Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device is a FISH probe kit for specimen characterization, and the interpretation is performed by a qualified pathologist or cytogeneticist. There is no mention of automated analysis or algorithms that would suggest AI/ML.

Therapeutic

No
The 'Intended Use / Indications for Use' section explicitly states, "This device is not intended for high-risk uses such as selecting therapy, predicting therapeutic response or disease screening." This indicates it's not a therapeutic device.

Diagnostic

No

The "Intended Use / Indications for Use" section explicitly states, "The use of this device for diagnosis, prognosis, monitoring or risk assessment has not been established." It also says, "This device is not intended for high-risk uses such as selecting therapy, predicting therapeutic response or disease screening," and that it is "intended for specimen characterization."

SaMD (Software as a Medical Device)

No

The device is a FISH probe kit, which is a laboratory reagent and not a software-only medical device. It consists of DNA probes and other chemical reagents.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states that the device is for "specimen characterization" in bone marrow and peripheral blood specimens from patients with acute myeloid leukemia or myelodysplastic syndrome. This involves testing biological specimens outside of the body to obtain information about a patient's health.
Device Description: The device is a "FISH Probe Kit" which contains reagents used to perform a test on biological samples.
Performance Studies: The document describes analytical performance studies (specificity, sensitivity, reproducibility) conducted on biological specimens (bone marrow and peripheral blood).
Predicate Device: The predicate device listed (K123951; Vysis EGR1 FISH Probe Kit - SC) is also a FISH probe kit intended for specimen characterization, which is a common type of IVD.

While the intended use explicitly states it's not for diagnosis, prognosis, monitoring, or risk assessment, the act of characterizing a specimen from a patient using a test performed outside the body falls under the definition of an In Vitro Diagnostic device. The "Specimen Characterization" indication is a specific category within IVDs.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Vysis D7S486/CEP 7 FISH Probe Kit is a device intended for specimen characterization, and detects the LSI D7S486 probe target on chromosome 7q31 and the CEP 7 probe target on chromosome 7p11.1-q11.1 in bone marrow and peripheral blood specimens from patients with acute myeloid leukemia or myelodysplastic syndrome. The assay results are intended to be interpreted by a qualified pathologist or cytogeneticist. This device is not intended for high-risk uses such as selecting therapy, predicting therapeutic response or disease screening. The use of this device for diagnosis, prognosis, monitoring, or risk assessment has not been established.

Product codes

PFG

Device Description

The Vysis D7S486/CEP 7 FISH Probe Kit is for specimen characterization and detects the LSI D7S486 (7q31) probe target on chromosome 7q31 and CEP 7 probe target chromosome 7p11.1-q11.1 in bone marrow and peripheral blood specimens.

DNA Probe Description

Vysis LSI D7S486 SpectrumOrange/ CEP 7 SpectrumGreen Probes:

The SpectrumOrange labeled LSI D7S486 probe is approximately 308 kb in length (chr7:115983468-115675366; February 2009 Assembly UCSC Human Genome Browser).

The SpectrumGreen labeled CEP 7 probe targets the D7Z1 alpha satellite sequence at the centromere of chromosome 7.

The Vysis D7S486/CEP 7 FISH Probe Kit (List No. 04N78-020) consists of a mixture of two DNA FISH probes and four general reagents sufficient to process 20 assays.

. Vysis LSI D7S486 SpectrumOrange/ CEP 7 SpectrumGreen Probes
. Vysis LSI/WCP Hybridization Buffer
. DAPI II Counterstain
NP-40 .
. 20X SSC

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

chromosome 7q31 and chromosome 7p11.1-q11.1

Indicated Patient Age Range

Not Found

Intended User / Care Setting

qualified pathologist or cytogeneticist

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)

Analytical Specificity:
Established using metaphase chromosomes prepared from peripheral blood cultures of 4 male and 1 female karyotypically normal specimen slides. The hybridization location of each FISH signal on chromosomes of 100 consecutive metaphase nuclei was evaluated by 1 technologist for a total of 200 target loci per probe.
The analytical specificity of the Vysis LSI D7S486/CEP 7 FISH Probe Kit was 100%.

Analytical Sensitivity:
Established using interphase nuclei prepared from 25 bone marrow and 25 peripheral blood specimens that were either karyotypically normal individuals or patients lacking monosomy 7 and loss of 7q. The orange and green signal patterns of nuclei for each of the 25 specimens were evaluated by 2 technologists. Each technologist evaluated 100 nuclei per specimen for a total of 200 nuclei per specimen and 5000 scoreable nuclei for each of the specimen types.
The Vysis D7S486/CEP 7 FISH Probe Kit has an analytical sensitivity of 98.1% for bone marrow and 98.5% for peripheral blood.

