(185 days)
The QuantideX qPCR BCR-ABL IS Kit is an in vitro nucleic acid amplification test for the quantitation of BCR-ABL1 and ABL1 transcripts in total RNA from whole blood of diagnosed t(9,22) positive Chronic Myeloid Leukemia (CML) patients expressing BCR-ABL1 fusion transcripts type e13a2 and/or e14a2. The QuantideX qPCR BCR-ABL IS Kit is a reverse transcription-quantitative PCR performed on the Applied Biosystems 7500 Fast Dx Real-Time PCR Instrument and is intended to measure BCR-ABL1 to ABL1, expressed as a log molecular reduction (MR value) from a baseline of 100% on the International Scale, in t(9;22) positive CML patients during monitoring of treatment with Tyrosine Kinase Inhibitors (TKIs).
The test does not differentiate between e13a2 or e14a2 fusion transcripts and does not monitor other rare fusion transcripts resulting from t(9:22). This test is not intended for the diagnosis of CML.
The QuantideX qPCR BCR-ABL IS Kit reagents are adapted for use on the ABI 7500 Fast Dx Real-Time PCR Instrument. The assay includes reagents sufficient for 60 reactions. A description of the reagents provided is described below in Table 1.
Here's a summary of the acceptance criteria and study that proves the device meets them, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document describes both analytical and clinical performance acceptance criteria.
Analytical Performance - Precision/Reproducibility
| Performance Metric | Acceptance Criteria (SD) | Reported Device Performance (Max Total SD) | Met? |
|---|---|---|---|
| Within-lab Precision | |||
| MR Value 4.25 | ≤ 0.36 | 0.134 (at MR 4) | Yes |
| Site-to-Site Reproducibility | |||
| MR Value 4.25 | ≤ 0.36 | 0.167 (at MR 4) | Yes |
Analytical Performance - Linearity
| Performance Metric | Acceptance Criteria (Slope/Intercept for Regression) | Reported Device Performance (Slope/Intercept/Max SD) | Met? |
|---|---|---|---|
| Linear Regression | Slope close to 1 (not explicitly defined but implied typical regression expectations), intercept close to 0 (implied) | e13a2: Slope 1.01, Intercept -0.11, Max SD 0.17 | |
| e14a2: Slope 1.01, Intercept -0.05, Max SD 0.17 | Yes | ||
| Range of Linearity | (Implied based on clinical use range) | MR 0.3 (50% IS) to MR 4.7 (0.002% IS) for both transcripts | Yes |
Analytical Performance - Limit of Quantitation (LoQ)
| Performance Metric | Acceptance Criteria (SD) | Reported Device Performance (MR Range/SD Range) | Met? |
|---|---|---|---|
| LoQ | ≤ 0.36 at MR 4.5 or greater | MR values 4.6 to 4.87, SD values 0.23 to 0.34 | Yes |
Analytical Performance - Interfering Substances
| Performance Metric | Acceptance Criteria (Mean difference from control) | Reported Device Performance | Met? |
|---|---|---|---|
| Interference | Mean of test samples ± 0.5 MR of the control | All met acceptance criteria | Yes |
Analytical Performance - Primer Specificity
| Performance Metric | Acceptance Criteria | Reported Device Performance | Met? |
|---|---|---|---|
| Specificity | Specimens without a major BCR-ABL1 breakpoint reported as negative or below LoD in > 8/9 replicates | Met acceptance criteria (e.g., cell lines and IVT controls without e13a2/e14a2 were 95% of CML-negative samples tested negative or below LoD | All negative specimens (25/25) reported as "Negative (sufficient ABL1)" |
Analytical Performance - RNA Input
| Performance Metric | Acceptance Criteria (SD criteria from Table 3) | Reported Device Performance (RNA input range with met criteria) | Met? |
|---|---|---|---|
| RNA Input Range | Same as Table 3 for corresponding MR values | 750 to 6000 ng from MR 1 to MR 4.5 | Yes |
Analytical Performance - Traceability
| Performance Metric | Acceptance Criteria (Regression to WHO panel) | Reported Device Performance (Slope/Intercepts for 3 lots) | Met? |
|---|---|---|---|
| Traceability | Slope 1.0 to 1.1, Intercept 0.02 - 0.11 | Lot1: y=-0.11+1.1x; Lot2: y=0.02+1x; Lot3: y=-0.059+1.1x | Yes |
Analytical Performance - Specimen Stability (Whole Blood)
| Performance Metric | Acceptance Criteria (MR unit difference from baseline) | Reported Device Performance (Duration and storage conditions) | Met? |
|---|---|---|---|
| Whole Blood Stability | All results within 0.5 MR units from baseline | Up to 72 hours at 2-8°C | Yes |
Clinical Performance
| Performance Metric | Acceptance Criteria | Reported Device Performance | Met? |
|---|---|---|---|
| Event-Free Survival (EFS) Difference at 36 months | Statistically significantly different AND point estimates differing by at least 10 percentage points (for MR 0 | Met | Yes |
2. Sample Size Used for the Test Set and Data Provenance
The primary test set for clinical performance involved a retrospective study with:
- Sample Size: 137 evaluable samples from 96 subjects (out of 139 samples from 98 patients initially collected).
