The Vysis EGR1 FISH Probe Kit is intended to detect deletion of the LSI EGR1 probe target on chromosome 5q in bone marrow specimens and to be used, in addition to cytogenetics, other biomarkers, morphology and other clinical information, at the time of acute myeloid leukemia (AML) diagnosis as an aid in determining prognosis. Deletion of chromosome 5q has been associated with an unfavorable prognosis in AML patients.

Device Description

The Vysis EGR1 FISH Probe Kit uses fluorescence in situ hybridization (FISH) DNA probe technology to determine deletion status of the LSI EGR1 (containing early growth response 1 gene; location chromosome 5q31) probe target in AML specimens. The Vysis EGR1 FISH Probe Kit also contains the LSI D5S23, D5S721 probe (location chromosome 5p15.2) and serves as a control. The kit consists of one vial containing two DNA FISH probes and four general purpose reagents sufficient to process 20 specimens.

AI/ML Overview

Here's a summary of the acceptance criteria and the study details for the Vysis EGR1 FISH Probe Kit, based on the provided text:

Vysis EGR1 FISH Probe Kit: Acceptance Criteria and Study Details

1. Table of Acceptance Criteria and Reported Device Performance

Acceptance Criteria	Reported Device Performance
Analytical Specificity:
LSI D5S23, D5S721 Probe	100% (95% CI: 98, 100)
EGR1 Probe	100% (95% CI: 98, 100)
Analytical Sensitivity:
LSI EGR1/D5S23, D5S721	99.6% (95% CI: 99.4, 99.7) for 2R2G signal pattern
Normal Cut-off:
Max. 1R2G patterns	Specimens exceeding 6% or 12 1R2G patterns per 200 nuclei considered abnormal. No normal specimens produced 1R2G signals at or above 6%.
Reproducibility (Site-to-Site):
Overall Agreement (High Positive)	100%
Overall Agreement (Low Positive)	100%
Overall Agreement (Normal)	98% (one discordant result out of 60 observations)
Reproducibility (Lot-to-Lot):
Overall Agreement (High Positive)	100%
Overall Agreement (Low Positive)	100%
Overall Agreement (Normal)	100%
Clinical Utility (Agreement with Karyotype):
Overall Agreement	98.90% (179/181) (95% CI: 96.06%-99.70%)
Negative Percent Agreement	100% (171/171) (95% CI: 97.80%-100.00%)
Positive Percent Agreement	80% (8/10) (95% CI: 49.02%-94.33%)

2. Sample Size Used for the Test Set and Data Provenance

Analytical Specificity: Metaphase chromosomes from 5 karyotypically normal males.
Analytical Sensitivity: Interphase nuclei from 25 bone marrow specimens (karyotypically normal or 5p15 and 5q31 deletion-free).
Verification of Normal Cut-off: Interphase nuclei from 25 bone marrow specimens (karyotypically normal or 5p15.2 and 5q31 deletion-free).
Reproducibility (Site-to-Site): 2 high-positive, 2 low-positive, and 2 normal specimens (bone marrow cells mixed to create positive controls).
Reproducibility (Lot-to-Lot): Same 2 high-positive, 2 low-positive, and 2 normal specimens.
Clinical Utility (Agreement with Karyotype): 181 bone marrow specimens from an Eastern Cooperative Oncology Group (ECOG) clinical trial (E1900) for AML diagnosis. This appears to be retrospective data from a specific clinical trial. The country of origin is implied to be the US given the ECOG trial.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications

Analytical Specificity: "one technologist" evaluated 100 metaphase nuclei per sample. No specific qualifications are provided for this technologist beyond being a "technologist."
Analytical Sensitivity: "two technologists" evaluated 100 nuclei per specimen. No specific qualifications provided.
Verification of Normal Cut-off: "two technologists" evaluated 100 nuclei per specimen. No specific qualifications provided.
Clinical Utility (Agreement with Karyotype): The ground truth for this section is based on "karyotype" results. While the text refers to a publication by Vance et al. from an ECOG clinical trial, it does not explicitly state the number or qualifications of the experts establishing the karyotype ground truth for this specific study. However, karyotyping is a specialized cytogenetic analysis performed by trained professionals (cytogeneticists/pathologists).

