(254 days)
Not Found
No
The description focuses on RT-qPCR technology and standard statistical analysis methods (linear regression, probit regression, ANOVA, Deming regression, Bland-Altman analysis). There is no mention of AI or ML algorithms being used for data processing, interpretation, or decision-making.
No
This device is an in vitro diagnostic test used for monitoring CML patients' responses to treatment by quantifying certain mRNA transcripts, not for direct therapeutic intervention.
Yes
The device is described as an "in vitro diagnostic test" intended for the "quantitation of BCR-ABL1 and ABL1 mRNA transcripts in peripheral blood specimens of diagnosed t(9;22) positive Chronic Myeloid Leukemia (CML) patients." While it explicitly states it is "not intended for the diagnosis of CML," its purpose is to measure ratios and MR values "in t(9;22) positive CML patients during of treatment," which are diagnostic measurements used for monitoring disease progression and treatment efficacy.
No
The device is an in vitro diagnostic test that utilizes hardware components (Cepheid GeneXpert systems, disposable cartridges, syringe drive, thermocycler) in addition to software for performing RT-qPCR and data analysis.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the Xpert BCR-ABL Ultra test is an "in vitro diagnostic test".
- Device Description: The "Device Description" section also refers to the test as an "automated in vitro diagnostic test".
- Function: The test is designed to analyze biological specimens (peripheral blood) in vitro (outside of the body) to provide information about a patient's health status (quantitation of BCR-ABL1 and ABL1 mRNA transcripts in CML patients).
- Regulatory Context: The document mentions comparison to an "FDA-cleared molecular assay" and adherence to "CLSI guidelines", which are standard for IVD devices.
- Predicate Device: A predicate device (DEN160003; Asuragen QuantideX qPCR BCR-ABL IS Kit) is listed, which is also an IVD.
All of these points confirm that the Xpert BCR-ABL Ultra test is an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
The Xpert BCR-ABL Ultra test is an in vitro diagnostic test for the quantitation of BCR-ABL1 and ABL1 mRNA transcripts in peripheral blood specimens of diagnosed t(9;22) positive Chronic Myeloid Leukemia (CML) patients expressing BCR-ABLI fusion transcripts type e13a2 and/or e14a2. The test utilizes automated, quantitative, real-time reverse transcription polymerase chain reaction (RT-qPCR). The Xpert BCR-ABL Ultra test is intended to measure BCR-ABL1 to ABL1 percent ratios on the International Scale (IS), and also expressed as a log molecular reduction (MR value) from a baseline of 100% (IS), in t(9;22) positive CML patients during of treatment with Tyrosine Kinase Inhibitors (TKIs).
The test does not differentiate between e13a2/b2a2 or e14a2/b3a2 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 Xpert BCR-ABL Ultra test is intended for use only on the Cepheid GeneXpert® Dx System and the GeneXpert Infinity System.
Product codes (comma separated list FDA assigned to the subject device)
OYX, OOI
Device Description
The Xpert BCR-ABL Ultra test is an automated in vitro diagnostic test for quantifying the amount of BCR-ABLI (BCR-ABL, hereafter) mRNA transcript as a ratio of BCR-ABL/ABL per the International Scale (IS).
The test is performed on the Cepheid GeneXpert® Dx System and GeneXpert Infinity System (referred to as the GeneXpert systems). The GeneXpert systems require the use of single-use, disposable cartridges that hold the PCR reagents and host the PCR process. Because the cartridges are self-contained and specimens never come into contact with working parts of the instrument modules, cross-contamination between samples is minimized. The GeneXpert systems have 1 to 80 randomly accessible modules, depending upon the instrument, that are each capable of performing separate sample preparation and real-time PCR and RT-PCR tests. Each module contains a syringe drive for dispensing fluids (i.e., the syringe drive activates the plunger that works in concert with the rotary valve in the cartridge to move fluids between chambers), and a proprietary I-CORE® thermocycler for performing real-time PCR and RT-PCR and detection.
