(582 days)
The Vysis CLL FISH Probe Kit is intended to detect deletion of the LS1 TP53, LSI ATM, and LSI D13S319 probe targets and gain of the D12Z3 sequence in peripheral blood specimens from untreated patients with B-cell chronic lymphocytic leukemia (CLL). The assay may be used to dichotomize CLL (the 13q-, +12, or normal genotype group versus the 11q- or 17p- group) and may be used as an aid in determining disease prognosis in combination with additional biomarkers, morphology, and other clinical information. The Vysis CLL FISH Probe Kit is not intended for use in selection of therapy or in monitoring of residual disease.
The Vysis CLL FISH Probe Kit uses fluorescence in situ hybridization (FISH) DNA probe technology to determine deletions of the locus-specific identifier (LSI) TP53, LSI ATM, and LSI D13S319 probe targets and gain of the D12Z3 sequence.
The Vysis CLL FISH Probe Kit (List No. 4N02-020) consists of two DNA FISH probe sets and three general purpose reagents sufficient to process 20 assays.
- . LSI TP53 SpectrumOrange/ATM SpectrumGreen Probe
- LSI D13S319 SpectrumOrange/13q34 SpectrumAqua/CEP 12 SpectrumGreen Probe .
- DAPI II Counterstain .
- NP-40 .
- 20X SSC Salt .
Here's a summary of the acceptance criteria and the study details for the Vysis CLL FISH Probe Kit, based on the provided 510(k) summary:
Acceptance Criteria and Device Performance
Note: The 510(k) summary for the Vysis CLL FISH Probe Kit primarily focuses on analytical performance (specificity, sensitivity, normal cut-off values, precision, reproducibility) and concordance with an existing clinical assay rather than traditional "acceptance criteria" related to a new AI/CADe device. The provided tables outline the device's technical performance.
| Acceptance Criterion / Performance Metric | Reported Device Performance (Mean/Overall) |
|---|---|
| Analytical Specificity | 100% for all probes (LSI TP53, LSI ATM, LSI D13S319, LSI 13q34, CEP 12) |
| Analytical Sensitivity (Expected normal signal pattern: 2 signals/nucleus) | LSI TP53: 97.98% |
| LSI ATM: 98.68% | |
| LSI D13S319: 98.60% | |
| CEP 12: 98.94% | |
| Normal Cut-off Values (Percentage of abnormal nuclear FISH patterns) | LSI TP53 (1 signal): 7.0% (14/200 nuclei) |
| LSI ATM (1 signal): 6.0% (12/200 nuclei) | |
| LSI D13S319 (1 signal): 5.5% (11/200 nuclei) | |
| LSI D13S319 (0 signal): 1.5% (3/200 nuclei) | |
| CEP 12 (3 signals): 2.5% (5/200 nuclei) | |
| Precision (Mean % abnormal cells for various probes/abnormalities) | Ranges from 0.0% to 73.2% across different samples and probes (SDs provided in Tables 4-8) |
| Reproducibility (Overall Agreement, Site to Site by Probe) | TP53 (17p-): 100% |
| ATM (11q-): 90% | |
| CEP 12 (+12): 100% | |
| D13S319 1x (13q-): 90% | |
| D13S319 2x (13q-): 90% | |
| Reproducibility (Prognostic Category, Generalized Kappa) | Kappa = 0.86 (Strength: "Almost Perfect") |
| Method Concordance (AMT vs. RFT for Prognostic Category) | Overall Agreement: 97% (62/64) |
| Lower bound one-sided 95% CI: 90% |
Study Details
-
Sample sizes used for the test set and the data provenance:
- Analytical Specificity: 5 karyotypically normal male peripheral blood cultures (retrospective, assumed domestic, not explicitly stated).
- Analytical Sensitivity: 25 karyotypically normal patient peripheral blood cultures (retrospective, assumed domestic, not explicitly stated).
- Analytical Characterization of Normal Cut-off Values: 25 karyotypically normal patient peripheral blood cultures (retrospective, assumed domestic, not explicitly stated).
- Precision:
- Precision Study 1: 2 negative peripheral blood specimens (lacking abnormalities) and 8 additional specimens with at least one abnormality. Likely retrospective, but not explicitly stated.
- Precision Study 2: 8 different patient specimens (blinded panel). Likely retrospective, but not explicitly stated.
- Reproducibility: A blinded 20-member slide panel representing five Döhner classifications. Likely retrospective, but not explicitly stated.
- Method Concordance: 64 specimens with pre-established Döhner classifications. Retrospective, as classifications were "based on previous results using the RFT." Provenance not explicitly stated.
-
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- Analytical Specificity: One technologist. Qualifications not specified beyond being a "technologist."
- Analytical Sensitivity: Not explicitly stated, but each technologist evaluated 100 nuclei per specimen, implying multiple technologists were involved. Qualifications not specified.
- Analytical Characterization of Normal Cut-off Values: Not explicitly stated, but implies multiple technologists based on the sensitivity study. Qualifications not specified.
- Precision: Not specified, but involved counting signals, implying trained technologists.
- Reproducibility: Not specified, but involved three different laboratories, implying multiple trained personnel.
- Method Concordance: The "Reference FISH Test (RFT)" was used to establish initial Döhner classifications. The original Shanafelt study (referred to as the RFT) implies multiple pathologists/cytogeneticists. This study used "previous results" from the RFT as its ground truth.
-
Adjudication method for the test set:
- No formal adjudication method (like 2+1 or 3+1) is explicitly described for establishing the ground truth or validating results in the analytical studies. Results were typically enumerated by technologists and aggregated.
- For the Reproducibility study, "overall agreement" between three testing sites was assessed, but a specific adjudication process for discrepancies is not detailed beyond reporting disagreement counts.