4. Adjudication Method for the Test Set

Analytical Specificity, Sensitivity, Normal Cut-off: The text mentions evaluation by one or two technologists. For cases where two technologists evaluated, it doesn't explicitly state an adjudication method if discrepancies occurred. It simply states "Each technologist evaluated..." or "evaluated by two technologists." This suggests independent evaluation, but no formal adjudication process is described for reconciling differences.
Reproducibility: A mean and standard deviation were calculated across replicates and sites/lots, indicating a statistical comparison of results, not a consensus-based adjudication of individual cases.
Clinical Utility (Agreement with Karyotype): The comparison is against established karyotype results, which are considered the ground truth. There's no further adjudication stated for these 181 specimens beyond that initial karyotype.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No Multi-Reader Multi-Case (MRMC) comparative effectiveness study was done comparing human readers with AI assistance vs. without AI assistance. This device is a FISH probe kit, not an AI-assisted diagnostic tool.

6. Standalone (Algorithm Only) Performance

This device is a physical FISH probe kit designed for laboratory use with microscopic interpretation by trained personnel. It does not involve a standalone algorithm or AI for interpretation. Performance metrics like analytical specificity, sensitivity, and reproducibility are for the kit's ability to produce expected signal patterns when interpreted by human technologists.

7. Type of Ground Truth Used

Analytical Specificity: Metaphase chromosomes from karyotypically normal individuals, with "correct locus" hybridization serving as the ground truth.
Analytical Sensitivity: Expected normal signal pattern (2R2G) in interphase nuclei from karyotypically normal or deletion-free bone marrow specimens.
Verification of Normal Cut-off: Normal bone marrow specimens that were karyotypically normal or 5p15.2 and 5q31 deletion-free.
Reproducibility: Known status of specimens (high-positive, low-positive, normal) created by mixing positive and normal bone marrow cells.
Clinical Utility (Agreement with Karyotype): Cytogenetic results (karyotype) from clinical AML diagnosis ("-5/del5q" vs. normal karyotype).

8. Sample Size for the Training Set

The document does not explicitly describe a separate "training set" in the context of machine learning or algorithm development, as this device is a probe kit. The studies described are for analytical validation and clinical correlation.

9. How the Ground Truth for the Training Set Was Established

Not applicable, as there's no mention of a traditional "training set" for an algorithm in this submission. The validation studies rely on established biological principles and comparisons to conventional cytogenetic methods.

Summary

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510(k) Summary

AUG 2 9 2011

510(k) Number K091960

Device Name: Vysis EGR1 FISH Probe Kit (Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes) (List No. 4N37-020)

Purpose of the Submission

The purpose of this 510(k) is to gain clearance to market the Vysis EGR1 FISH Probe Kit (List No. 4N37-020).

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, 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 25, 2011

Manufacturer

Abbott Molecular Inc. is the legal manufacturer of the Vysis EGR 1 FISH Probe Kit (List No. 4N37-020).

Name:	Timothy Zurow, PhD
Title:	Director of Manufacturing Operations
Telephone .	(224) 361-7379
Fax:	(224) 361-7038
Email:	timothy.zurow@abbott.com

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Abbott Molecular Inc. Address: 1300 E. Touhy Avenue Des Plaines, IL 60018

Establishment Registration No.: 3005248192

Intended Use

The Vysis EGR1 FISH Probe Kit is intended to detect deletion of LSI EGR1 probe target on chromosome 5q in bone marrow specimens and to be used, in addition to cytogenetics, other biomarkers, morphology and other clinical information, at the time of acute myeloid leukemia (AML) diagnosis as an aid in determining prognosis. Deletion of chromosome 5q has been associated with an unfavorable prognosis in AML patients.