The Xpert BCR-ABL Ultra test includes reagents to detect BCR-ABL fusion genes resulting from two major breakpoints, translocation e13a2/b2a2 and e14a2/b3a2 and the ABL transcript as an endogenous control in peripheral blood specimens. The amount of BCR-ABL transcript in the patient sample is reported as a percent ratio of BCR-ABL/ABL on the International Scale (IS), and also expressed as a log molecular reduction (MR value) from a baseline of 100% (IS), using the GeneXpert software.
There are two controls included in each Xpert BCR-ABL Ultra test, which are the ABL Endogenous Control and the Probe Check Control (PCC). The ABL Endogenous Control normalizes the BCR-ABL target and ensures that sufficient sample is used in the test. The PCC verifies reagent rehydration. PCR tube filling, and that all reaction components, including probes and dyes, are present and functional in the cartridge.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
peripheral blood specimens
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Prescription Use
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)
Non-Clinical Studies:
Linearity/Dynamic Range: Evaluated independently for e13a2/b2a2 and e14a2/b3a2 breakpoints using CML clinical specimens. Panel members were tested on two assay kit lots in replicates of 4 per kit lot. Linear regression analyses were performed for first, second, and third order polynomials. The data support an observation of linearity from at least 55% (IS)/MR 0.26 to ~0.0019% (IS)/MR4.75 with a maximum SD of 0.26. The reportable range spans from the limits of linearity at 55% (IS)/MR0.26 to the LoD/LOQ at 0.0030% (IS)/MR4.52.
Analytical Sensitivity: The limit of detection (LoD) was estimated for both e13a2/b2a2 and e14a2/b3a2 breakpoints by testing serial dilutions of High CML positive specimens and Low CML positive specimens. The estimated LoD was 0.0035% (IS)/MR4.45 for e13a2/b2a2 and 0.0030% (IS)/MR4.52 for e14a2/b3a2. The overall LoD for both is 0.0030% (IS)/MR4.52. The limit of quantitation (LoQ) was determined to be equal to the LoD, 0.0030% (IS)/MR4.52. The limit of blank (LoB) was 0.00% (IS).
Analytical Specificity: Evaluated using EDTA whole blood specimens from fifty (50) healthy donors and twenty (20) leukemic specimens (AML/ALL). Breakpoint specificity was determined by testing normal healthy donor EDTA blood spiked with five (5) different leukemia cell lines. No BCR-ABL signal was detected in healthy non-CML or AML/ALL leukemic specimens. The test showed 100% analytical specificity for non-CML EDTA blood specimens.
Potentially Interfering Substances Study: Evaluated five substances that may interfere, based on CLSI document EP07-A2. No clinically significant inhibitory effects were observed.
Carry-Over Contamination: Demonstrated that single-use, self-contained GeneXpert cartridges prevent carry-over contamination. All twenty BCR-ABL positive samples were correctly reported as POSITIVE, and all twenty BCR-ABL negative samples were correctly reported as NEGATIVE.
Traceability to WHO Panel: Demonstrated traceability to the 1st World Health Organization (WHO) International Genetic Reference Panel (NIBSC code: 09/138) by measuring the WHO Reference Panel with 3 lots of the Xpert BCR-ABL Ultra test. The slope of the line was close to unity (0.96 to 1.1) and the intercept was calculated to be close to 0 (-0.03 to -0.06).
Clinical Studies:
Clinical Performance: Evaluated at four institutions in the U.S. using fresh, prospectively collected EDTA whole blood specimens and frozen lysates from patients with CML. Performance was compared to an FDA-cleared molecular assay. A total of 209 specimens were tested. 97.1% (203/209) of results were successful on the first attempt, and 99.5% (208/209) upon retesting. Of 147 quantitative results within the reportable range for both assays, Deming regression analysis showed a slope of 1.0266 and intercept of 0.0600. The predicted bias at MMR (MR3) was MR0.1244 (95% CI: 0.0969 - 0.1519). Bland-Altman difference analysis showed a mean difference (bias) of 0.1336 with a SD of 0.3354. 96.6% (142/147) of results were within the 2SD range.