- Data Provenance: Retrospective, collected at 2 clinical sites in the US (OHSU and Hospital of the University of Pennsylvania).
For analytical performance studies, various sample compositions and quantities were used:
- Precision (within-lab & site-to-site): 25 samples formulated from 5 human RNA specimens positive for BCR-ABL1 diluted into RNAs from CML-negative blood. The site-to-site reproducibility study involved 1200 measurements from this 25-sample pool.
- Linearity/Reportable Range: 2 separate RNA specimens (one e13a2, one e14a2) diluted into RNAs from CML-negative whole blood (ranging from MR 0.1 to MR 4.8).
- Limit of Blank (LoB): 30 non-leukemic human RNA specimens.
- Limit of Detection (LoD): 4 separate human RNA specimens positive for BCR-ABL1, serially diluted to 28 levels.
- Limit of Quantitation (LoQ): 6 specimens derived from 6 human RNA positive for BCR-ABL (diluted to target MR 4.7).
- Interfering Substances: Residual CML-positive blood diluted to approximately MR 4.0 into RNA from CML-negative whole blood (tested in 9 replicates).
- Primer Specificity: 11 leukemic specimens (CML, AML, ALL) and 2 non-leukemic RNA specimens.
- Specimen Carryover Contamination: Serially diluted total RNA from residual clinical CML-positive blood into RNA from CML-negative whole blood in a checkerboard pattern (25 high positive, 25 negative).
- RNA Isolation: CML-positive white blood cells serially diluted across 4 levels (MR 1-4) into CML-negative human anti-coagulated whole blood.
- RNA Input: 30 samples derived from 2 primary RNA samples diluted into non-leukemic human RNA (total 216 evaluable observations).
- Traceability: WHO Reference Panel (4 panel members tested in duplicate).
- Reagent Stability: 5 samples diluted to approximately MR 1, 2, 3, 4, and
§ 866.6060 BCR-ABL quantitation test.
(a)
Identification. A BCR-ABL quantitation test is identified as a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) test for the quantitation of BCR-ABL1 expressed on the International Scale (IS) and control transcripts in total RNA from whole blood of diagnosed t(9;22) positive chronic myeloid leukemia (CML) patients during monitoring of treatment with tyrosine kinase inhibitors. This test is not intended for the diagnosis of CML.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include the following information:
(i) The indication for use must indicate the variant(s) for which the assay was designed and validated, for example BCR-ABL e13a2 and/or e14a2.
(ii) A detailed description of all components in the test, including the following:
(A) A detailed description of the test components, all required reagents, instrumentation and equipment, including illustrations or photographs of non-standard equipment or methods;
(B) Detailed documentation of the device software including, but not limited to, standalone software applications and hardware-based devices that incorporate software;
(C) Methodology and protocols for control procedures for the assay to allow reporting on the International Scale;
(D) A description of the result outputs, analytical sensitivity of the assay, and the range of values that will be reported; and
(E) A description of appropriate internal and external controls that are recommended or provided. The description must identify those control elements that are incorporated into the testing procedure.
(iii) Information that demonstrates the performance characteristics of the test, including:
(A) For indications for use based on a threshold established in a predicate device of this generic type, device performance data from either a method comparison study to the predicate device or through a clinical study demonstrating clinical validity using well-characterized prospectively or retrospectively obtained clinical specimens, as appropriate, representative of the intended use population;
(B) For indications for use based on a threshold not established in a predicate device of this generic type, device performance data from a clinical study demonstrating clinical validity using well-characterized prospectively or retrospectively obtained clinical specimens, as appropriate, representative of the intended use population;
(C) Device reproducibility data generated, using a minimum of three sites, of which at least two sites must be external sites, with two operators at each site. Each site must conduct a minimum of three runs per operator over non-consecutive days evaluating a minimum of five different BCR-ABL concentrations that span and are well distributed over the measuring range and include MR3 (0.1 percent IS). Results shall be reported as the standard deviation and percentage coefficient of variation for each level tested. Prespecified acceptance criteria must be provided and followed;
(D) Device precision data using clinical samples to evaluate the within-lot, between-lot, within-run, between run, and total variation;
(E) Device linearity data using a dilution panel created from clinical samples;
(F) Device analytic sensitivity data, including limit of blank, limit of detection, and limit of quantification;
(G) Device specificity data, including interference and cross-contamination; and
(H) Device stability data, including real-time stability of samples under various storage times, temperatures, and freeze-thaw conditions.
(iv) Identification of risk mitigation elements used by your device, including a detailed description of all additional procedures, methods, and practices incorporated into the instructions for use that mitigate risks associated with testing using your device.
(2) Your 21 CFR 809.10 compliant labeling must include the following:
(i) The intended use in your 21 CFR 809.10(a)(2) and (b)(2) complaint labeling must include an indication for use statement that reads “This test is not intended for the diagnosis of CML”; and
(ii) A detailed description of the performance studies conducted to comply with paragraph (b)(1)(iii) of this section and a summary of the results.
(3) Your device output must include results on the International Scale (IS) and your assay must include multipoint calibration controls traceable to a relevant international reference panel (
e.g., the World Health Organization International Genetic Reference Panel for quantitation of BCR-ABL mRNA).