Trade Name

Vysis EGR1 FISH Probe Kit (Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes) (List No. 4N37-020)

Common Name

Vysis EGR1 Fluorescence In Situ Hybridization (FISH) Probe Kit

Classification

Class II

Regulation Number

21 CFR8866.6040, Gene expression profiling test system for breast cancer prognosis

Product Code

OWK, Early growth response 1 (EGR1) kit

Predicate Device(s)

Immunology

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Comparison with Predicate Device(s)

Similarities
Item	Device	Predicate (K100015)
Intended Use	Determination of deletion status as an aid in determining prognosis	Same
Technology	FISH	Same
Differences
Item	Device	Predicate (K100015)
Patient Population	Acute myeloid leukemia (AML) patients	Chronic lymphocytic leukemia (CLL) patients
Probe Targets	LSI EGR1	LSI TP53LSI ATMLSI D13S319CEP 12

Similarities

Device Description

DNA Probe Description

Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes:

The SpectrumOrange-labeled LSI EGR1 probe, approximately 209 kb in length (chr5:137682107-137890637; March 2006 Assembly; UCSC Human Genome Browser'), is located at 5q31 and contains the complete EGR1 gene.

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The SpectrumGreen-labeled LSI D5S23, D5S721 probe, approximately 561 kb in length (chr5:9450109-10011407; March 2006 Assembly; UCSC Human Genome Browser'), is located at 5p15.2.

The Vysis EGR1 FISH Probe Kit (List No. 4N37-020) consists of one vial containing two DNA FISH probes and four general purpose reagents sufficient to process 20 specimens.

. Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes
. Vysis LSI/WCP Hybridization Buffer
. DAPI II Counterstain
. NP-40
. 20X SSC Salt

Background on Acute Myeloid Leukemia (AML) Disease

Deletion of chromosome 5q as detected by cytogenetics is a recurring abnormality in AML. A commonly deleted segment on chromosome band 5q31 has been identified and the early growth response 1 (EGR1) gene is among the candidate genes in this segment.2 A study suggests that haploinsufficiency of EGR1 may play a role in leukemogenesis.3 The Vysis locus-specific identifier (LSI) EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes, components of the Vysis EGR1 FISH Probe Kit, have been used in several studies to detect EGR1 deletions.47 A study, conducted as part of an Eastern Cooperative Oncology Group (ECOG) clinical trial, demonstrated the utility of the interphase fluorescence in situ hybridization (FISH) technique to stratify patients into cytogenetic risk categories at diagnosis of AML using the Vysis LSI EGR1 SpectrumOrange/D5S23. D5S721 SpectrumGreen Probes in conjunction with several additional FISH probes. In addition, the prognostic importance of chromosome 5 abnormalities has been established in several large clinical studies.80 The prognostic clinical utility of detecting specific chromosomal abnormalities in bone marrow specimens from patients diagnosed with AML is firmly established in both standard medical practice guidelines and in the medical literature. The National Comprehensive Cancer Network (NCCN) Practice Guidelines™ for Acute Myeloid Leukemia, which are

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the consensus recommendations of leading US AML experts, states that cytogenetics is the single most important prognostic factor in AML. 4 More specifically, the guidelines provide different risk categories for patients depending upon cytogenetic or molecular abnormalities. As an example, patients with deletion of chromosome 5q are in the 'poor risk' category and may have an unfavorable prognosis.

The Vysis EGR1 FISH Probe Kit uses FISH DNA probe technology to determine deletion status of the probe target for LSI EGR1, and the LSI D5S23, D5S721 probe serves as a control.

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.

Cells attached to microscope slides using standard cytogenetic procedures are used for this assay. The resulting specimen DNA is denatured to single-stranded form and subsequently allowed to hybridize with the LSI EGR1 and LSI D5S23, D5S721 probes. Following hybridization. the unbound probe is removed by a series of washes, and the nuclei are counterstained with DAPI, a DNA-specific stain that fluoresces blue. Hybridization of the LSI EGR1 and LSI D5S23, D5S721 probes is viewed using a fluorescence microscope equipped with appropriate excitation and emission filters. allowing visualization of the orange and green fluorescent signals. In a cell with normal copy numbers of the LSI EGR1 and LSI D5S23, D5S721 probe targets, two SpectrumOrange signals (LSI EGR1) and two SpectrumGreen signals (LSI D5S23, DSS721) will be expected.

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In a cell with the 5q deletion, one SpectrumOrange signal (LSI EGR1) and two SpectrumGreen signals (LSI D5S23, D5S721) will be expected. Enumeration of the orange LSI EGR1 and green LSI D5S23, D5S721 signals provide a mechanism for determining absolute copy number of the probe targets and the presence of the aberrations of interest.

