amplified DNA fragments | ||||
| Detection Procedure | Mass spectrometric analysis | |||
| of target-specific | ||||
| sequences. | Optical detection of | |||
| stimulated fluorescence | Optical detection of | |||
| stimulated fluorescence | ||||
| Detection Chemistry | Single-base extension | |||
| reactions with the | ||||
| subsequent analysis using | ||||
| MALDI-TOF MS | Paired hybridization probes | |||
| using fluorescence | ||||
| resonance energy transfer | Direct fluorescence |
Table F.1 ・ Substantial Equivalence – Comparison to Predicate Devices
:
: - -------------------------------------------------------------------------------------------------------------------------------------------------------------------------
.
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication Page 5 of 9
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sequenom.
INSTRUMENT
The IMPACT Dx System is substantially equivalent to the currently legally marketed predicate devices. These devices have the same intended use and similar technological characteristics and principles of operation. They are PCR-based genotyping instruments using detection of single nucleotide polymorphisms and have essentially the same performance characteristics. Table F.2 below presents the similarities and differences between the IMPACT Dx System and predicate devices.
| Table F.2 | ||||||||
|---|---|---|---|---|---|---|---|---|
| Substantial Equivalence -- Comparison with Predicate Device |
G. Non-clinical Bench Data
A series of internal and external analytical studies were conducted which demonstrated that the IMPACT Dx Factor V Leiden and Factor II Genotyping Test has equivalent performance compared to the predicate devices with respect to analytical sensitivity, analytical specificity, reproducibility and potential interfering substances and mutations. In addition, studies were conducted to verify that sample-to-sample contamination did not occur during the test procedure when used with the IMPACT Dx System. Lastly, in a study using fresh EDTA anti-coagulated whole blood samples collected from normal human subjects, DNA was extracted using the recommended DNA extraction method. The IMPACT Dx Factor V Leiden and Factor II Genotyping Test demonstrated 100% concordance with the reference bi-directional sequencing method on genotypes of both Factor II and Factor V genes.
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication Page 6 of 9
June 12, 2014
6
sequenom.
Analytical Specificity
To determine the specificity of the extension primers were evaluated individually in the extension reactions using 13 clinical genomic DNA from patient samples encompassing all relevant genotypes for Factor VL and Factor II. All clinical samples produced the expected genotype calls, in respect to the extension primer under evaluation. When all extension primers in the IMPACT Dx Factor V Leiden and Factor II Genotyping Test were evaluated together, similarly, all clinical samples produced the expected genotype calls for both FII and FVL.
Interfering Mutations
In studies to evaluate the effect of mutations in close proximity to genotyping at locus 1691 of the FVL gene, as well as locus 20210 on the FII gene, the variants were tested in each of the 4 reactions. No FVL or FII genotype will be reported on the patient specimens when the Factor V forward assay fails to produce a genotype call due to the presence of an interfering 1690 C>T or 1692A>C or on a patient specimen when the Factor II forward assay fails to produce a genotype call due to the presence of either the 20207A>C or 20209C>T interfering mutations. No miscall or unexpected call from any of the samples was observed during the evaluation.
Analytical Sensitivity
Clinical genomic DNA obtained from patient samples encompassing all relevant genotypes for Factor V and Factor II genes were used to determine the limits of detection of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test. A series dilution of input DNA levels ranging from 25 ng down to 0.0015 ng was used. The limit of detection was very similar for wild type, heterozygous and homozygous mutant alleles for both loci, and is 0.67 ng per reaction. The recommended input DNA level is 25 ng per reaction.
Carry-over Contamination
A study to evaluate cross contamination and carry-over was conducted with a panel of 3 clinical genomic DNA samples and a no template control sample. No cross contamination or carry-over was observed as all the no template control samples produced the expected no-call results after the runs.
Interferences
Leukocyte-depleted whole blood specimens spiked with cell lines from individuals encompassing all relevant genotypes for both the Factor V and Factor II genes were used in this evaluation. The following substances, at the concentrations listed below, were evaluated for their potential interfering effect on the test results.