Precision and Reproducibility Study: Evaluated precision and reproducibility in a multisite study using a panel of eleven samples (negative, near LoD, and at MR levels 1-4 for both e13a2/b2a2 and e14a2/b3a2). Each panel member was tested in duplicate two times per day on four different days by three different operators at three different sites, using three lots of kits. The observed total standard deviation for samples at MR1, MR2, and MR3 was
§ 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).
0
Image /page/0/Picture/0 description: The image contains two logos. The logo on the left is the Department of Health & Human Services logo. The logo on the right is the FDA logo, which stands for U.S. Food & Drug Administration. The FDA logo is in blue.
September 27, 2019
Cepheid Sudhakar Marla, Ph.D. Senior Director, Regulatory Affairs 904 Caribbean Drive Sunnyvale, California 94089
Re: K190076
Trade/Device Name: Xpert BCR-ABL Ultra, GeneXpert Dx System, GeneXpert Infinity-48s and GeneXpert Infinity-80 Systems Regulation Number: 21 CFR 866.6060 Regulation Name: BCR-ABL quantitation test Regulatory Class: Class II Product Code: OYX, OOI Dated: August 15, 2019 Received: August 19, 2019
Dear Sudhakar Marla:
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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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: CDRH does not evaluate information related to contract liability 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
1
gencies. You must comply with all the Act’s
provisions (21 CFR Part 807) labeling (21 CFR Part
Page 2
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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
for Reena Philip, Ph.D. Director Division of Molecular Genetics and Pathology OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K190076
Device Name Xpert® BCR-ABL Ultra
Indications for Use (Describe)
The Xpert BCR-ABL Ultra test is an in vitro diagnostic test for the quantitation of BCR-ABL1 and ABL1 mRNA transcripts in peripheral blood specimens of diagnosed t(9;22) positive Chronic Myeloid Leukemia (CML) patients expressing BCR-ABLI fusion transcripts type e13a2 and/or e14a2. The test utilizes automated, quantitative, real-time reverse transcription polymerase chain reaction (RT-qPCR). The Xpert BCR-ABL Ultra test is intended to measure BCR-ABL1 to ABL1 percent ratios on the International Scale (IS), and also expressed as a log molecular reduction (MR value) from a baseline of 100% (IS), in t(9;22) positive CML patients during of treatment with Tyrosine Kinase Inhibitors (TKIs).
The test does not differentiate between e13a2/62a2 or e14a2/63a2 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 Xpert BCR-ABL Ultra test is intended for use only on the Cepheid GeneXpert® Dx System and the GeneXpert Infinity System.
Type of Use (Select one or both, as applicable)
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) |
|---|
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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3
510(k) Summary
As required by 21 CFR Section 807.92(c).
| Submitted by: | Cepheid
904 Caribbean Drive
Sunnyvale, CA 90489
Phone number: (408) 548-8946 |
|-------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------|
| Contact: | Sudhakar Marla, Ph.D. |
| Date of Preparation: | September 24, 2019 |
| Device: | |
| Trade name: | Xpert® BCR-ABL Ultra |
| Common name: | Xpert BCR-ABL Ultra |
| Type of Test: | Reverse transcription, quantitative, polymerase chain reaction
(RT-qPCR) based nucleic acid amplification |
| Regulation number,
Classification name,
Product code: | 21 CFR 866.6060, BCR-ABL quantitation test, OYX
21 CFR 862.2570, Instrumentation for clinical multiplex test
systems, OOI |
| Classification
Advisory Panel | Pathology (88) |
| Prescription Use | Yes |
| Predicate Device | Asuragen QuantideX qPCR BCR-ABL IS Kit
[DEN160003] |
4
Device Intended Use:
The Xpert BCR-ABL Ultra test is an in vitro diagnostic test for the quantitation of BCR-ABL1 and ABL1 mRNA transcripts in peripheral blood specimens of diagnosed t(9:22) positive Chronic Myeloid Leukemia (CML) patients expressing BCR-ABLI fusion transcripts type e13a2 and/or e14a2. The test utilizes automated, quantitative, real-time reverse transcription polymerase chain reaction (RT-qPCR). The Xpert BCR-ABL Ultra test is intended to measure BCR-ABL1 to ABL1 percent ratios on the International Scale (IS), and also expressed as a log molecular reduction (MR value) from a baseline of 100% (IS), in t(9:22) positive CML patients during monitoring of treatment with Tyrosine Kinase Inhibitors (TKIs).