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. The analytical specificity of the Vysis LSI EGR1 D5S23, D5S721 probes for their respective chromosome target loci was established using metaphase chromosomes prepared from peripheral blood cultures of five karyotypically normal males that were pooled prior to dropping on microscope slides. The hybridization location of each FISH signal on chromosomes of 100 consecutive metaphase nuclei 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 analytical 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 of the Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes was 100%, as shown in Table 1.

510(k) Summary Page 6 of 12

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Table 1. Analytical Specificity
	Number of MetaphaseChromosome Signals
Probe	Hybridized tothe CorrectTarget Locus	TotalHybridizedSignals	AnalyticalSpecificity (%)	95%ConfidenceInterval (%)
D5S23, D5S721	200	200	100	(98, 100)
EGR1	200	200	100	(98, 100)

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 the probes in the Vysis EGR1 FISH Probe Kit is 2R2G per nucleus.

The analytical sensitivity of the Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probes was established using interphase nuclei prepared from 25 bone marrow specimens that were either karyotypically normal or 5p15 and 5q31 deletion-free. The orange and green signal patterns of nuclei for 25 specimens were evaluated by two technologists. Each technologist evaluated 100 nuclei per specimen for a total of 200 nuclei per specimen and 5000 scoreable nuclei from normal specimens.

The analytical sensitivity was calculated as the percentage of scoreable interphase nuclei with the expected 2R2G signal pattern at the 95% confidence interval.

The Vysis EGR1 FISH Probe Kit has an analytical sensitivity of 99.6%, as shown in Table 2.

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Table 2. Analytical Sensitivity
	Number of InterphaseChromosome Signals		Analytical Sensitivity (%)
Probe Kit	With ExpectedSignal Pattern	ScoreableSignals	PointEstimate	95% ConfidenceInterval
LSI EGR1/D5S23, D5S721	4979	5000	99.6	(99.4, 99.7)

Verification of Normal Cut-off

The normal cut-off value is defined as the maximum quantity of scoreable interphase nuclei with a specific abnormal signal pattern at which a specimen is considered normal for that signal pattern. The normal cut-off value is expressed in terms of a percentage or the actual number of a specific abnormal FISH signal pattern per the standard number of nuclei tested.

The normal cut-off value for this assay is 6% or 12 1R2G patterns per 200 scoreable interphase nuclei. Specimens exceeding 12 1R2G patterns per 200 scoreable nuclei are considered abnormal for deletion of the Vysis LSI EGR1 probe target. This 6% normal cut-off value was adopted from the publication of Vance et al, who utilized the Vysis LSI ERG1/D5S23, D5S721 probe set in a study that established a high level of agreement between cytogenetics and FISH in 237 blood and bone marrow specimens studied at AML diagnosis.4

In order to confirm that the 6% normal cut-off served well to prevent normal specimens from being called abnormal, the assay was performed on interphase nuclei from 25 bone marrow specimens from either karyotypically normal specimens or 5p15.2 and 5q31 deletion-free specimens. The signal patterns of 200 nuclei were evaluated by counting the number of orange and green signals. Each of two technologists evaluated 100 nuclei per specimen. Among the 25 normal specimens, none produced 1R2G signals at or above the 6% normal cut-off.

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Reproducibility

Two replicates of the assay were run on 2 high-positive, 2 low-positive, and 2 normal specimens at three sites on 5 different days. The positive specimens for the site-to-site study were obtained by mixing positive bone marrow cells with normal bone marrow cells to obtain the desired levels of abnormality. The mean and the standard deviation of the percentage of cells with the 1R2G signal pattern was calculated. Results shown in Table 3 show the overall agreement with the normal/abnormal status of the test specimens. All sites obtained 100% agreement with the known status of all 6 specimens on all 5 days, except one site which had one discordant result for a normal specimen.

Table 3. Overall Agreement, Site to Site
	Number
Category	Agree	Disagree	Total	Percent Agreement
High Positive	60	0	60	100
Low Positive	60	0	. 60	100
Normal	રેતે રહ્યું રહ્યું હતું. સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત કર્યું સ્વિત		60	98

The analysis of variance components for the site-to-site study is shown in Table 4.

