(718 days)
The ACTOnco IVD assay is an in vitro diagnostic test that uses targeted next generation sequencing of formalin-fixed. paraffin-embedded tumor tissue from patients with solid malignant neoplasms to detect genetic alterations in a broad multi gene panel. The test is intended to provide informations, small insertions and deletions, ERBB2 gene amplification, and tumor mutational burden for use by qualified health care professionals in accordance with professional guidelines, and is not conclusive or prescriptive for labeled use of any specific therapeutic product. ACTOnco IVD is a single-site assay performed at ACT Genomics.
The ACTOnco IVD assay is an in-vitro diagnostic assay intended to provide information for use by qualified health care professionals in accordance with professional guidelines, and is not conclusive or prescriptive for labeled use of any specific therapeutic product.
The assay is a custom targeted sequencing platform, utilizing amplicon-based sequencing, to detect point mutations (single nucleotide variants, or SNVs), small insertions and deletions (Indels), ERBB2 gene amplification, and tumor mutational burden (TMB) in tumor specimens. The assay uses custom DNA primers corresponding to all exons and selected introns of oncogenes, tumor suppressor genes, drug metabolism genes, and immune-related genes. Primers are synthesized by a secondary manufacturer (Thermo Fisher Scientific). An overlapping amplicon approach is utilized in which tiled primers are designed to generate multiple overlapping amplicons of the same region to avoid allele dropout. In total, the primers target approximately 1.8Mb of the human genome. Genomic DNA is extracted from FFPE tissue samples.
Sequence libraries are prepared through a multiplex polymerase chain reaction (PCR) amplification step to enrich target sequences. Target sequences are tagged with index oligonucleotide to identify individual sample and adaptor oligonucleotide to anchor the amplicon to complimentary oligonucleotides embedded on the surface of the sequencing bead. Target sequences on the sequencing beads are amplified using emulsion PCR before sequencing. Multiple barcoded sequence libraries (from different patients) are pooled and then sequenced on a Thermo Fisher Ion GeneStudio™ S5 Prime System. Sequence reads are then aligned to the reference human genome. By comparing the identity of bases from the tumor DNA and the reference human genome, variant alterations are identified in the tumor.
The assay system includes a sequencing instrument, reagents (DNA extraction, library preparation and sequencing), software (operation of the sequencing instrument and variant calling), and standard operating procedures (SOPs) for the use of the system. ACT Genomics takes the responsibilities in monitoring the instrument; reagents and consumable materials which will be used in the assay process.
Multiple software components will be used in the assay. The NGS raw read analysis will be done using Thermo Fisher software. Variant calling for SNVs, insertions and deletions will be done using Thermo Fisher software. Mutation and variant annotation will be done using software from ACT Genomics, the Cunningham Lab and Golden Helix software. ERBB2 gene amplification will be done using software from Boeva Lab. Tumor Purity and Zygosity will be done using software from Halgamuge Lab (Kaushalya Amarasinghe). Calculations for tumor mutational burden will be done using ACT Genomics software.
I will analyze the provided text to extract information about the acceptance criteria and the study proving the device meets these criteria. I will then structure this information according to your requested format.
However, based on the provided text, it appears to be a 510(k) summary for a medical device (ACTOnco IVD, a next-generation sequencing-based tumor profiling test). The document describes performance testing (precision/reproducibility, analytical sensitivity/limit of detection, analytical specificity/interference, cross-contamination, DNA input, DNA extraction) and method comparison data.
Crucially, this document
does not contain a direct table of acceptance criteria nor explicit statements about "acceptance criteria" met by the device.
It also does not describe a "Multi Reader Multi Case (MRMC) comparative effectiveness study" or information about "human readers improve with AI vs without AI assistance" as it is a diagnostic test and not an AI-assisted diagnostic imaging device/software for human readers.
Therefore, I will provide the acceptance criteria based on the performance observed in the various analytical validation studies, as these implicitly define the performance considered acceptable for the device. I will also make clear when certain information is not present in the provided text.
Here is the information structured according to your request, extracted from the provided 510(k) summary:
Device: ACTOnco IVD - Next generation sequencing based tumor profiling test
1. Table of Acceptance Criteria (Inferred from Performance Studies) and Reported Device Performance:
Since explicit acceptance criteria are not called out as a separate table, I will infer them from the "Performance Testing" sections, assuming that the reported performance metrics were considered acceptable for the device's clearance.