Verification of Upper Reference Limit:
The Vysis D7S486/CEP 7 assay was performed on interphase nuclei from 25 bone marrow and 25 peripheral blood specimens from either karyotypically normal individuals or patients lacking monosomy 7 and loss of 7q. The signal patterns of 200 nuclei for each specimen type were evaluated by each of 2 technologists scoring 100 nuclei per specimen.
Among the 25 karyotypically normal specimens for both peripheral blood and bone marrow, none produced 1R1G and 1R2G signals above the 4.5% and 6.5% upper reference limits.

Reproducibility (Site-to-Site):
Two replicates of the assay were run on 2 high-positive, 2 low-positive, and 2 negative panel members at 3 sites on 5 non-consecutive days for each specimen type.
All 3 sites demonstrated 100% agreement with the known status of the negative and high positive panel members for both specimen types and both signal patterns. For the bone marrow specimens, the low positives demonstrated 88% (del(7q)) and 97% (monosomy 7) agreement. For the peripheral blood, the low positives demonstrated 95% (del(7q)) and 93% (monosomy 7) agreement.

Reproducibility (Lot-to-Lot):
Using the same panel members from the site-to-site study, 4 replicates of the assay were run on 2 high-positive, 2 low-positive, and 2 negative panel members using 3 different lots of probe at a single site for each specimen type.
All 3 lots demonstrated 100% agreement with the know status of the negative and high positive panel members for both specimen types and both signal patterns. For the bone marrow specimens, the low positives demonstrated 88% (del(7q)) and 92% (monosomy 7) agreement. For the peripheral blood, the low positives demonstrated 100% (del(7q)) and 96% (monosomy 7) agreement.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Analytical Specificity: 100%
Analytical Sensitivity (Bone Marrow): 98.1%
Analytical Sensitivity (Peripheral Blood): 98.5%

Site-to-Site Reproducibility (Bone Marrow, Low Positive del(7q)): 88% agreement
Site-to-Site Reproducibility (Bone Marrow, Low Positive monosomy 7): 97% agreement
Site-to-Site Reproducibility (Peripheral Blood, Low Positive del(7q)): 95% agreement
Site-to-Site Reproducibility (Peripheral Blood, Low Positive monosomy 7): 93% agreement

Lot-to-Lot Reproducibility (Bone Marrow, Low Positive del(7q)): 88% agreement
Lot-to-Lot Reproducibility (Bone Marrow, Low Positive monosomy 7): 92% agreement
Lot-to-Lot Reproducibility (Peripheral Blood, Low Positive del(7q)): 100% agreement
Lot-to-Lot Reproducibility (Peripheral Blood, Low Positive monosomy 7): 96% agreement

Predicate Device(s)

K123951

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 864.1870 Early growth response 1 (EGR1) gene fluorescence in-situ hybridization (FISH) test system for specimen characterization.

(a)
Identification. An early growth response 1 (EGR1) gene fluorescence in-situ hybridization (FISH) test system for specimen characterization is a device intended to detect the EGR1 probe target on chromosome 5q in bone marrow specimens from patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). The assay results are intended to be interpreted only by a qualified pathologist or cytogeneticist. These devices do not include automated systems that directly report results without review and interpretation by a qualified pathologist or cytogeneticist. These devices also do not include any device intended for use to select patient therapy, predict patient response to therapy, or to screen for disease as well as any device with a claim for a particular diagnosis, prognosis, monitoring, or risk assessment.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must also include the following information:
(i) A detailed description of all probes included in the kit;
(ii) Purpose of each probe;
(iii) Probe molecular specificity;
(iv) Probe specificity;
(v) Probe limits;
(vi) Probe sensitivity;
(vii) Specification of required ancillary reagents, instrumentation, and equipment;
(viii) Specification of the specimen collection, processing, storage and slide preparation methods;
(ix) Specification of the assay procedure;
(x) Specification of control elements that are incorporated into the recommended testing procedures;
(xi) Specification of risk mitigation elements: Description of all additional procedures, methods, and practices incorporated into the directions for use that mitigate risks associated with testing;
(xii) Specification of the criteria for test result interpretation and reporting;
(xiii) Device analytical sensitivity data;
(xiv) Device analytical specificity data;
(xv) Device reference limit data;
(xvi) Device precision/reproducibility data;
(xvii) Device stability data to include:
(A) Real-time stability,
(B) Freeze-thaw stability,
(C) Transport and temperature stability,
(D) Post-hybridization signal stability,
(E) Photostability of probe, and
(xviii) Documentation that demonstrates the clinical validity of the device. The documentation must include data from clinical studies, a minimum of two peer-reviewed published literature references using the specific device seeking marketing clearance, or both. Documentation for the clinical studies and peer-reviewed published literature references cited must include the following elements:
(A) Documentation that the sponsor's probe was used in the literature reference,
(B) Number and type of specimens,
(C) Target population studied,
(D) Upper reference limit, and
(E) Range of positive probe results.
(2) Your § 809.10(b)(12) of this chapter compliant labeling must include a statement summarizing the data identified in paragraphs (b)(1)(xiii) through (xviii) of this section and a description of the studies supporting the information, including the pre-specified acceptance criteria for these performance studies, justification for the pre-specified acceptance criteria, and whether the pre-specified acceptance criteria were met.
(3) Your § 809.10 of this chapter compliant labeling must include:
(i) A warning that reads “The assay results are intended to be interpreted only by a qualified pathologist or cytogeneticist.”
(ii) A warning that reads “This device is not for high-risk uses such as selecting therapy, predicting therapeutic response or disease screening.”
(iii) A warning that reads “The use of this device for diagnosis, monitoring or risk assessment has not been established.”