Table 4. Site-to-Site Analysis of Variance Components
			Within	Between	Between	Total
Sample	N	Meana	Day SDb	Day SD	Site SD	SD
High Positive 1	30	70.0	3.28	4.01	5.44	7.51
High Positive 2	30	47.6	5.56	0.00	0.74	5.61
Low Positive 1	30	18.1	3.00	3.82	1.03	4.97
Low Positive 2	30	14.9	3.25	1.54	0.00	3.59
Normal 1	30	0.7	0.71	0.00	0.68	0.99
Normal 2	30	0.9	0.66	1.42	0.22	1.59

4 Percentage of cells with 1R2G signal pattern

b SD = Standard deviation

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Using the same specimens from the site-to-site study, four replicates of the assay were run on 2 high-positive, 2 low-positive, and 2 normal specimens using 3 different lots of probe at a single site. The overall agreement with the known normal/abnormal status of the test specimen is shown in Table 5. All replicates using the three probe lots for each of the 6 specimens produced agreement with the known status of the specimens.

Table 5. Overall Agreement, Lot to Lot
	Number			Percent
Category	Agree	Disagree	Total	Agreement
High Positive	24	0	24	100
Low Positive	24	0	24	100
Normal	24	0	24	100

The analysis of variance components for the lot-to-lot study is shown in Table 6.

Table 6. Lot-to-Lot Analysis of Variance Components
Sample	N	Meana	Within Lot SDb	Between Lot SD	Total SD
High Positive 1	12	66.2	7.19	0.00	7.19
High Positive 2	12	47.4	3.69	3.04	4.78
Low Positive 1	12	12.7	4.29	0.00	4.29
Low Positive 2	12	12.3	1.84	1.12	2.15
Normal 1	12	0.0	0.00	0.00	0.00
Normal 2	12	0.1	0.14	0.20	0.25

ª Percentage of cells with 1R2G signal pattern

b SD = Standard deviation

In these reproducibility studies, 84 assays were run on low positive specimens. None of the 84 low-positive assays resulted in a 1R2G signal pattern at or below 6%.

Clinical Utility

The clinical utility of the cytogenetic detection of the deletion of chromosome 5q is correlated with reduced 5-year overall survival in studies by Byrd et al ° and Grimwade et

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al . The Byrd publication demonstrated the prognostic value in 86 patients with a -5/5qabnormality by a median overall survival (OS) of 0.3 years compared to the median OS of 1.3 years associated with patients exhibiting a normal karyotype. Patients with a -5/5qabnormality had a 5-year OS of 6% compared to a 5-year OS of 24% associated with patients exhibiting a normal karyotype. Patients with a -5/5q- abnormality also had a significantly lower complete remission (CR) rate of 31% than the normal karyotype, which resulted in a CR of 68% with a p<0.001. For del(5q), 42 patients had a 5-year OS of 5%, 95% CI (2-13%) and median overall survival of 0.3 years.

Grimwade et al, showed prognostic value in 28 patients for 5q-. Cytogenetic abnormalities in the Medical Research Council (MRC) AML 10 clinical trial for del(5q) patients showed 5-year OS of 11% and CR of 57% in the adverse risk group. These values varied significantly from the no abnormality, or normal karyotype group, which had 5-year OS of 42% and CR of 88% (p<0.001).

The publication "Utility of interphase FISH to stratify patients into cytogenetic risk categories at diagnosis of AML in an Eastern Cooperative Oncology Group (ECOG) clinical trial (E1900)'' by Vance et al', establishes linkage between cytogenetic results and the Vysis EGR1 FISH Probe kit. When 181 bone marrow specimens were compared to cytogenetic results at the >6% cut-off, there was overall agreement of 98.90% (179/181) (95% CI-96.06%-99.70%), negative percent agreement of 100% (171/171) (95% CI-97.80%-100.00%) and positive percent agreement of 80% (8/10) (95% CI-49.02%-94.33%). Results are presented below:

		Karyotype -5/del5q
		Positive	Negative	Total
FISH (1R2G- 5q deletion signalpattern)	Positive	8	0	8
	Negative	2a,b	171	173
	Total	10	171	181

a Cytogenetic result was -5/del(5q). FISH signal pattern was 44% 1R1G (monosomy of chromosome 5).