The test does not differentiate between e13a2/b2a2 or e14a2/b3a2 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 Xpert BCR-ABL Ultra test is intended for use only on the Cepheid GeneXpert® Dx System and GeneXpert Infinity System.
Special conditions for use statement(s):
For in vitro diagnostic use only
For Prescription use only
Device Description:
The Xpert BCR-ABL Ultra test is an automated in vitro diagnostic test for quantifying the amount of BCR-ABLI (BCR-ABL, hereafter) mRNA transcript as a ratio of BCR-ABL/ABL per the International Scale (IS).
The test is performed on the Cepheid GeneXpert® Dx System and GeneXpert Infinity System (referred to as the GeneXpert systems). The GeneXpert systems require the use of single-use, disposable cartridges that hold the PCR reagents and host the PCR process. Because the cartridges are self-contained and specimens never come into contact with working parts of the instrument modules, cross-contamination between samples is minimized. The GeneXpert systems have 1 to 80 randomly accessible modules, depending upon the instrument, that are each capable of performing separate sample preparation and real-time PCR and RT-PCR tests. Each module contains a syringe drive for dispensing fluids (i.e., the syringe drive activates the plunger that works in concert with the rotary valve in the cartridge to move fluids between chambers), and a proprietary I-CORE® thermocycler for performing real-time PCR and RT-PCR and detection.
5
The Xpert BCR-ABL Ultra test includes reagents to detect BCR-ABL fusion genes resulting from two major breakpoints, translocation e13a2/b2a2 and e14a2/b3a2 and the ABL transcript as an endogenous control in peripheral blood specimens. The amount of BCR-ABL transcript in the patient sample is reported as a percent ratio of BCR-ABL/ABL on the International Scale (IS), and also expressed as a log molecular reduction (MR value) from a baseline of 100% (IS), using the GeneXpert software.
There are two controls included in each Xpert BCR-ABL Ultra test, which are the ABL Endogenous Control and the Probe Check Control (PCC). The ABL Endogenous Control normalizes the BCR-ABL target and ensures that sufficient sample is used in the test. The PCC verifies reagent rehydration. PCR tube filling, and that all reaction components, including probes and dyes, are present and functional in the cartridge.
A description of the reagents provided in the kit is described below in Table 1.