- For Method Concordance, the ground truth was the "Reference FISH Test (RFT) used in the Shanafelt study," which effectively served as the gold standard, and the AMT results were compared against it.
-
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No MRMC comparative effectiveness study involving human readers improving with AI vs. without AI assistance was reported. This device is a FISH probe kit, not an AI-powered diagnostic system. The studies focused on the analytical and clinical validity of the probe kit itself.
-
If a standalone (i.e., algorithm only without human-in-the-loop performance) was done:
- This is not an algorithm-only device. It is a laboratory assay (FISH probe kit) where human technologists perform the analysis by counting fluorescent signals under a microscope. The studies performed were of the laboratory kit's performance, not an automated algorithm.
-
The type of ground truth used:
- Analytical Specificity, Sensitivity, Normal Cut-off, Precision: Based on visual identification and enumeration of FISH signals by trained technologists, comparing observed signal patterns to expected normal or abnormal patterns in karyotypically normal and patient samples. The "expected normal" pattern serves as a form of expert-derived ground truth.
- Reproducibility: Comparison of results across multiple labs/readers for the same samples. The "agreement" between sites implies a consensus or majority rule for comparison, but the ultimate ground truth for a given sample's true Döhner classification is not explicitly detailed but likely derived from expert pathological/cytogenetic review.
- Method Concordance: The ground truth was established by a "Reference FISH Test (RFT) used in the Shanafelt study," which is stated to have its clinical validity documented via peer-reviewed literature. This implies a ground truth based on established clinical and pathological diagnosis as determined by a validated method.
-
The sample size for the training set:
- This device is a diagnostic kit, not an AI/machine learning algorithm, so there is no "training set" in the conventional sense of AI development. The studies described are for analytical validation and clinical concordance.
-
How the ground truth for the training set was established:
- Since there's no AI "training set," this question is not applicable. The device's performance characteristics (specificity, sensitivity, cut-offs) are established through testing on defined sets of samples (e.g., karyotypically normal individuals, patients with confirmed CLL aberrations) where the expected outcome is known or determined by expert review using established cytogenetic methods.
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510(k) Summary
510(k) Number: K100015
Device Name: Vysis CLL FISH Probe Kit
Purpose of the Submission
The purpose of this 510(k) is to gain clearance to market the Vysis CLL FISH Probe Kit (List No. 4N02-020).
Official Correspondent to the File
| Name: | Dr. Nancy Bengtson |
|---|---|
| Title: | Manager, Regulatory Affairs |
| Telephone: | (224) 361-7087 |
| Fax: | (847) 775-6777 |
| Email: | Nancy.Bengtson@abbott.com |
| Name: | Ms. Pamela L. Swatkowski |
| Title: | Director of Regulatory Affairs |
| Telephone: | (224) 361-7013 |
| Fax: | (847) 775-6777 |
| Email: | Pamela.Swatkowski@abbott.com |
| Address: | Abbott Molecular Inc. |
| 1300 E. Touhy Avenue | |
| Des Plaines. IL 60018 |
Date of Preparation
August 9, 2011
Manufacturer
Abbott Molecular Inc. is the legal manufacturer of the Vysis CLL FISH Probe Kit (List
No. 4N02-020).
| Name: | Timothy Zurow, PhD |
|---|---|
| Title: | Director Manufacturing Operations |
| Telephone: | (224) 361-7379 |
| Fax: | (224) 361-7438 |
| Email: | timothy.zurow@abbott.com |
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
510(k) Summary Page 1 of 20
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| Address: | Abbott Molecular Inc.1300 E. Touhy AvenueDes Plaines, IL 60018 |
|---|---|
| ---------- | ------------------------------------------------------------------------ |
Establishment Registration No.: 3005248192
Intended Use
The Vysis CLL FISH Probe Kit is intended to detect deletion of the LS1 TP53, LSI ATM, and LSI D13S319 probe targets and gain of the D12Z3 sequence in peripheral blood specimens from untreated patients with B-cell chronic lymphocytic leukemia (CLL). The assay may be used to dichotomize CLL (the 13q-, +12, or normal genotype group versus the 11q- or 17p- group) and may be used as an aid in determining disease prognosis in combination with additional biomarkers, morphology, and other clinical information. The Vysis CLL FISH Probe Kit is not intended for use in selection of therapy or in monitoring of residual disease.
Trade Name
Vysis CLL FISH Probe Kit
Common Name
Fluorescence In Situ Hybridization (FISH) reagents
Classification
Class II
Regulation Number
21 CFR 866.6040 Gene expression profiling test system for breast cancer prognosis
Product Code
OVQ, Chronic Lymphocytic Leukemia FISH Probe Kit
Predicate Devices
Agendia BV MammaPrint® (K062694)
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
510(k) Summary Page 2 of 20
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Comparison with Predicate
Not applicable. Clearance is supported by published clinical study and method concordance bridging studies.
Device Description
The Vysis CLL FISH Probe Kit uses fluorescence in situ hybridization (FISH) DNA probe technology to determine deletions of the locus-specific identifier (LSI) TP53, LSI ATM, and LSI D13S319 probe targets and gain of the D12Z3 sequence.
The Vysis CLL FISH Probe Kit (List No. 4N02-020) consists of two DNA FISH probe sets and three general purpose reagents sufficient to process 20 assays.
- . LSI TP53 SpectrumOrange/ATM SpectrumGreen Probe
- LSI D13S319 SpectrumOrange/13q34 SpectrumAqua/CEP 12 SpectrumGreen Probe .
- DAPI II Counterstain .
- NP-40 .
- 20X SSC Salt .