6 Cytogenetic result was -5/del(5q). FISH signal pattern was 1% 1R2G. False negative results.

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BIBLIOGRAPHY

1. Genome Bioinformatics Group of UC Santa Cruz. The UCSC Genome Browser. © The Regents of the University of California. Available at: http://genome.ucsc.edu/cgibin/hgGateway?hgsid=185806115&clade=mammal&org=Human&db=hg18. Accessed [February 9, 2011].
1. Lai F. Godley LA, Joslin J, et al. Transcript map and comparative analysis of the 1.5-Mb commonly deleted segment of human 5q31 in malignant myeloid diseases with a del(5q). Genomics. 2001;71(2):235-45.
1. Joslin JM, Fernald AA, Tennant TR, et al. Haploinsufficiency of EGR1, a candidate gene in the del(5q), leads to the development of myeloid disorders. Blood. 2007;110(2):719-26.
1. Vance GH, Kim H, Hicks GA, et al. Utility of interphase FISH to stratify patients into cytogenetic risk categories at diagnosis of AML in an Eastern Cooperative Oncology Group (ECOG) clinical trial (E1900). Leuk Res. 2007;31:605-9.
1. Herry A, Douet-Guilbert N, Morel F, et al. Redefining monosomy 5 by molecular cytogenetics in 23 patients with MDS/AML. Eur J Haematol. 2007;78:457-67.
1. Bram S, Swolin B, Rodjer S, et al. Is monosomy 5 an uncommon aberration? Fluorescence in situ hybridization reveals translocations and deletions in myelodysplastic syndromes or acute myelocytic leukemia. Cancer Genet Cytogenet. 2003;142:107-14.
1. Bram S. Rodjer S. Swolin B. Several chromosomes involved in translocations with chromosome 5 shown with fluorescence in situ hybridization in patients with malignant myeloid disorders. Cancer Genet Cytogenet. 2004:155:74-8. 1
1. Byrd JC, Mrozek K, Dodge RK, et al. Pretreatment cytogenetic abonormalities are predictive of induction success, cumulative incidence of relapse, and overall survival in adult patients with de novo acute myeloid leukemia: results from Cancer and Leukemia Group B (CALGB 8461). Blood. 2002;100:4325-36.
1. Grimwade D, Walker H, Oliver F, et al. The importance of diagnostic cytogenetics on outcome in AML: analysis of 1,612 patients entered into the MRC AML 10 trial. The Medical Research Council Adult and Children's Leukaemia Working Parties. Blood. 1998;92(7):2322-33.
1. NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines™). Acute Myeloid Leukemia. (Version 2.2011). ©2010 National Comprehensive Cancer Network, Inc. Available at: NCCN.org. Accessed [February 9, 2011].

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Image /page/12/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" arranged around the perimeter. Inside the circle is a stylized image of an eagle or other bird with outstretched wings, depicted in a flowing, abstract manner.

Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993

Abbott Molecular Inc. c/o Ms. Pamela L. Swatkowski Program Manager Regulatory Affairs 1300 E. Touhy Avenue Des Plaines, IL 60018

AUG 2 9 2011

Re: K091960

Trade/Device Name: Vysis EGR1 FISH Probe Kit Regulation Number: 21 CFR §866.6040 Regulation Name: Gene expression profiling test system for breast cancer prognosis Regulatory Class: Class II Product Code: OWK Dated: February 24, 2011 Received: February 25, 2011

Dear Ms. Swatkowski:

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 (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice

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Page 2 - Ms. Pamela Swatkowski

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 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,

7.V.L. In Chan

Maria Chan, PhD 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 EGR1 FISH Probe Kit (Vysis LSI EGR1 SpectrumOrange/D5S23, D5S721 SpectrumGreen Probe)

Indications for Use:

The Vysis EGR I FISH Probe Kit is intended to detect deletion of the LSI EGR1 probe target on chromosome 5q in bone marrow specimens and to be used, in addition to cytogenetics, other biomarkers, morphology and other clinical information, at the time of acute myeloid leukemia (AML) diagnosis as an aid in determining prognosis. Deletion of chromosome 5q has been associated with an unfavorable prognosis in AML patients.

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) K091960

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Regulation Number and Section

§ 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.