| Item | Description | Use |
|---|---|---|
| Proteinase K | Serine protease | Digests proteins and |
| inactivates nucleases in EDTA | ||
| whole blood specimen during | ||
| the sample preparation and | ||
| nucleic acid purification steps | ||
| Lysis Reagent | Guanidinium chloride | |
| buffered solution | Denaturant used to lyse cells, | |
| release of nucleic acids and | ||
| decrease nuclease activity | ||
| during the sample preparation | ||
| step | ||
| Wash Reagent | Guanidinium thiocyanate and | |
| ethanol solution | Reagent used to remove | |
| cellular contaminants during | ||
| nucleic acid binding step | ||
| Xpert BCR-ABL Ultra | ||
| Cartridges | ||
| with Integrated Reaction | ||
| Tubes | Single-use test cartridges that | |
| house, buffered solutions | ||
| (rinse and elution), | ||
| lyophilized beads containing | ||
| reverse transcriptase, DNA | ||
| polymerase, primers and | ||
| probes | Reagents used to perform the | |
| on-board nucleic acid | ||
| isolation, purification and real- | ||
| time RT-qPCR. | ||
| CD/Software | Compact disc | Contains assay definition file |
| (ADF), instruction to import | ||
| ADF into GeneXpert software | ||
| and information for use |
Table 1. Reagents in the Xpert BCR-ABL Ultra Test Kit
6
The peripheral blood specimens are collected in EDTA blood tubes following the user institution's standard procedures and can be stored for up to 72 hours prior to use when stored at 4℃. The test sample, from a peripheral blood specimen, is prepared in a sample preparation tube provided with the kit, with reagents that are provided with the kit (proteinase K and lysis reagent) plus user-supplied ethanol, following instructions in the package insert. After sample preparation, the prepared sample and one reagent provided with the kit (Wash Reagent) are pipetted to separate chambers of the Xpert BCR-ABL Ultra cartridge. The user initiates a test from the system user interface and places the cartridge into the GeneXpert instrument platform, which performs hands-off additional sample preparation and nested, real-time, quantitative reverse transcription polymerase chain reaction (RT-qPCR) for detection of RNA. In this platform, the additional sample preparation, amplification, and real-time detection are all fully-automated and completely integrated. The time to result for the Xpert BCR-ABL Ultra test including offline sample preparation and real-time RT-qPCR is approximately 2.25 hours.
Substantial Equivalence:
The Xpert BCR-ABL Ultra is substantially equivalent to the Asuragen QuantideX qPCR BCR-ABL IS Kit [DEN160003]. The performance of the Xpert BCR-ABL Ultra test was evaluated in a multi-site clinical study using whole blood specimens and was compared to a FDA-cleared molecular assay. The results from the clinical study demonstrated that the performance of Xpert BCR-ABL Ultra is substantially equivalent to the predicate device.
Table 2 shows the similarities and differences between the Xpert BCR-ABL Ultra test and the predicate device.
7
| Similarities | ||
|---|---|---|
| Item | Device | Predicate |
| Intended Use | For the quantitation of BCR- | |
| ABL1 and ABL mRNA | ||
| transcripts in peripheral blood | ||
| specimens of diagnosed t(9;22) | ||
| positive Chronic Myeloid | ||
| Leukemia (CML) patients | ||
| expressing BCR ABL1 fusion | ||
| transcripts type e13a2 and/or | ||
| e14a2. | Same | |
| Target | ||
| Population/Indication | CMP-positive patients during | |
| monitoring of treatment with | ||
| Tyrosine Kinase Inhibitors | ||
| (TKIs) | Same | |
| Test Limitation | The test does not differentiate | |
| between e13a2/b2a2 or | ||
| e14a2/b3a2 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. | Same | |
| Measurand | BCR-ABL1 fusion transcripts | |
| (e13a2/b2a2 and/or e14a2/b3a2) | ||
| and the ABL1 endogenous | ||
| control mRNA | Same | |
| Measurement type | Quantitative | Same |
| Principle of Assay | Reverse transcription, | |
| quantitative, polymerase chain | Same | |
| Similarities | ||
| Item | Device | Predicate |
| reaction (RT-qPCR) based | ||
| nucleic acid amplification | ||
| Traceability Standard | 1st World Health Organization | |
| (WHO) International Genetic | ||
| Reference Panel for quantitation | ||
| of BCR-ABL translocation by | ||
| RQ-PCR | Same | |
| Units | Both % (IS) and Molecular | |
| Response (MR) | Same | |
| Specimen Type | Whole Blood (EDTA) | Same |
Table 2: Comparison of Similarities and Differences of Xpert BCR-ABL Ultra with the Predicate Device