Summary

0

510(k) Summary

510(k) Number - K131508

Device Name: Vysis D7S486/CEP 7 FISH Probe Kit (List No. 04N78-020)

Purpose of the Submission

The purpose of this 510(k) is to gain clearance to market the Vysis D7S486/CEP 7 FISH Probe Kit (List No. 04N78-020).

Official Correspondent to the File

Name	Dr. Nancy Bengtson
Title:	Associate Director, Global Regulatory Affairs
Telephone:	(224) 361-7087
Fax:	(847) 775-6777
Email:	Nancy.Bengtson@abbott.com
Name:	Ms. Pamela L. Swatkowski
Title:	Director, Global 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 5, 2013

Manufacturer

Abbott Molecular Inc. is the legal manufacturer of the Vysis D7S486/CEP 7 FISH Probe Kit (List No. 04N78-020).

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

1

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

Establishment Registration No .: 3005248192

Intended Use/Indications For Use

The Vysis D7S486/CEP 7 FISH Probe Kit is a device intended for specimen characterization, and detects the LSI D7S486 probe target on chromosome 7q31 and the CEP 7 probe target on chromosome 7p11.1-g11.1 in bone marrow and peripheral blood specimens from patients with acute myeloid leukemia or myelodysplastic syndrome. The assay results are intended to be interpreted by a qualified pathologist or cytogeneticist. This device is not intended for high-risk uses such as selecting therapy, predicting therapeutic response or disease screening. The use of this device for diagnosis, prognosis, monitoring or risk assessment has not been established.

Trade Name

Vysis D7S486/CEP 7 FISH Probe Kit (List No. 04N78-020)

Common Name

Vysis D7S486/CEP 7 Fluorescence In Situ Hybridization (FISH) Probe Kit

Classification

Class II

Regulation Number

21 CFR 864.1870

510(k) Summary Page 2 of 19

2

Product Code

PFG

Predicate Device(s)

81, Immunology; PDO, Vysis EGR1 FISH Probe Kit - SC (Specimen Characterization); 510(k) K123951 :

.

Comparison with Predicate Device(s)

| Predicate | Device Item Being
Compared | Similarities | Differences: |
|-----------------------------------------------------------------------------------------------------|--------------------------------------------------|--------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| For specimen
characterization and
detection of a specific
probe target on a
chromosome. | Intended Use | Same | |
| Specimen characterization
and detection of LSI
EGR1 probe target on
chromosome 5q | Indications for Use | | Specimen
characterization and
detection of LSI
D7S486 (7q31)
probe target on
chromosome 7q and
the LSI D7S486 and
CEP 7 (7p11.1-
q11.1) probe targets
on chromosome 7. |
| Acute myeloid leukemia
(AML) patients | Patient Population | Same | In addition
myelodysplastic
syndrome |
| Vysis EGR1 FISH Probe
Kit – SC (Specimen
Characterization) | Device Name | | Vysis D7S486/CEP
7 FISH Probe Kit |
| 04N37-001 | List Number | | 04N78-020 |
| PDO | PFG | | |
| LSI EGRI | Probe Target | | LSI D7S486
CEP 7 |
| Deletion | Type of atypical FISH
signal pattern detected | Same | In addition loss of
chromosome |

3

| DNA Fluorophore labeled
Probes
Vysis LSI/WCP
Hybridization buffer,
DAPI II
NP-40

20XSSC	Kit components	Same
Predicate	Device Item Being
Compared	Similarities	Differences
FISH assay method	FISH procedure	Same
6%	Upper reference limit		4.5% (monosomy 7)
6.5% (loss of 7q)
Fluorescence In Situ
Hybridization	Technology	Same
Bone Marrow	Specimen Type	Same	Addition of
Peripheral Blood
Analytical Sensitivity
Analytical Specificity
Reproducibility	Analytical
Performance	Same
Shortest dated component
24 months
24 months
12 months
36 months
60 months	Shelf Life
Kit
• Probe
• Hybridization
buffer
• DAPI II
• NP-40
• 20X SSC	Same	Probe dating
12 months

Device Description

The Vysis D7S486/CEP 7 FISH Probe Kit is for specimen characterization and detects the LSI D7S486 (7q31) probe target on chromosome 7q31 and CEP 7 probe target chromosome 7p11.1-q11.1 in bone marrow and peripheral blood specimens.