Background on Chronic Lymphocytic Leukemia (CLL)
Currently, most patients diagnosed with CLL have early-stage disease (Rai stage 0 or 1). Patients with early-stage CLL are a heterogeneous group; approximately 30% to 50% are at high risk of accelerated disease progression, and the remainder may live for decades and possibly never require therapy. Recent insights into the biological characteristics of leukemic B cells have led to the discovery of new prognostic tools (immunoglobulin variable-region heavy chain gene mutation status, cytogenetic abnormalities assessed by FISH, and Z-chain-associated protein kinase-70 protein expression) that can contribute to the identification of patients with early-stage disease who are at high risk for early disease progression. '
Routine karyotype analysis only detects chromosomal aberrations associated with CLL in 40% to 50% of the cases. Use of FISH and other technologies have detected genomic
510(k) Summary Page 3 of 20
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abnormalities in over 80% of cases of CLL. The common genomic aberrations seen are trisomy 12 and deletions of 13q, 17p, and 11q.2-4
Several published studies suggest that some of these chromosomal abnormalities may be correlated with various disease parameters.5-8
The Vysis CLL FISH Probe Kit uses FISH DNA probe technology to determine deletion status of probe targets for LSI TP53 (containing tumor protein p53 gene, located on chromosome 17p), LSI ATM (containing ataxia telangjectasia mutated gene, located on chromosome 11q), and LSI D13S319 (containing marker D13S319, located on chromosome 13q), as well as determining trisomy 12 with CEP12 (D12Z3 alpha satellitelocation chromosome 12).
The Vysis CLL FISH Probe Kit includes LSI 13q34 (containing lysosomal-associated membrane protein I gene, located on chromosome 13g) as a quality control probe.
Technological Description of the Device
FISH is a technique that allows visualization of specific nucleic acid sequences within a cellular preparation. Specifically, FISH involves precise annealing of a single-stranded, fluorophore-labeled DNA probe to a complementary target sequence. Hybridization of the probe with the cellular DNA site is visible by direct detection using fluorescence microscopy. Interpretation of FISH results should be made utilizing appropriate controls and analytical techniques as well as taking into consideration other clinical and diagnostic test data.9
Peripheral blood cells from CLL patients are attached to microscope slides using standard cytogenetic procedures. The resulting specimen DNA is denatured to single-stranded form and subsequently allowed to hybridize with the probes of the CLL FISH Probe Kit. Following hybridization, the unbound probe is removed by a series of washes, and the nuclei are counter-stained with DAPI, a DNA-specific stain that fluoresces blue, Hybridization of the Vysis LSI TP53 SpectrumOrange, LSI ATM SpectrumGreen, LSI D13S319 SpectrumOrange, LSI 13q34 SpectrumAqua, and CEP 12 SpectrumGreen probes is viewed using a fluorescence microscope equipped with appropriate excitation
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
510(k) Summary Page 4 of 20
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and emission filters, allowing visualization of the orange, green, and aqua fluorescent signals.
In a cell with normal copy numbers of the LSI TP53 SpectrumOrange and LSI ATM SpectrumGreen probe targets, two orange and two green signals will be expected. In a cell with normal copy numbers of Vysis LSI D13S319 SpectrumOrange and CEP 12 SpectrumGreen probe targets, two orange signals and two green signals will be expected. Enumeration of the Vysis LSI TP53 SpectrumOrange, LSI ATM SpectrumGreen, LSI D13S319 SpectrumOrange, and CEP12 SpectrumGreen signals provides a mechanism for determining absolute copy number of the probe targets and the presence of the aberrations of interest. Any aberrations detected are used to determine Döhner Classification and prognostic category.
Summary of Nonclinical Studies
Analytical Specificity
Analytical specificity is defined as the percentage of signals that hybridize to the correct locus and no other location. This test is a visual demonstration that each probe hybridizes specifically to the expected chromosome location. The analytical specificity of the probes in the Vysis CLL FISH Probe Kit for their respective chromosome target loci was established using metaphase chromosomes prepared from peripheral blood cultures of five karyotypically normal males on microscope slides. The hybridization location of each FISH signal on chromosomes of 20 consecutive metaphase nuclei on each of 5 slides was evaluated by one technologist for a total of 200 target loci.
For each probe and sample, the number of metaphase chromosome FISH signals hybridized to the correct locus and the number of metaphase chromosome FISH signals hybridized to the incorrect locus were enumerated. The specificity of each probe was calculated as the number of metaphase chromosome FISH signals hybridized to the correct locus divided by the total number of metaphase chromosome FISH signals hybridized and multiplied by 100 to give a percentage. The analytical specificity for each probe of the Vysis CLL FISH Probe Kit was 100% (200/200) (Table 1).
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| No. of MetaphaseChromosome Signals | |||||
|---|---|---|---|---|---|
| Probe | CorrectCytogeneticTarget | Hybridized tothe CorrectTarget Locus | TotalHybridizedSignals | Specificity(%) | 95%ConfidenceInterval (%) |
| Vysis LSI TP53SpectrumOrange | 17p13.1 | 200 | 200 | 100 | 98.17, 100 |
| Vysis LSI ATMSpectrumGreen | 11q22.3 | 200 | 200 | 100 | 98.17, 100 |
| Vysis LSID13S319SpectrumOrange | 13q14.3 | 200 | 200 | 100 | 98.17, 100 |
| Vysis LSI 13q34SpectrumAqua | 13q34 | 200 | 200 | 100 | 98.17, 100 |
| Vysis CEP 12SpectrumGreen | 12p11.1-q11 | 200 | 200 | 100 | 98.17, 100 |
Table 1. Analytical Specificity of Probes in Vysis CLL FISH Probe Kit
Analytical Sensitivity
Analytical sensitivity is defined as the percentage of scoreable interphase nuclei with the expected normal signal pattern. The expected normal interphase signal pattern for all probes in the Vysis CLL FISH Probe Kit is two signals per nucleus.
