(112 days)
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.
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
The Vysis D7S486/CEP 7 FISH Probe Kit is designed for specimen characterization, specifically detecting 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.
Here's an analysis of the acceptance criteria and the study that proves the device meets them:
1. Table of Acceptance Criteria and Reported Device Performance
| Acceptance Criteria | Reported Device Performance |
|---|---|
| Analytical Specificity: Percentage of signals that hybridize to the correct locus and no other location. | LSI D7S486: 100% (95% CI: 98,100) on 7q31 |
| CEP 7: 100% (95% CI: 98,100) on 7p11.1-q11.1 | |
| Analytical Sensitivity (Bone Marrow): Percentage of scoreable interphase nuclei with the expected 2R2G signal pattern. | 98.1% (95% CI: 97.6, 98.4) |
| Analytical Sensitivity (Peripheral Blood): Percentage of scoreable interphase nuclei with the expected 2R2G signal pattern. | 98.5% (95% CI: 98.1, 98.8) |
| Upper Reference Limit (Monosomy 7): Maximum percentage of 1R1G patterns for a normal specimen (not to exceed 4.5%). | None of the 25 normal bone marrow and 25 normal peripheral blood specimens exceeded 4.5% 1R1G patterns. |
| Upper Reference Limit (Loss of 7q): Maximum percentage of 1R2G patterns for a normal specimen (not to exceed 6.5%). | None of the 25 normal bone marrow and 25 normal peripheral blood specimens exceeded 6.5% 1R2G patterns. |
| Reproducibility (Site-to-Site - Del 7q, BM): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 88% | |
| High Positive: 100% | |
| Reproducibility (Site-to-Site - Monosomy 7, BM): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 97% | |
| High Positive: 100% | |
| Reproducibility (Site-to-Site - Del 7q, PB): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 95% | |
| High Positive: 100% | |
| Reproducibility (Site-to-Site - Monosomy 7, PB): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 93% | |
| High Positive: 100% | |
| Reproducibility (Lot-to-Lot - Del 7q, BM): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 88% | |
| High Positive: 100% | |
| Reproducibility (Lot-to-Lot - Monosomy 7, BM): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 92% | |
| High Positive: 100% | |
| Reproducibility (Lot-to-Lot - Del 7q, PB): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 100% | |
| High Positive: 100% | |
| Reproducibility (Lot-to-Lot - Monosomy 7, PB): Overall agreement with negative/positive status. | Negative: 100% |
| Low Positive: 96% | |
| High Positive: 100% |
2. Sample Size Used for the Test Set and Data Provenance
- Analytical Specificity: 4 male and 1 female karyotypically normal specimen slides.
- Analytical Sensitivity and Verification of Upper Reference Limit: 25 bone marrow and 25 peripheral blood specimens. These were from either karyotypically normal individuals or patients lacking monosomy 7 and loss of 7q. The provenance is not explicitly stated (e.g., country of origin, retrospective/prospective).
- Reproducibility (Site-to-Site & Lot-to-Lot): The panel members for the reproducibility studies were prepared by mixing positive cells with normal cells. The exact number of individual patient samples from which these positive and normal cells originated is not specified. The study used 2 high-positive, 2 low-positive, and 2 negative panel members for each specimen type (bone marrow and peripheral blood). The data provenance is not specified as retrospective or prospective, nor is the country of origin explicitly mentioned.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
- Analytical Specificity: 1 technologist for evaluating metaphase chromosomes.
- Analytical Sensitivity and Verification of Upper Reference Limit: 2 technologists for evaluating interphase nuclei.
- Reproducibility: The ground truth for the reproducibility studies (negative, low positive, high positive categories) was established by mixing positive cells with normal cells to achieve desired levels of positivity, implying a predefined "known status" based on these mixtures. The expertise used to determine the initial "positive cells" or "normal cells" is not detailed.
The qualifications of these technologists/experts are not explicitly stated (e.g., years of experience, specific certifications like "radiologist with 10 years of experience").
4. Adjudication Method (e.g., 2+1, 3+1, none) for the Test Set
For Analytical Sensitivity and Verification of Upper Reference Limit, the document states that each technologist evaluated 100 nuclei per specimen, implying independent scoring. There is no mention of an adjudication method (like 2+1 or 3+1) if scores differed between the two technologists, nor is an adjudication method specified for other tests.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done, What Was the Effect Size of How Much Human Readers Improve with AI vs. Without AI Assistance?
No MRMC comparative effectiveness study was done. This device is a FISH probe kit, not an AI-assisted diagnostic tool. Therefore, there is no discussion of human reader improvement with or without AI assistance.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done?
This is a laboratory diagnostic kit (FISH probe) that requires human interpretation (a qualified pathologist or cytogeneticist). Therefore, a standalone algorithm-only performance assessment is not applicable and was not performed. The performance studies evaluate the kit's analytical characteristics.
7. The Type of Ground Truth Used (Expert Consensus, Pathology, Outcomes Data, etc.)
- Analytical Specificity: Karyotypically normal specimens were used, with the "correct locus" pre-defined based on known chromosomal locations.
- Analytical Sensitivity and Verification of Upper Reference Limit: Karyotypically normal individuals or patients lacking the specific abnormalities (monosomy 7 and loss of 7q) were used. The expected typical signal pattern (2R2G) served as the ground truth for normalcy. The "atypical" patterns (1R1G, 1R2G) were defined based on the biological expectation of monosomy 7 or 7q deletion.
- Reproducibility: The ground truth for the panel members was established by preparing mixtures of positive and normal cells to achieve predefined "negative," "low positive," and "high positive" statuses. The origin of the "positive cells" would implicitly be from samples with confirmed deletion 7q or monosomy 7, likely established through standard cytogenetic or FISH methods.
8. The Sample Size for the Training Set
This document describes a diagnostic kit and its analytical validation. It does not refer to a machine learning or AI algorithm development pipeline, so there is no specific mention of a "training set" in the context of algorithm development. The studies performed are for analytical validation.
9. How the Ground Truth for the Training Set Was Established
As there is no mention of a "training set" for an algorithm, this question is not applicable. The kit is based on established FISH technology, and its performance is validated through analytical studies.
§ 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.”