DNA Probe Description

Vysis LSI D7S486 SpectrumOrange/ CEP 7 SpectrumGreen Probes:

4

The SpectrumOrange labeled LSI D7S486 probe is approximately 308 kb in length (chr7:115983468-115675366; February 2009 Assembly UCSC Human Genome Browser).

The SpectrumGreen labeled CEP 7 probe targets the D7Z1 alpha satellite sequence at the centromere of chromosome 7.

The Vysis D7S486/CEP 7 FISH Probe Kit (List No. 04N78-020) consists of a mixture of two DNA FISH probes and four general reagents sufficient to process 20 assays.

. Vysis LSI D7S486 SpectrumOrange/ CEP 7 SpectrumGreen Probes
. Vysis LSI/WCP Hybridization Buffer
. DAPI II Counterstain
NP-40 .
. 20X SSC

Summary and Explanation of the Test

Deletion of chromosome 7q and loss of a complete chromosome 7 (monosomy 7) are recurring abnormalities in several hematologic malignancies. Vance et al2 demonstrated the Vysis LSI D7S486 SpectrumOrange/CEP 7 SpectrumGreen Probes detected deletion of the LSI D7S486 probe target on chromosome 7q or loss of chromosome 7 (monosomy 7) in a series of bone marrow and blood specimens from untreated acute myeloid leukemia (AML) patients. In the study, 179 bone marrow specimens and 47 peripheral blood specimens were tested. Among the 179 bone marrow specimens tested, 4 bone marrow specimens were atypical for the 1R2G FISH pattern associated with a deletion of the locus specific identifier (LSI) D7S486 probe target on chromosome 7q with a range of 58 to 93% atypical nuclei. Among the 179 bone marrow specimens tested. 5 bone marrow specimens were atypical for the 1R1G FISH pattern associated with monosomy 7 with a range of 70 to 96% atypical nuclei. Among the 47 peripheral blood specimens tested, 1 peripheral blood specimen contained the 1R1G atypical FISH pattern in 11% of nuclei . An additional 3 peripheral blood specimens were atypical for the 1R2G FISH pattern with a range of 37 - 96% atypical nuclei. Cherry et als reported the Vysis LSI D7S486

5

SpectrumOrange/CEP 7 SpectrumGreen Probes detected 7g- in 3 myelodysplastic syndrome (MDS) patients with a range of 22.5 to 44% atypical bone marrow nuclej and also detected monosomy 7 in 2 patients with 23 and 87.5% atypical nuclei. Tefferi et al1 described 2 myelofibrosis with myeloid metaplasia (MMM) patients who had 9% and 16% bone marrow nuclei with 7q- (1R2G) atypical signal patterns. A peripheral blood specimen from the MMM patient with 16% atypical bone marrow nuclei had 27% of nuclei with the IR2G atypical signal pattern. The peripheral blood specimen from the patient with 9% 1R2G bone marrow was determined to be typical. However, the author stated that the percent of atypical nuclei may have been below detection limits in this peripheral blood specimen.

The Vysis D7S486/CEP 7 FISH probe kit uses FISH DNA probe technology to detect the probe target for LSI D7S496 (7g31) on chromosome 7q and the probe target for CEP 7 (7p11.1 – q11.1) on chromosome 7.

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.5

Bone marrow and peripheral blood cell specimens 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 probes of the Vysis D7S486/CEP 7 FISH Probe Kit. Following hybridization, the unbound probe is removed by a series of washes, and the nuclei are counterstained with DAPI II, a DNAspecific stain that fluoresces blue. Hybridization of the Vysis LSI D7S486 SpectrumOrange/CEP 7 SpectrumGreen Probes is viewed using a fluorescence

6

microscope equipped with appropriate excitation and emission filters, allowing visualization of the orange and green fluorescent signals.

In a cell with typical copy numbers of the Vysis LSI D7S486 SpectrumOrange/CEP 7 SpectrumGreen probe targets, two orange (2R) signals (D7S486) and two green (2G) signals (CEP 7) will be expected.

A 1 orange, 1 green (1R1G) pattern is expected in a cell having only one copy of chromosome 7. A 1 orange, 2 green (1R2G) pattern is expected in a cell with loss of 7q.

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.56 The analytical specificity of the Vysis LSI D7S486 SpectrumOrange//CEP 7 SpectrumGreen Probes for their respective chromosome target loci was established using metaphase chromosomes prepared from peripheral blood cultures of 4 male and 1 female karyotypically normal specimen slides. The hybridization location of each FISH signal on chromosomes of 100 consecutive metaphase nuclei was evaluated by 1 technologist for a total of 200 target loci per probe.

For each probe, 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.

For each probe, the specificity 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 D7S486/CEP 7 FISH Probe Kit was 100%, as shown in Table 1.