The analytical sensitivity of the probes in the Vysis CLL FISH Probe Kit for their respective chromosome target loci was established using interphase nuclei prepared from peripheral blood cultures of 25 karyotypically normal patients. For each specimen, the signal patterns of 200 nuclei were evaluated by counting the number of orange and green signals present for each probe target. Each technologist evaluated 100 nuclei per specimen for a total of 200 nuclei per specimen and 5000 scoreable nuclei from normal specimens.
The sensitivity (with 95% confidence intervals based on binomial distribution) was calculated as the percentage of scoreable interphase nuclei with the expected signal pattern of two signals per nucleus.
The Vysis CLL FISH Probe Kit has a sensitivity of 97.98% for the LSI TP53 SpectrumOrange probe, 98.68% for the LSI ATM SpectrumGreen probe, 98.60% for the LSI D13S319 SpectrumOrange probe, and 98.94% for the CEP 12 SpectrumGreen probe (Table 2).
510(k) Summary Page 6 of 20
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| Number of Interphase Nuclei | Sensitivity | |||
|---|---|---|---|---|
| Probe | With ExpectedSignal Pattern | ScoreableSignals | Percent (%) | 95%ConfidenceInterval(%) |
| Vysis LSI TP53 SpectrumOrange | 4899 | 5000 | 97.98 | 97.55, 98.35 |
| Vysis LSI ATM SpectrumGreen | 4934 | 5000 | 98.68 | 98.32, 98.98 |
| Vysis LSI D13S319SpectrumOrange | 4930 | 5000 | 98.60 | 98.23, 98.91 |
| Vysis CEP 12 SpectrumGreen | 4947 | 5000 | 98.94 | 98.62, 99.21 |
Table 2. Analytical Sensitivity and Scoreable Percentage for Each Probe in the Vysis CLL FISH Probe Kit
Analytical Characterization of Normal Cut-off Values
The normal cut-off value, in association with FISH DNA probes, is defined as the maximum percentage of scoreable interphase nuclei with a specific abnormal signal pattern at which a specimen is considered normal for that signal pattern. The normal cutoff value is expressed in terms of a percentage or the actual number of abnormal nuclear FISH patterns per the standard number of nuclei tested. The criteria used to classify a nucleus as scoreable is located in the Interpretation and Result Reporting section of the package insert. The Quality Control and Signal Enumeration sections and the Dual Color Signal Counting Guide instruct the end user to determine whether an entire slide is adequate for signal enumeration and which types of cells/signals can be enumerated.
The normal cut-off values of the probes in the Vysis CLL FISH Probe Kit for their respective chromosome target loci were established using interphase nuclei prepared from peripheral blood cultures of 25 karyotypically normal patients. For each specimen, the signal patterns of 200 nuclei were evaluated by counting the number of orange and green signals present for each probe target.
The expected normal interphase signal pattern for all probes in the Vysis CLL FISH Probe Kit was two signals per nucleus. Since the specimen population does not fit a Gaussian distribution, the normal cut-off value was calculated using the beta inverse function. 9
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The Vysis CLL FISH Probe Kit was shown to have normal cut-off values of 14 out of 200 nuclei evaluated (7.0%) for the LSI TP53 SpectrumOrange probe (1 signal), 12 out of 200 nuclei evaluated (6.0%) for the LSI ATM SpectrumGreen probe (1 signal), 11 out of 200 nuclei evaluated (5.5%) for the LSI D13S319 SpectrumOrange probe (1 signal), 3 out of 200 nuclei evaluated (1.5%) for the LSI D13S319 SpectrumOrange probe (0 signal), and 5 out of 200 nuclei evaluated (2.5%) for the CEP 12 SpectrumGreen probe (3 signals) (Table 3).
| Probe(Abnormal Signal Patternof Interest) | Number ofNucleiEvaluated (n) | MaximumNumber of False-Positive Patterns | NormalCut-off Value(per 200 nuclei) | Normal Cut-off Value(%) |
|---|---|---|---|---|
| Vysis LSI TP53SpectrumOrange (1 signal) | 200 | 8 | 14 | 7.0 (14/200) |
| Vysis LSI ATMSpectrumGreen (1 signal) | 200 | 6 | 12 | 6.0 (12/200) |
| Vysis LSI D13S319SpectrumOrange (1 signal) | 200 | 5 | 11 | 5.5 (11/200) |
| Vysis LSI D13S319SpectrumOrange (0 signal) | 200 | 0 | 3 | 1.5 (3/200) |
| Vysis CEP 12SpectrumGreen (3 signals) | 200 | 1 | 5 | 2.5 (5/200) |
Table 3. Analytical Characterization of Normal Cut-off Values
Precision
The precision of the probes in the Vysis CLL FISH Probe Kit was established using interphase nuclei prepared from two separate peripheral blood specimens lacking del(17p13.1), del(11q22.3), del(13q14.3), and trisomy 12, and eight additional specimens of which at least two specimens had one of the listed abnormalities (Precision Study 1).
This blinded 10-member slide panel consisting of both negative and positive specimens was used to test three lots of Vysis LSI TP53 SpectrumOrange/ATM SpectrumGreen and LSI D13S319 SpectrumOrange/13q34 SpectrumAqua/CEP 12 SpectrumGreen Probes on each of two days, and one of the three lots on a third day.
For each specimen, the FISH signal patterns of 200 nuclei were evaluated by counting the number of orange and green signals present for each probe target.
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Precision was analyzed separately for each specimen per probe using the percentage of abnormal cells with the signal pattern(s) of interest.