| Performance Metric Category & Specificity | Inferred Acceptance Criterion (Target) | Reported Device Performance (observed outcomes from studies) |
|---|---|---|
| Precision / Reproducibility (SNVs, Indels) | ||
| Overall Call Rate (Positive) | ≥ 95% | 98.33% (35,308/35,906) |
| SNV Call Rate | ≥ 95% | 98.33% (31,837/32,377) |
| MNV Call Rate | ≥ 95% | 97.18% (963/991) |
| Insertion (INS) Call Rate | ≥ 95% | 96.97% (512/528) |
| Deletion (DEL) Call Rate | ≥ 95% | 99.30% (1,996/2,010) |
| Overall Call Rate (Negative/WT) | ≥ 99% | 99.997% (723,628/723,648) |
| Precision / Reproducibility (ERBB2 Amplification) | ||
| Positive Call Rate | 100% | 100% (based on 144 observations from 3 amplified samples) |
| Negative Call Rate | 100% | 100% (based on 816 observations from 17 non-amplified samples) |
| Precision / Reproducibility (TMB) | CV% ≤ 16% for TMB scores when Tumor Purity ≥ 20% | Ranges from 4.151% to 59.144% (for tumor purity 32.5% to 72.7%). Explicitly stated acceptable across tumor purities at or above 20% with percent CV 30%. |
| Analytical Sensitivity / Limit of Detection (LoD) | ||
| SNVs (Hotspot, 2% cutoff) | LoD/C95 for individual variants evaluated. Target to be near or below the cutoff. | Established MAF Range: 1.5%-6.6% |
| SNVs (Non-hotspot, 5% cutoff) | LoD/C95 for individual variants evaluated. Target to be near or below the cutoff. | Established MAF Range: 2.4%-15.1% |
| Insertions | LoD/C95 for individual variants evaluated. Target to be near or below the cutoff. | Established MAF Range: 1.1%-45.4% |
| Deletions | LoD/C95 for individual variants evaluated. Target to be near or below the cutoff. | Established MAF Range: 1.9%-22.0% |
| TMB (DNA Input) | Acceptable CV across relevant tumor purity and DNA input ranges | Acceptable TMB performance across tumor purities at or above 20% with percent CV 30%. |
| ERBB2 Amplification (LoD) | 100% call rate for samples > 10% tumor purity | 100% call rate for samples with tumor purity over 10%. (Data supported consistent repeatability at tumor purity 30%). |
| Analytical Specificity / Interference (SNVs and Indels) | ||
| Correct Positive Calls (Spike-in) | 100% (consistent with control) | 100% (3288/3288) with lower 95% CI of 99.88% |
| Correct Negative Calls (Spike-in) | 99.99% (consistent with control) | 99.99% (122,656/122,664) with lower 95% CI of 99.99% |
| Necrotic Tissue Tolerance | Acceptable processing and agreement up to 50% necrotic tissue | Control Group ( |
§ 866.6080 Next generation sequencing based tumor profiling test.
(a)
Identification. A next generation sequencing (NGS) based tumor profiling test is a qualitative in vitro diagnostic test intended for NGS analysis of tissue specimens from malignant solid neoplasms to detect somatic mutations in a broad panel of targeted genes to aid in the management of previously diagnosed cancer patients by qualified health care professionals.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include the following information:
(i) A detailed description of all somatic mutations that are intended to be detected by the test and that are adequately supported in accordance with paragraph (b)(1)(v) of this section and reported in the test results in accordance with paragraph (b)(2)(iv) of this section, including:
(A) A listing of mutations that are cancer mutations with evidence of clinical significance.
(B) As appropriate, a listing of mutations that are cancer mutations with potential clinical significance.
(ii) The indications for use must specify the following:
(A) The test is indicated for previously diagnosed cancer patients.
(B) The intended specimen type(s) and matrix (
e.g., formalin-fixed, paraffin-embedded tumor tissue).(C) The mutation types (
e.g., single nucleotide variant, insertion, deletion, copy number variation or gene rearrangement) for which validation data has been provided.(D) The name of the testing facility or facilities, as applicable.
(iii) A detailed device description including the following:
(A) A description of the test in terms of genomic coverage, as follows:
(
1 ) Tabulated summary of all mutations reported, grouped according to gene and target region within each gene, along with the specific cDNA and amino acid positions for each mutation.(
2 ) A description of any within-gene targeted regions that cannot be reported and the data behind such conclusion.(B) Specifications for specimen requirements including any specimen collection devices and preservatives, specimen volume, minimum tumor content, specimen handling, DNA extraction, and criteria for DNA quality and quantity metrics that are prerequisite to performing the assay.
(C) A detailed description of all test components, reagents, instrumentation, and software required. Detailed documentation of the device software including but not limited to, software applications and hardware-based devices that incorporate software.
(D) A detailed description of the methodology and protocols for each step of the test, including description of the quality metrics, thresholds, and filters at each step of the test that are implemented for final result reporting and a description of the metrics for run-failures, specimen-failures, invalids, as applicable.