7

Table 1. Analytical Specificity
		No. of Metaphase Chromosome
Signals
Probe	Correct
Target
Locus	Hybridized to the
Correct Target
Locus	Total
Hybridized
Signals	Specificity
(%)	95%
Confidence
Interval (%)
LSI D7S486	7q31	200	200	100	(98,100)
CEP 7	7p11.1-q11.1	200	200	100	(98,100)

Analytical Sensitivity

Analytical sensitivity is defined as the percentage of scoreable interphase nuclei with the expected typical signal pattern. The expected typical interphase signal pattern for the probes in the Vysis D7S486/CEP 7 FISH Probe Kit is 2 orange (2R) and 2 green (2G) signals per nucleus.

The analytical sensitivity of the Vysis LSI D7S486 SpectrumOrange//CEP 7 SpectrumGreen Probes was established using interphase nuclei prepared from 25 bone marrow and 25 peripheral blood specimens that were either karyotypically normal individuals or patients lacking monosomy 7 and loss of 7q. The orange and green signal patterns of nuclei for each of the 25 specimens were evaluated by 2 technologists. Each technologist evaluated 100 nuclei per specimen for a total of 200 nuclei per specimen and 5000 scoreable nuclei for each of the specimen types.

The analytical sensitivity was calculated as the percentage of scoreable interphase nuclei with the expected 2R2G signal pattern.

The Vysis D7S486/CEP 7 FISH Probe Kit has an analytical sensitivity of 98.1% for bone marrow and 98.5% for peripheral blood as shown in Table 2.

8

Table 2. Analytical Sensitivity
		No. of Interphase
Chromosome Signals		Analytical Sensitivity
Probe	Specimena	With
Expected
Signal
Pattern	Scoreable
Nuclei	Point
Estimate	95% CIb
D7S486	BMb	4903	5000	98.1	(97.6,98.4)
D7S486	PBb	4923	5000	98.5	(98.1,98.8)

4 25 Karyotypically normal specimens for both specimen types.

b BM: Bone Marrow; PB: Peripheral Blood; CI: Confidence interval

Verification of Upper Reference Limit

The upper reference limit is defined as the maximum quantity of scoreable interphase nuclei with a specific atypical signal pattern at which a specimen is considered karyotypically normal for that signal pattern. The upper reference limit is expressed in terms of a percentage or the actual number of a specific atypical nuclear FISH signal pattern per the standard number of nuclei tested.

The upper reference limit for monosomy 7 is 4.5% or 9 1R1G patterns per 200 scoreable interphase nuclei, and the upper reference limit for loss of 7q is 6.5% or 13 1R2G patterns per 200 scoreable interphase nuclei. Specimens exceeding 9 1R1G patterns and/or 13 l R2G patterns per 200 scoreable nuclei are considered atypical for monosomy 7 and/or loss of q arm on chromosome 7 with the Vysis D7S486/CEP 7 FISH probe target.

The Vysis D7S486/CEP 7 assay was performed on interphase nuclei from 25 bone marrow and 25 peripheral blood specimens from either karyotypically normal individuals or patients lacking monosomy 7 and loss of 7q. The signal patterns of 200 nuclei for each specimen type were evaluated by each of 2 technologists scoring 100 nuclei per specimen.

Among the 25 karyotypically normal specimens for both peripheral blood and bone marrow, none produced 1R1G and 1R2G signals above the 4.5% and 6.5% upper reference limits.

9

Reproducibility

Two replicates of the assay were run on 2 high-positive, 2 low-positive, and 2 negative panel members at 3 sites on 5 non-consecutive days for each specimen type. The positive panel members for the site-to-site study were obtained by mixing positive cells with normal cells, for each of the specimen types, to obtain the desired levels of positivity. Results shown in Tables 3 through 6 show the overall agreement with the negative/positive status of the test panel members.

All 3 sites demonstrated 100% agreement with the known status of the negative and high positive panel members for both specimen types and both signal patterns. For the bone marrow specimens, the low positives demonstrated 88% (del(7q)) and 97% (monosomy 7) agreement. For the peripheral blood, the low positives demonstrated 95% (del(7q)) and 93% (monosomy 7) agreement.

Table 3. Overall Agreement, Site-to-Site - For Specimen type BM and signal Del
(7q)

and the control of the control of the country of the country of the country of

	Number
Category	Agreea	Disagreeb	Total	Percent Agreement
Negative	60	0	60	100%
Low Positive	53	7	60	88%
High Positive	60	0	60	100%

ª Agree is number of concordant slides.

b Disagree is number of discordant slides.

Table 4. Overall Agreement, Site-to-Site - For Specimen type BM and signal Monosomy 7 ·

	Number
Category	Agreea	Disagreeb	Total	Percent Agreement
Negative	60	0	60	100%
Low Positive	58	2	60	97%
High Positive	60	0	60	100%

ª Agree is number of concordant slides.

b Disagree is number of discordant slides.

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Table 5. Overall Agreement, Site-to-Site – For Specimen type PB and signal Del (7q)

	Number
Category	Agreea	Disagreeb	Total	Percent Agreement
Negative	60	0	60	100%
Low Positive	57	3	60	95%
High Positive	60	0	60	100%

4 Agree is number of concordant slides.

b Disagree is number of discordant slides.