A blinded panel created from eight different patient specimens was tested using three different DNA FISH probe lots on three days (the days were not required to be consecutive) (Precision Study 2). Precision analyses to challenge the normal cut-off values were performed using the percentage of cells with the signal pattern of interest and were analyzed separately for each panel member.
Precision Study 1 and Precision Study 2 mean and standard deviations of the observed percentages of abnormal cells of the negative, positive, and specimens near the normal cut-off are shown in Tables 4-8.
| Study | Sample | Category | n | Mean | Between-Day(Within Lot)Component SD | Between-LotComponentSD | Total SDb |
|---|---|---|---|---|---|---|---|
| 1 | 1 | Negative | 7 | 2.6 | 1.23 | 0.00 | 1.23 |
| 1 | 2 | Negative | 7 | 2.9 | 1.58 | 1.11 | 1.94 |
| 1 | 3 | Negative | 7 | 3.8 | 3.48 | 0.00 | 3.48 |
| 1 | 4 | Negative | 7 | 3.1 | 2.00 | 0.00 | 2.00 |
| 1 | 5 | Negative | 7 | 2.7 | 0.76 | 0.65 | 1.00 |
| 1 | 6 | Negative | 7 | 2.4 | 2.90 | 0.00 | 2.90 |
| 1 | 7 | Negative | 7 | 2.1 | 1.38 | 0.00 | 1.38 |
| 1 | 8 | Negative | 7 | 2.8 | 1.29 | 0.00 | 1.29 |
| 1 | 9 | Positive | 7 | 29.8 | 5.15 | 4.54 | 6.87 |
| 1 | 10 | Positive | 7 | 73.2 | 5.29 | 0.00 | 5.29 |
| 2 | 7 | Positivec | 9 | 13.6 | 3.11 | 0.00 | 3.11 |
| 2 | 8 | Positivec | 9 | 16.9 | 3.90 | 0.00 | 3.90 |
Table 4. Precision Analysis of Percentages of Abnormal Signal Patterns for Vysis LSI TP53 SpectrumOrange Probe [del(17q13.1(1 signal))]4
The mean and standard deviations are represented as percent abnormal signal patterns.
6 Total variance is the sum of the other variance components.
& Positive specimen near the normal cut-off.
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| Study | Sample | Category | n | Mean | Between-Day(Within Lot)Component SD | Between-LotComponentSD | Total SDb |
|---|---|---|---|---|---|---|---|
| 1 | 1 | Negative | 7 | 1.6 | 0.54 | 0.59 | 0.80 |
| 1 | 2 | Negative | 7 | 1.8 | 0.59 | 0.00 | 0.59 |
| 1 | 3 | Negative | 7 | 2.6 | 0.81 | 1.54 | 1.74 |
| 1 | 4 | Negative | 7 | 2.1 | 2.25 | 1.08 | 2.50 |
| 1 | 5 | Negative | 7 | 2.9 | 1.71 | 1.64 | 2.37 |
| 1 | 6 | Negative | 7 | 1.1 | 1.34 | 0.00 | 1.34 |
| 1 | 7 | Positive | 7 | 64.1 | 3.18 | 0.00 | 3.18 |
| 1 | 8 | Positive | 7 | 13.7 | 4.31 | 5.10 | 6.68 |
| 1 | 9 | Negative | 7 | 1.6 | 0.99 | 0.00 | 0.99 |
| 1 | 10 | Negative | 7 | 1.5 | 1.51 | 0.00 | 1.51 |
| 2 | 1 | Positivec | 9 | 8.4 | 3.12 | 0.91 | 3.25 |
| 2 | 2 | Positivec | 9 | 7.8 | 3.00 | 1.89 | 3.55 |
| 2 | 8 | Positivec | 9 | 18.7 | 3.46 | 0.00 | 3.46 |
Table 5. Precision Analysis of Percentages of Abnormal Signal Patterns for Vysis LSI ATM SpectrumGreen Probe [del(11q22.3)(1 signal)]ª
" The mean and standard deviations are represented as percent abnormal signal patterns.
b Total variance is the sum of the other variance components.
° Positive specimen near the normal cut-off.
、 ·
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| Study | Sample | Category | n | Mean | Between-Day(Within Lot)Component SD | Between-LotComponentSD | Total SDb |
|---|---|---|---|---|---|---|---|
| 1 | 1 | Negative | 7 | 3.5 | 1.12 | 1.10 | 1.57 |
| 1 | 2 | Negative | 7 | 2.2 | 1.69 | 0.00 | 1.69 |
| 1 | 3 | Positive | 7 | 66.1 | 6.13 | 0.00 | 6.13 |
| 1 | 4 | Positive | 7 | 20.1 | 4.88 | 0.00 | 4.88 |
| 1 | 5 | Negative | 7 | 0.6 | 0.83 | 0.00 | 0.83 |
| 1 | 6 | Negative | 7 | 0.4 | 0.64 | 0.00 | 0.64 |
| 1 | 7 | Positive | 7 | 84.9 | 4.14 | 0.00 | 4.14 |
| 1 | 8 | Negative | 7 | 0.6 | 0.97 | 0.00 | 0.97 |
| 1 | 9 | Negative | 7 | 2.3 | 2.08 | 0.00 | 2.08 |
| 1 | 10 | Positive | 7 | 53.6 | 8.57 | 0.00 | 8.57 |
| 2 | 5 | Positivec | 9 | 5.8 | 2.49 | 0.00 | 2.49 |
| 2 | 6 | Positivec | 9 | 14.9 | 2.46 | 1.15 | 2.72 |
Table 6. Precision Analysis of Percentages of Abnormal Signal Patterns for Vysis LSI D13S319 SpectrumOrange Probe [del(13q14.3) (1 signal)]ª
ª The mean and standard deviations are represented as percent abnormal signal patterns.
b Total variance is the sum of the other variance components.