- Hemoglobin, 200 mg/dL .
- K2-EDTA, 3.4 µM ●
- . Heparin, 3000 units/L
- . Cholesterol. 500 mg/dL
- Bilirubin, 60 mg/dL .
- Ethanol, 500 mg/dL
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication
510(k) Summary
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These substances are representative of metabolites produced during pathological conditions (endogenous) or compounds introduced during sample preparation (exogenous). All the substances were added to whole blood samples before genomic DNA extraction, except ethanol, which was added after the DNA extraction. None of the substances tested adversely impacted the performance of the IMPACT Dx test.
Reproducibility
The reproducibility of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test was assessed at three external clinical sites. Twelve human genomic DNA samples, including all genotypes (wild type (GG), heterozygous mutant (GA) and homozygous mutant (AA) for both FV and FII genes) were used in this study. Each DNA sample was tested in singlet by each of 3 sites using one discrete IMPACT Dx System at each site. Two operators each conducted 5 runs on 5 non-consecutive days for a total of 10 testing runs per site. The number of correct calls, defined as the number of samples yielding the expected genotypes for both FV and FII genes, no calls and mis-calls were calculated for each operator, and all sites and operators combined. All operators, except one, from all 3 sites produced 100% agreement between the genotypes after all no call results were retested. One operator (Operator 2, Site 1) had 2 samples vielding repeated no calls; there were no mis-calls.
H. Clinical Data
In a method comparison study, 860 clinical samples were tested in the IMPACT Dx Factor V Leiden and Factor II Genotyping Test and compared to bi-directional sequencing reference method. Table H.1 presents the percent agreement between the IMPACT Dx Factor V Leiden and Factor II Genotyping Test and the reference method along with the respective lower confidence boundary (LCB) of the 95% confidence interval (CI) for Factor II. Table H.2 presents similar information for Factor V.
The test demonstrated an overall percentage agreement of 99.4% (Cl: 98.6 - 99.8%) and 99.3% (Cl: 98.5 - 99.7%) for the Factor II and Factor V genes, respectively, with the reference method.
| Genotype
By
Sequencing | #
Samples | Number of FII Calls Before Repeat Testing | | | | Number of FII Calls After Repeat Testing | | | | | | |
|------------------------------|--------------|-------------------------------------------|----------------|-----------------------|----------------|------------------------------------------|------------------|----------------|-----------------------|----------------------|----------------|-------------|
| | | Correct
Calls | # No
Calls¹ | #
Missed
Calls² | %
Agreement | 95%
LCB³ | Correct
Calls | # No
Calls¹ | #
Missed
Calls² | # Repeat
Samples⁴ | %
Agreement | 95%
LCB³ |
| Wild Type | 762 | 747 | 15 | 0 | 98.0 | 97.0 | 757 | 5 | 0 | 15 | 99.3 | 98.5 |
| Heterozygous | 78 | 77 | 1 | 0 | 98.7 | 94.1 | 78 | 0 | 0 | 1 | 100.0 | 97.1 |
| Homozygous | 19 | 19 | 0 | 0 | 100.0 | 88.6 | 19 | 0 | 0 | 0 | 100.0 | 88.6 |
| All Samples | 859 | 843 | 16 | 0 | 98.1 | 97.0 | 854 | 5 | 0 | 16 | 99.4 | 98.6 |
Table H.1 Comparison of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test and DNA Sequencing Results - Factor II
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| Table H.2 |
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| Comparison of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test and DNA Sequencing |
| Results – Factor V |
| Genotype
By
Sequencing | #
Samples | Number of FVL Calls Before Repeat Testing | | | | | Number of FVL Calls After Repeat Testing | | | | | |
|------------------------------|--------------|-------------------------------------------|----------------|-----------------------|----------------|-------------|------------------------------------------|----------------|-----------------------|----------------------|----------------|-------------|
| | | Correct
Calls | # No
Calls1 | #
Missed
Calls2 | %
Agreement | 95%
LCB3 | Correct
Calls | # No
Calls1 | #
Missed
Calls2 | # Repeat
Samples4 | %
Agreement | 95%
LCB3 |