Table 6. Overall Agreement, Site-to-Site - For Specimen type PB and signal Monosomy 7

	Number
Category	Agreea	Disagreeb	Total	Percent Agreement
Negative	60	0	60	100%
Low Positive	56	4	60	93%
High Positive	60	0	60	100%

ª Agree is number of concordant slides.

b Disagree is number of discordant slides.

The mean and the standard deviation (SD) of the percentage of cells with the 1R2G and

1R1G signal patterns were calculated.

11

The analysis of variance components for the site-to-site study is shown in Tables 7 through 10.

| and signal Del (7q) | | | Within-
Day
(Comp.) | Between-
Day
(Comp.) | Between-
Site
(Comp.) | Total |
|---------------------|----|------|---------------------------|----------------------------|-----------------------------|-------|
| Sample | N | Mean | SD | SD | SD | SD |
| Negative 1 | 30 | 0.5 | 0.27 | 0.32 | 0.06 | 0.43 |
| Negative 2 | 30 | 0.4 | 0.38 | 0.14 | 0.24 | 0.47 |
| Low Positive 1 | 30 | 10.5 | 2.11 | 1.41 | 1.14 | 2.78 |
| Low Positive 2 | 30 | 10.0 | 3.19 | 0.00 | 2.33 | 3.95 |
| High Positive 1 | 30 | 43.9 | 6.76 | 0.00 | 7.62 | 10.19 |
| High Positive 2 | 30 | 42.0 | 4.61 | 0.00 | 5.44 | 7.13 |

Table 7. Site-to-Site Analysis of Variance Components – For Specimen type BM and signal Del (7g)

Table 8. Site-to-Site Analysis of Variance Components – For Specimen type BM and signal Monosomy 7

| | | | Within-
Day
(Comp.) | Between-
Day
(Comp.) | Between-
Site
(Comp.) | Total |
|-----------------|----|------|---------------------------|----------------------------|-----------------------------|-------|
| Sample | N | Mean | SD | SD | SD | SD |
| Negative 1 | 30 | 0.5 | 0.67 | 0.14 | 0.49 | 0.84 |
| Negative 2 | 30 | 0.3 | 0.37 | 0.00 | 0.25 | 0.44 |
| Low Positive 1 | 30 | 8.9 | 2.61 | 0.00 | 1.19 | 2.87 |
| Low Positive 2 | 30 | 9.3 | 2.43 | 0.00 | 0.00 | 2.43 |
| High Positive 1 | 30 | 48.3 | 6.20 | 1.30 | 8.06 | 10.25 |
| High Positive 2 | 30 | 43.9 | 3.78 | 3.97 | 4.19 | 6.90 |

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Sample	N	Mean	Within-Day (Comp.) SD	Between-Day (Comp.) SD	Between-Site (Comp.) SD	Total SD
Negative 1	30	0.4	0.53	0.27	0.38	0.71
Negative 2	30	0.4	0.42	0.25	0.00	0.49
Low Positive 1	30	10.4	2.46	0.00	2.03	3.19
Low Positive 2	30	12.5	2.57	0.00	0.38	2.60
High Positive 1	30	42.1	3.26	2.05	5.82	6.98
High Positive 2	30	52.8	4.07	1.82	2.15	4.95

Table 9. Site-to-Site Analysis of Variance Components - For Specimen type PB and signal Del (7q)

Table 10. Site-to-Site Analysis of Variance Components – For Specimen type PB and signal Monosomy 7

| | | | Within-
Day
(Comp.) | Between-
Day
(Comp.) | Between-
Site
(Comp.) | Total |
|-----------------|----|------|---------------------------|----------------------------|-----------------------------|-------|
| Sample | N | Mean | SD | SD | . SD | SD |
| Negative 1 | 30 | 0.3 | 0.65 | 0.00 | 0.32 | 0.72 |
| Negative 2 | 30 | 0.4 | 0.47 | 0.20 | 0.25 | 0.57 |
| Low Positive 1 | 30 | 9.1 | 2.61 | 0.00 | 0.55 | 2.67 |
| Low Positive 2 | 30 | 6.9 | 2.35 | 0.52 | 0.27 | 2.42 |
| High Positive 1 | 30 | 44.8 | 3.77 | 0.00 | 5.35 | 6.55 |
| High Positive 2 | 30 | 38.8 | 3.90 | 3.17 | 1.63 | 5.29 |

Using the same panel members from the site-to-site study, 4 replicates of the assay were run on 2 high-positive, 2 low-positive, and 2 negative panel members using 3 different lots of probe at a single site for each specimen type. The overall agreement with the known negative/positive status of the test panel members is shown in Tables 11 through 14.

13

All 3 lots demonstrated 100% agreement with the know status of the negative and high positive panel members for both specimen types and both signal patterns. For the bone marrow specimens, the low positives demonstrated 88% (del(7q)) and 92% (monosomy 7) agreement. For the peripheral blood, the low positives demonstrated 100% (del(7q)) and 96% (monosomy 7) agreement.