" Positive specimen near the normal cut-off.
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
510(k) Summary Page 11 of 20
.
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| Between-Day(Within Lot)Component SD | Between-LotComponentSD | ||||||
|---|---|---|---|---|---|---|---|
| Study | Sample | Category | n | Mean | Total SDb | ||
| 1 | 1 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 2 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 3 | Negative | 7 | 0.2 | 0.21 | 0.38 | 0.44 |
| 1 | 4 | Negative | 7 | 0.1 | 0.40 | 0.00 | 0.40 |
| 1 | 5 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 6 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 7 | Negative | 7 | 0.2 | 0.32 | 0.00 | 0.32 |
| 1 | 8 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 9 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 10 | Positive | 7 | 18.4 | 6.32 | 0.87 | 6.38 |
| 2 | 8 | Positivec | 9 | 19.8 | 2.53 | 0.00 | 2.53 |
Table 7. Precision Analysis of Percentages of Abnormal Signal Patterns for Vysis LSI D13S319 SpectrumOrange Probe [del(13q14.3) (0 signals)]ª
- The mean and standard deviations are represented as percent abnormal signal patterns.
b Total variance is the sum of the other variance components.
· Positive specimen near the normal cut-off.
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| Study | Sample | Category | n | Mean | Between-Day(Within Lot)Component SD | Between-LotComponentSD | Total SDb |
|---|---|---|---|---|---|---|---|
| 1 | 1 | Negative | 7 | 0.2 | 0.42 | 0.00 | 0.42 |
| 1 | 2 | Negative | 7 | 0.2 | 0.60 | 0.00 | 0.60 |
| 1 | 3 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 4 | Negative | 7 | 0.1 | 0.40 | 0.00 | 0.40 |
| 1 | 5 | Positive | 7 | 21.6 | 6.07 | 0.00 | 6.07 |
| 1 | 6 | Positive | 7 | 70.7 | 4.06 | 1.07 | 4.19 |
| 1 | 7 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 8 | Positive | 7 | 67.3 | 4.84 | 0.00 | 4.84 |
| 1 | .9 | Negative | 7 | 0.0 | 0.00 | 0.00 | 0.00 |
| 1 | 10 | Negative | 7 | 0.1 | 0.20 | 0.00 | 0.20 |
| 2 | 3 | Positivec | 9 | 8.3 | 2.77 | 0.00 | 2.77 |
| 2 | 4 | Positivec | 9 | 3.6 | 3.05 | 0.00 | 3.05 |
Table 8. Precision Analysis of Percentages of Abnormal Signal Patterns for Vysis CEP 12 SpectrumGreen Probe {trisomy 12 (3 signals)]4
" The mean and standard deviations are represented as percent abnormal signal patterns.
6 Total variance is the sum of the other variance components.
- Positive specimen near the normal cut-off.
Reproducibility
Three individual laboratories tested a blinded 20-member slide panel consisting of specimens representing each of the five Dohner classifications:
- . 13q- (monosomy or nullisomy) as sole abnormality
- No cytogenetic abnormality .
- . +12 without 17p- or 11q-
- . 11q- without 17p-
- . 17p-
The analysis was conducted with 10 slides per day for two days for a total of six runs across three testing sites. The same lots of Vysis LSI TP53 SpectrumOrange/ATM SpectrumGreen and LSI D13S319 SpectrumOrange/13q34 SpectrumAqua/CEP 12
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
510(k) Summary Page 13 of 20
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SpectrumGreen Probes were tested at each laboratory. Results shown in Table 9 show an overall agreement with specimens representing each of the five Dohner classifications.
| Disagreeᵃ | Overall | ||||||
|---|---|---|---|---|---|---|---|
| Probe (Abnormality) | Site 1 | Site 2 | Site 3 | NumberAgreeᵃ | NumberTested | PercentAgreement | |
| TP53 (17p-) | 0 | 0 | 0 | 20 | 20 | 100 | |
| ATM (11q-) | 2 | 0 | 0 | 18 | 20 | 90 | |
| CEP 12 (+12) | 0 | 0 | 0 | 20 | 20 | 100 | |
| D13S319 1x (13q-) | 2 | 0 | 0 | 18 | 20 | 90 | |
| D13S319 2x (13q-) | 0 | 1 | 1 | 18 | 20 | 90 |
Table 9. Overall Agreement, Site to Site by Probe
ª Disagree = Number of specimens for which one's site result did not agree with the other sites' results.
·
.
Agree = Number of specimens for which all 3 sites agreed on results.
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The reproducibility of the Vysis CLL FISH Probe Kit for each Prognostic Category demonstrated almost perfect agreement'4 using a generalized kappa statistic among the three testing sites (Table 10).
| Prognostic Category | ||
|---|---|---|
| Sample | Intermediate/Good Prognosisa | Poor Prognosisa |
| 1 | 3 | 0 |
| 2 | 3 | 0 |
| 3 | 3 | 0 |
| 4 | 3 | 0 |
| 5 | 3 | 0 |
| 6 | 3 | 0 |
| 7 | 3 | 0 |
| 8 | 3 | 0 |
| 9 | 3 | 0 |
| 10 | 3 | 0 |
| 11 | 3 | 0 |
| 12 | 3 | 0 |
| 13 | 0 | 3 |
| 14 | 1 | 2 |
| 15 | 0 | 3 |
| 16 | 1 | 2 |
| 17 | 0 | 3 |
| 18 | 0 | 3 |
| 19 | 0 | 3 |
| 20 | 0 | 3 |
| Kappa | 0.86 | |
| Strength | Almost Perfect14 |
Table 10. Vysis CLL FISH Probe Kit Reproducibility Based on Generalized Kappa Statistic
ª Number of sites based on Prognostic Category.