| Wild Type | 710 | 697 | 13 | 0 | 98.2 | 97.1 | 706 | 4 | 0 | 13 | 99.4 | 98.6 |
| Heterozygous | 132 | 127 | 5 | 0 | 96.2 | 92.2 | 130 | 2 | 0 | 5 | 98.5 | 95.3 |
| Homozygous | 18 | 18 | 0 | 0 | 100.0 | 88.0 | 18 | 0 | 0 | 0 | 100.0 | 88.0 |
| All Samples | 860 | 842 | 18 | 0 | 97.9 | 96.7 | 854 | 6 | 0 | 18 | 99.3 | 98.5 |
When the heterozygous and homozygous mutant genotypes are combined into one "Positive" category and compared to one "Negative" (wild type) category, the test demonstrated a positive percent agreement or clinical sensitivity of 100.0% (95% CI: 96.3 – 100.0%) and a negative percent agreement or clinical specificity of 99.3% (95% Cl: 98.5 - 99.8%) for Factor II. Similarly, for Factor V, positive percent agreement or clinical sensitivity was 98.7% (95% CI: 95.3 - 99.8%), and negative percent agreement or clinical specificity was 99.4% (95% Cl: 98.6 - 99.8%) for Factor V.
I. Conclusion
The intended use of the IMPACT Dx Factor V Leiden and Factor II Genotyping Test is to measure a single point mutation (G1691A, referred to as the Factor V Leiden mutation or FVL) of the Factor V gene, located on Chromosome 1q23, and a single point mutation (G20210A) of the prothrombin gene (referred to as Factor II or F11), located on Chromosome 11p11-q12, from genomic DNA isolated from EDTA anti-coagulated human whole blood samples as an aid in the diagnosis of patients with suspected thrombophilia.
Performance data demonstrate that the IMPACT Dx Factor V Leiden and Factor II Genotyping Test using the IMPACT Dx System is as safe and effective as predicate devices. Thus the IMPACT Dx System and the IMPACT Dx Factor V Leiden and Factor II Genotyping Test are substantially equivalent to the other legally marketed devices, supporting premarket 510(k) clearance,
IMPACT Dx Factor V Leiden and Factor II Genotyping Test K132978 Confidential Communication Page 9 of 9
510(k) Summary
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Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
June 13, 2014
Sequenom, Inc. Ms. Robin Weiner Senior Vice President, Quality and Regulatory Affairs 3595 John Hopkins Court San Diego, CA 92121
Re: K132978
Trade/Device Name: IMPACT Dx™ Factor V Leiden and Factor II Genotyping Test on the IMPACT Dx™ System Regulation Number: 21 CFR 864.7280 Regulation Name: Factor V Leiden DNA Mutation Detection Systems Regulatory Class: Class II Product Code: PHJ Dated: May 13, 2014 Received: May 14, 2014
Dear Ms. Weiner:
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. Please note: CDRH does not evaluate information related to contract liability warranties. 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 subject to additional 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 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 Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set
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forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.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 CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours.
' Reena Philip -S
Reena Philip, Ph.D. Director Division of Molecular Genetics and Pathology Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
11
DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration
Indications for Use
510(k) Number (if known) K132978
Device Name
IMPACT Dx™ Factor V Leiden and Factor II Genotyping Test on the IMPACT™ Dx System
Indications for Use (Describe)
The IMPACT Dx Factor V Leiden and Factor II Genotyping Test is a qualitative in vitro diagnostic device intended for use in the detection and genotyping of a single point mutation (G1691A, referred to as the Factor V Leiden mutation or FVL) of the Factor V gene, located on Chromosome 1q23, and a single point mutation (G20210A) of the probrombin gene (referred to as Factor II or FII), located on Chromosome 11p11-q12, from genomic DNA isolated from EDTA anticoagulated human whole blood samples. The test is to be performed on the IMPACT Dx System and is indicated for use as an aid in the diagnosis of patients with suspected thrombophilia.
Type of Use (Select one or both, as applicable)
2 Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Donna M. Roscoe -S
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