	Table 11. Overall Agreement, Lot-to-Lot - For Specimen type BM and signal Del
(7q)

Category	Number			Percent
	Agreea	Disagreeb	Total	Agreement (%)
Negative	24	0	24	100%
Low Positive	21	3	24	88%
High Positive	24	0	24	100%

ª Agree is number of concordant slides.

b Disagree is number of discordant slides

Table 12. Overall Agreement, Lot-to-Lot – For Specimen type BM and signal
Monosomy 7

Category	Number			Percent
	Agreea	Disagreeb	Total	Agreement (%)
Negative	24	0	24	100%
Low Positive	22	2	24	92%
High Positive	24	0	24	100%

4 Agree is number of concordant slides.

b Disagree is number of discordant slides

Table 13. Overall Agreement, Lot-to-Lot - For Specimen type PB and signal Del (7q)

Category	Number			Percent
	Agreea	Disagreeb	Total	Agreement (%)
Negative	24	0	24	100%
Low Positive	24	0	24	100%
High Positive	24	0	24	100%

Agree is number of concordant slides.

b Disagree is number of discordant slides

14

Table 14. Overall Agreement, Lot-to-Lot - For Specimen Type PB and signal Monosomy 7

		Number
Category	Agreea	Disagreeb	Total	Percent
Agreement (%)
Negative	24	0	24	100%
Low Positive	23	1	24	96%
High Positive	24	0	24	100%

ª Agree is number of concordant slides.

b Disagree is number of discordant slides

, --

The analysis of variance components for the lot-to-lot study is shown in Tables 15 through 18.

| Table 15. Lot-to-Lot Analysis of Variance Components - For Specimen type BM

and signal Del (7q)
			Within-Lot
(Comp)	Between-Lot
(Comp)	Total
Sample	N	Mean	SD	SD	SD
Negative 1	12	0.1	0.22	0.06	0.23
Negative 2	12	0.2	0.28	0.00	0.28
Low Positive 1	12	8.7	2.31	1.63	2.82
Low Positive 2	12	11.3	1.58	0.00	1.58
High Positive 1	12	47.8	5.65	2.49	6.17
High Positive 2	12	48.9	3.53	2.66	4.42

15

| | | | Within-Lot
(Comp) | Between-Lot
(Comp) | Total |
|-----------------|----|------|----------------------|-----------------------|-------|
| Sample | N | Mean | SD | SD | SD |
| Negative 1 | 12 | 0.1 | 0.22 | 0.06 | 0.23 |
| Negative 2 | 12 | 0.0 | 0.14 | 0.00 | 0.14 |
| Low Positive 1 | 12 | 6.8 | 2.58 | 0.88 | 2.73 |
| Low Positive 2 | 12 | 8.1 | 1.16 | 0.00 | 1.16 |
| High Positive 1 | 12 | 43.9 | 6.55 | 4.86 | 8.16 |
| High Positive 2 | 12 | 38.5 | 4.03 | 3.71 | 5.48 |

Table 16. Lot-to-Lot Analysis of Variance Components – For Specimen type BM and signal Monosomy 7

Table 17. Lot-to-Lot Analysis of Variance Components - For Specimen type PB and signal Del (7q)

| | | | Within-Lot
(Comp) | Between-Lot
(Comp) | Total |
|-----------------|----|------|----------------------|-----------------------|-------|
| Sample | N | Mean | SD | SD | SD |
| Negative 1 | 12 | 0.1 | 0.22 | 0.06 | 0.23 |
| Negative 2 | 12 | 0.1 | 0.20 | 0.00 | 0.20 |
| Low Positive 1 | 12 | 11.6 | 2.14 | 0.98 | 2.35 |
| Low Positive 2 | 12 | 11.0 | 1.99 | 0.00 | 1.99 |
| High Positive 1 | 12 | 46.5 | 2.47 | 1.52 | 2.89 |
| High Positive 2 | 12 | 58.1 | 3.38 | 0.00 | 3.38 |

: : : .

16

| | | | Within-Lot
(Comp) | Between-Lot
(Comp) | Total |
|-----------------|----|------|----------------------|-----------------------|-------|
| Sample | N | Mean | SD | SD | SD |
| Negative 1 | 12 | 0.0 | 0.00 | 0.00 | 0.00 |
| Negative 2 | 12 | 0.0 | 0.14 | 0.00 | 0.14 |
| Low Positive 1 | 12 | 9.1 | 2.05 | 1.76 | 2.70 |
| Low Positive 2 | 12 | 5.7 | 1.22 | 0.00 | 1.22 |
| High Positive 1 | 12 | 37.1 | 2.87 | 0.90 | 3.01 |
| High Positive 2 | 12 | 32.0 | 3.42 | 1.69 | 3.81 |

Table 18. Lot-to-Lot Analysis of Variance Components – For Specimen type PB and signal Monosomy 7

·

.