(E) A list of links provided by the device to the user or accessed by the device for internal or external information (
e.g., decision rules or databases) supporting clinical significance of test results for the panel or its elements in accordance with paragraphs (b)(1)(v) and (b)(2)(vi) of this section.(F) A description of internal and external controls that are recommended or provided and control procedures. The description must identify those control elements that are incorporated into the testing procedure.
(iv) Information demonstrating analytical validity of the device according to analytical performance characteristics, evaluated either specifically for each gene/mutation or, when clinically and practically justified, using a representative approach based on other mutations of the same type, including:
(A) Data that adequately supports the intended specimen type (
e.g., formalin-fixed, paraffin-embedded tumor tissue), specimen handling protocol, and nucleic acid purification for specific tumor types or for a pan-tumor claim.(B) A summary of the empirical evidence obtained to demonstrate how the analytical quality metrics and thresholds were optimized.
(C) Device precision data using clinical samples to adequately evaluate intra-run, inter-run, and total variability. The samples must cover all mutation types tested (both positive and negative samples) and include samples near the limit of detection of the device. Precision must be assessed by agreement within replicates on the assay final result for each representative mutation, as applicable, and also supported by sequencing quality metrics for targeted regions across the panel.
(D) Description of the protocols and/or data adequately demonstrating the interchangeability of reagent lots and multiplexing barcodes.
(E) A description of the nucleic acid assay input concentration range and the evidence to adequately support the range.
(F) A description of the data adequately supporting the limit of detection of the device.
(G) A description of the data to adequately support device accuracy using clinical specimens representing the intended specimen type and range of tumor types, as applicable.
(
1 ) Clinical specimens tested to support device accuracy must adequately represent the list of cancer mutations with evidence of clinical significance to be detected by the device.(
2 ) For mutations that are designated as cancer mutations with evidence of clinical significance and that are based on evidence established in the intended specimen type (e.g., tumor tissues) but for a different analyte type (e.g., protein, RNA) and/or a measurement (e.g., incorporating a score or copy number) and/or with an alternative technology (e.g., IHC, RT-qPCR, FISH), evidence of accuracy must include clinically adequate concordance between results for the mutation and the medically established biomarker test (e.g., evidence generated from an appropriately sized method comparison study using clinical specimens from the target population).(
3 ) For qualitative DNA mutations not described in paragraph (b)(1)(iv)(G)(2 ) of this section, accuracy studies must include both mutation-positive and wild-type results.(H) Adequate device stability information.
(v) Information that adequately supports the clinical significance of the panel must include:
(A) Criteria established on what types and levels of evidence will clinically validate a mutation as a cancer mutation with evidence of clinical significance versus a cancer mutation with potential clinical significance.
(B) For representative mutations of those designated as cancer mutations with evidence of clinical significance, a description of the clinical evidence associated with such mutations, such as clinical evidence presented in professional guidelines, as appropriate, with method comparison performance data as described in paragraph (b)(1)(iv)(G) of this section.
(C) For all other mutations designated as cancer mutations with potential clinical significance, a description of the rationale for reporting.
(2) The 21 CFR 809.10 compliant labeling and any product information and test report generated, must include the following, as applicable:
(i) The intended use statement must specify the following:
(A) The test is indicated for previously diagnosed cancer patients.
(B) The intended specimen type(s) and matrix (
e.g., formalin-fixed, paraffin-embedded tumor tissue).(C) The mutation types (
e.g., single nucleotide variant, insertion, deletion, copy number variation or gene rearrangement) for which validation data has been provided.(D) The name of the testing facility or facilities, as applicable.
(ii) A description of the device and summary of the results of the performance studies performed in accordance with paragraphs (b)(1)(iii), (b)(1)(iv), and (b)(1)(v) of this section.
(iii) A description of applicable test limitations, including, for device specific mutations validated with method comparison data to a medically established test in the same intended specimen type, appropriate description of the level of evidence and/or the differences between next generation sequencing results and results from the medically established test (
e.g., as described in professional guidelines).(iv) A listing of all somatic mutations that are intended to be detected by the device and that are reported in the test results under the following two categories or equivalent designations, as appropriate: “cancer mutations panel with evidence of clinical significance” or “cancer mutations panel with potential clinical significance.”
(v) For mutations reported under the category of “cancer mutations panel with potential clinical significance,” a limiting statement that states “For the mutations listed in [cancer mutations panel with potential clinical significance or equivalent designation], the clinical significance has not been demonstrated [with adequate clinical evidence (
e.g., by professional guidelines) in accordance with paragraph (b)(1)(v) of this section] or with this test.”(vi) For mutations under the category of “cancer mutations panel with evidence of clinical significance,” or equivalent designation, link(s) for physicians to access internal or external information concerning decision rules or conclusions about the level of evidence for clinical significance that is associated with the marker in accordance with paragraph (b)(1)(v) of this section.