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Additionally, the data were analyzed by site and probe using the Fisher's Exact Test (Tables 11 and 12).
| Site | Prognostic Category | Intermediate/Gooda | Poora |
|---|---|---|---|
| 1 | 12 | 8 | |
| 2 | 12 | 8 | |
| 3 | 14 | 6 | |
| p-value | 0.8396 |
Table 11. Vysis CLL FISH Probe Kit Reproducibility by Site Based on Fisher's Exact Test
ª Number of panel members based on Prognostic Category.
| Table 12. Vysis CLL FISH Probe Kit Reproducibility by Probe Based on Fisher's | ||||
|---|---|---|---|---|
| Exact Test | ||||
| Probe (Abnormality) | Site | AbnormalityDetected | NoAbnormalityDetected | p -value |
| TP 53 (17p-) | 1 | 4 | 16 | 1.0000 |
| 2 | 4 | 16 | ||
| 3 | 4 | 16 | ||
| ATM (11q-) | 1 | 2 | 18 | 0.7495 |
| 2 | 4 | 16 | ||
| 3 | 4 | 16 | ||
| D13S319 1X (13q- [x1]) | 1 | 6 | 14 | 0.8396 |
| 2 | 8 | 12 | ||
| 3 | 8 | 12 | ||
| D13S319 2x (13q- [x2]) | 1 | 2 | 18 | 1.000 |
| 2 | 3 | 17 | ||
| 3 | 3 | 17 | ||
| CEP 12 (+12) | 1 | 5 | 15 | 1.000 |
| 2 | 5 | 15 | ||
| 3 | 5 | 15 |
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
510(k) Summary Page 16 of 20
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Clinical Utility
The traditional Rai and Binet clinical CLL staging systems are based on disease burden and have been useful for assigning patients to groups having similar survival times. 1911 These systems, however, are not effective in predicting survival in early-stage disease when most CLL cases are diagnosed. This has led to development of newer molecular markers in an attempt to differentiate those patients who are prone to rapid progression from those who have indolent disease.
In a pivotal study conducted by Döhner et al, titled "Genomic Aberrations and Survival in Chronic Lymphocytic Leukemia," genomic alterations as determined by FISH were found to be predictive for disease progression and overall survival. Multiple studies support the conclusion of Dohner et al that loss of 11q markers predicts reduced survival times as compared to other Dohner groups as determined by FISH aberrations.68 Such studies have led to the inclusion of FISH testing in the National Comprehensive Cancer Network (NCCN) practice guidelines as a means to determine CLL prognosis. 12
In a 2006 prospective study of 151 patients by Shanafelt, et al. utilizing Vysis FISH probes, a correlation was established between overall survival and FISH risk category for CLL at diagnosis. Patients were divided into two prognostic groups. They were assigned to the good/intermediate FISH prognosis group if there were no chromosomal aberrations or if only 13q- and/or +12 aberrations were present. If a chromosomal aberration of 17por I I q- was present, the patient was placed in the poor FISH prognosis group. Poor vs. good/intermediate FISH (P=0.004), age at diagnosis (P=0.0006), and Rai stage (P=0.0026) were each significantly associated with overall survival from diagnosis in univariate analysis. When all factors were included in multivariable Cox regression model, each of three factors still remained significant: poor vs. good/intermediate FISH (P=0.00022), age at diagnosis (P=0.000024); and Rai stage (P=0.00012).
The clinical utility of the Vysis CLL FISH Probe Kit has been established primarily from its high concordance with the assay employed in the publication of Shanafelt et al.
510(k) Summary Page 17 of 20
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Also, as noted in the Shanafelt study, all patients with the 17p- abnormality had between 24-94% of cells with this abnormality. Therefore, the effect of 17p- at very low levels could not be determined. In a recent publication on untreated 17p- CLL patients, Tam et al reported a 3-year overall survival of 92% for patients with < 25% 17p-deleted nuclei, vs. 54% for patients with ≥ 25% 17p-deleted nuclei (P=0.007).12
The NCCN CLL practice guideline (v.2.2011) states that prognostic CLL FISH abnormalities noted on page CLLS-A of that document (including abnormalities tested for by this kit) may not have the noted clinical prognostic impact when present in less than 10% of cells. 13
Method Concordance
This study establishes the clinical validity of the AMT by demonstrating concordance to the Reference FISH Test (RFT) used in the Shanafelt study.8 The clinical validity of the RFT is documented via peer-reviewed literature.
This study analyzed 64 specimens whose Döhner classifications were based on previous results using the RFT. Table 13 shows the distribution of specimens by Döhner classification.
| Döhner Classification | Number of Specimens Analyzed |
|---|---|
| 13q- (monosomy or nullisomy) as soleabnormality | 13 |
| No cytogenetic abnormality | 12 |
| +12 without 11q- or 17p- | 14 |
| 11q- without 17p- | 18 |
| 17p- | 7 |
| Table 13. Dohner Classification of Specimens Analyzed in Concordance Study | ||||||
|---|---|---|---|---|---|---|
| ---------------------------------------------------------------------------- | -- | -- | -- | -- | -- | -- |
The specimens were de-identified and randomized before testing was conducted with the AMT and the RFT.
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
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A Dohner classification was assigned to each AMT and RFT result, and the Prognostic Category was determined.
The percent agreement between the AMT and RFT for Prognostic Category was 97% (62/64) with a lower bound of the one-sided 95% confidence interval of 90% (Table 14). Overall agreement was defined as the percentage of specimens that had the same Prognostic Category when testing using the RFT and the AMT.
| Reference FISH Test (RFT) | |||
|---|---|---|---|
| Abbott Molecular Test (AMT) | Intermediate/Good | Poor | Total |
| Intermediate/Good | 38 | 1 | 39 |
| Poor | 1 | 24 | 25 |
| Total | 39 | 25 | 64 |
Table 14. Concordance between AMT and RFT for Prognostic Category"
a The values in this table represent the number of specimens.