17

Summary of Results from Cited Published Literature

Cited published literature may discuss device uses that have not been approved or cleared by FDA.

| Literature
Reference | Population
Studied | Number
and Type of
Specimens | Device Used | Observed D7S486/CEP 7 Results |
|-------------------------|-----------------------|-----------------------------------------------------------|-------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Vance et al1 | AMLa | 179 bone
marrow and
47 blood
specimensb | Vysis LSI
D7S486/CEP 7
probes | Overall 7q- (1R2G signal pattern)
was detected in 4/179 bone marrow
specimens and 3/47 peripheral blood
specimens.
Overall monosomy 7 (IR1G signal
pattern) was detected in 5/179 bone
marrow specimens and 1/47
peripheral blood specimens. |
| Cherry et al2 | MDSa | 48 bone
marrow
specimens | Vysis LSI
D7S486/CEP 7
probes | Overall 7q- (1R2G signal pattern)
was detected in 3/48 bone marrow
specimens.
Overall monosomy 7 (IRIG signal
pattern) was detected in 2/48 bone
marrow specimens. |
| Tefferi et al3 | MMMa | 42 bone
marrow and
peripheral
blood
specimens | Vysis LSI
D7S486/CEP 7
probes | Overall 7q- (1R2G signal pattern)
was detected in 2/42 bone marrow
specimens and 1/42 peripheral blood
specimens. |

Data from supporting literature

ª AML: acute myeloid leukemia; MDS: myelodysplastic syndrome; MMM: myelofibrosis with myeloid metaplasia

b Based on unpublished data.

18

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 [April 30, 2013].
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 Ttrial (E1900). Leuk Res. 2007;31:605-09.
1. Cherry A, Brockman S, Paternoster, S et al. Comparison of interphase FISH and metaphse cytogenetics to study myelodysplastic syndrome: an Eastern Cooperative Oncology Group (ECOG) study. Leuk Res. 2003;27:1085-1090.
1. Tefferi A, Meyer R. Wyatt W, et al. Comparison of peripheral blood interphase cytogenetics with bone marrow karyotype analysis in myelofibrosis with myeloid metaplasia. Br J Haematol. 2001;115:316-319.
1. Wiktor AE, Van Dyke DL, Stupca PJ et al. Preclinical validation of fluorescence in situ hybridization assays for clinical practice. Genet Med. 2006;8:16-23.
1. AMERICAN COLLEGE OF MEDICAL GENETICS (ACMG). Standards and Guidelines for Clinical Genetics Laboratories. 2008 Edition.

19

Image /page/19/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is circular and contains the words "U.S. Department of Health & Human Services" around the perimeter of the circle. Inside the circle is an abstract symbol that resembles a stylized human figure.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - W() bookings Silver Spring, MD 20993-0002

September 13. 2013

ABBOTT MOLECULAR, INC. NANCY W. BENGTSON MANAGER. GLOBAL REGULATORY AFFAIRS 1300 E. TOUHY AVENUE DES PLAINES. IL 60018

Rc: K131508

Trade/Device Name: Vysis D7S486/CEP 7 FISH Probe Kit Regulation Number: 21 CFR 864.1870 Regulation Name: Early growth response 1 (EGFRI) gene fluorescence in-situ hybridization (FISH) test system for specimen characterization Regulatory Class: II Product Code: PFG Dated: August 13, 2013 Received: August 14, 2013

Dear Dr. 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. Please note: CDRI does not evaluate information related to contract fiability 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 Parts 801 and 809); medical device reporting of medical device-related adverse events) (21 CFR 803): good manufacturing practice requirements as set 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.

20

Page 2-Dr. Bengtson

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://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/ReportalProblem/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/MedicalDevices/Resourcesfor You/Industry/default.hum.

Sincerely yours,

Reena Philip -S

for

Maria M. Chan, Ph.D.

Director, Division of Immunology and Hematology Devices

Office of In Vitro Diagnostics and Radiological Health

Center for Devices and Radiological Health

Enclosure

21

Indications for Use

510(k) Number (if known): K131508

Device Name: Vysis D7S486/CEP 7 FISH Probe Kit

Indications For Use:

The Vysis D7S486/CEP 7 FISH Probe Kit is a device intended for specimen characterization, and detects the LSI D7S486 probe target on chromosome 7q31 and the CEP 7 probe target on chromosome 7p11.1-q11.1 in bone marrow and peripheral blood specimens from patients with acute myeloid leukemia or myelodysplastic syndrome. The assay results are intended to be interpreted by a qualified pathologist or cytogeneticist. This device is not intended for high-risk uses such as selecting therapy, predicting therapeutic response or disease screening. The use of this device for diagnosis, prognosis, monitoring, or risk assessment has not been established.

Prescription Use × (Part 21 CFR 801 Subpart D)

:

AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH; Office of In Vitro Diagnostics and Radiological Health (OlR) Donna M. Roscoe:-S 2013.09.13 13:23:06 -04'00'

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k)

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