References
-
- Shanafelt TD, Byrd JC, Call TG, et al. Narrative review: initial management of newly diagnosed, early-stage chronic lymphocytic leukemia. Ann Intern Med. 2006;145:435-7.
-
- Méhes G. Chromosome abnormalities with prognostic impact in B-cell chronic lymphocytic leukemia. Pathol Oncol Res. 2005;11(4):205-10.
-
- Stilgenbauer S. Bullinger L. Lichter P. et al. Genetics of chronic lymphocytic leukemia: genomic aberrations and V(H) gene mutation status in pathogenesis and clinical course. Leukemia. 2002;16(6):993-1007.
-
- Ripollés L, Ortega M, Ortuño F, et al. Genetic abnormalities and clinical outcome in chronic lymphocytic leukemia. Cancer Genet Cytogenet. 2006;171(1):57-64.
-
- Dohner H, Stilgenbauer S, Benner A, et al. Genomic aberrations and survival in chronic lymphocytic leukemia. N Eng J Med. 2000:343(26):1910-6.
-
- Kröber A, Seiler T, Benner A, et al. V(H) mutation status, CD38 expression level, genomic aberrations, and survival in chronic lymphocytic leukemia. Blood. 2002; 100(4):1410-6.
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-
- Oscier DG. Gardiner AC, Mould SJ, et at. Multivariate analysis of prognostic factors in CLL: clinical stage, IGVH gene mutational status, and loss or mutation of the p53 gene are independent prognostic factors. Blood. 2002;100(4):1177-84.
-
- Shanafelt TD, Witzig TE, Fink SR, et al. Prospective evaluation of clonal evolution during long-term follow-up of patients with untreated early-stage chronic lymphocytic leukemia. J Clin Oncol. 2006;24(28):4634-41.
-
- Wiktor AE. Van Dyke DL, Stupca PJ, et al. Preclinical validation of fluorescence in situ hybridization assays for clinical practice. Genet Med. 2006;8(1):16-23.
-
- Rai KR. Sawitsky A, Cronkite EP, et al. Clinical staging of chronic lymphocytic leukemia. Blood. 1975;46(2):219-34.
-
- Binet JL, Auguier A, Dighiero G, et al. A new prognostic classification of chronic lymphocytic leukemia derived from a multivariate survival analysis. Cancer. 1981; 48(1):198-206.
-
- Tam CS, Shanafelt TD, Wierda WG, et al. De novo deletion 17p13.1 chronic lymphocytic leukemia shows significant clinical heterogeneity: the MD Anderson and Mayo Clinic experience. Blood. 2009;114(5):957-64
-
- NCCN Clinical Practice Guidelines in Oncology™ NON-HODGKIN'S LYMPHOMAS (Version 2.2011). ©2011 National Comprehensive Cancer Network, Inc. Available at: NCCN.org. Accessed [December 20, 2009].
-
- Landis JR, Koch GG. The measurement of observer agreement for categorical data. Biometrics. 1977;33:159-174.
Vysis CLL FISH Probe Kit 510(k) K100015 August 9, 2011
· 510(k) Summary Page 20 of 20
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Image /page/20/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of two main elements: a circular text element and an abstract symbol. The text element is arranged in a circle and reads "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA". To the right of the text is an abstract symbol that resembles a stylized bird or a flame.
Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993
Abbott Molecular Inc. c/o Ms. Nancy Bengtson Manager, Global Regulatory Affairs 1300 E. Touhy Avenue Des Plaines, IL, 60018
AUG 0 9 2011
Re: K100015
Trade/Device Name: Regulation Number: Regulation Name: Regulatorv Class: Product Code: Dated: Received:
Vysis CLL FISH Probe Kit 21 CFR §866.6040 Gene expression profiling test system for breast cancer prognosis Class II OVQ July 21, 2011 July 22, 2011
Dear Ms. Bengtson:
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 class II (Special Controls), 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); medical device reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). 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
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Page 2 - Ms. Nancy Bengtson
predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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 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/cdrh/industry/support/index.html.
Sincerely yours,
Marci M. Chan
Maria M. Chan, Ph.D Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
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Indications for Use Statement
510(k) Number:
Device Name: Vysis CLL FISH Probe Kit
Indications for Use:
The Vysis CLL FISH Probe Kit is intended to detect deletion of the LSI TP53, LSI ATM, and LSI D13S319 probe targets and gain of the D12Z3 sequence in peripheral blood specimens from untreated patients with B-cell chronic lymphocytic leukemia (CLL). The assay may be used to dichotomize CLL (the 13q-, +12, or normal genotype group versus the 11g- or 17p- group) and may be used as an aid in determining disease prognosis in combination with additional biomarkers, morphology and other clinical information. The Vysis CLL FISH Probe Kit is not intended for use in selection of therapy or in monitoring of residual disease.
Prescription Use X (Part 21 CFR 801 Subpart D)
AND/OR
Over-The-Counter Use (21 CFR 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
510(k) K100015
Page 1 of
§ 866.6040 Gene expression profiling test system for breast cancer prognosis.
(a)
Identification. A gene expression profiling test system for breast cancer prognosis is a device that measures the ribonucleic acid (RNA) expression level of multiple genes and combines this information to yield a signature (pattern or classifier or index) to aid in prognosis of previously diagnosed breast cancer.(b)
Classification. Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Gene Expression Profiling Test System for Breast Cancer Prognosis.” See § 866.1(e) for the availability of this guidance document.