8
| Differences | |||
|---|---|---|---|
| Item | Device | Predicate | |
| Extraction and Assay | |||
| Preparation | Single Use cartridge | Extraction steps may be | |
| manual followed by | |||
| manual assay | |||
| preparation | |||
| Instrument | Cepheid GeneXpert® | ||
| Dx | |||
| GeneXpert Infinity-48s, | |||
| and GeneXpert Infinity- | |||
| 80 | Applied Biosystems | ||
| 7500 Fast Dx Real Time | |||
| PCR Instrument | |||
| Input Range | RNA is isolated from | ||
| white blood cell count | RNA input range of 1 to | ||
| 5µg |
9
| Differences | ||
|---|---|---|
| Item | Device | Predicate |
| (WBCC) range | ||
| $1.5 x 10^5$ to $3.0 x 10^7$ | ||
| cells/mL | ||
| Controls | Probe Check Control | Three controls RNA |
| High (MR 1.5), mRNA | ||
| Low (MR 3.5), and RNA | ||
| Negative. Each provided | ||
| in separate tubes | ||
| Calibrators | None provided. Each lot | |
| of Xpert BCR-ABL | ||
| Ultra is calibrated to | ||
| secondary standards that | ||
| were calibrated to the | ||
| World Health | ||
| Organization (WHO) | ||
| international genetic | ||
| reference panel for | ||
| quantitation of BCR- | ||
| ABL transcript. | Four levels formulated | |
| to MR1.0, 2.0, 3.0, 4.0 | ||
| traceable to the World | ||
| Health Organization | ||
| (WHO) international | ||
| genetic reference panel | ||
| for quantitation of | ||
| BCR-ABL transcript. |
The Xpert BCR-ABL Ultra test has the same general intended use as the predicate device and has the same technological characteristics as the predicate device. The differences between the Xpert BCR-ABL Ultra test and the predicate device do not raise different questions of safety and effectiveness. The clinical study demonstrates that the Xpert BCR-ABL Ultra test is acceptable for its intended use and is substantially equivalent to the predicate device described above.
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Non-Clinical Studies:
Linearity/Dynamic Range
Linearity was evaluated independently for each of the two major breakpoints, e13a2/b2a2 and e14a2/b3a2, using CML clinical specimens that were specific for a high level of either the e13a2/b2a2 or e14a2/b3a2 breakpoint. Lysate from each high level of BCR-ABL transcript CML specimen was diluted in a background lysate prepared from CML negative clinical specimen to target ranges of ~50% (IS)/MR0.30 to 0.000625% (IS)/MR5.20. The panel members, including the negative level, were tested on two assay kit lots in replicates of 4 per kit lot.
Testing and statistical analyses were conducted in accordance with CLSI EP06-A. Linear regression analyses were performed for first, second and third order polynomials. The results for each breakpoint were considered linear if the polynomial regression coefficients were insignificant (p-values > 0.05). The linear regression curves for both transcripts are shown in Figure 1 and Figure 2 below.
Image /page/10/Figure/4 description: The figure shows linear regression curves for breakpoint transcript e13a2/b2a2. The x-axis represents the expected MR, while the y-axis represents the output MR and output BCR-ABL % (IS). The plot includes data points and three regression models: simple linear regression, second-order polynomial, and third-order polynomial. Equations for each model are displayed, such as y = -0.05833 + 0.99501x with R^2 = 0.98304 for the simple linear regression model.
Figure 1: Linear Regression Curves for Breakpoint Transcript
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Image /page/11/Figure/0 description: The image is a scatter plot comparing "Output_MR" to "e14a2/b3a2 Expected_MR". The plot includes data points labeled as "Output MR" and three trendlines: "Simple Linear Regression Model", "Second-Order Polynomial Model", and "Third-Order Polynomial Model". Equations for each trendline are displayed, such as "y = 0.03647 + 1.03153x, R2 = 0.97876" for the linear regression model.
Figure 2: Linear Regression Curves for Breakpoint Transcript e14a2/b3a2
The estimated regression intercepts, slopes and R2 values from the linear model are shown in Table 3.
| Breakpoint | Intercept | Slope | R2 |
|---|---|---|---|
| e13a2/b2a2 | -0.05833 | 0.99501 | 0.98304 |
| e14a2/b3a2 | 0.03647 | 1.03153 | 0.9788 |
Table 3: Regression Coefficients from Linear Model
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Collectively, the data support an observation of linearity from at least 55% (IS)/MR 0.26 to ~0.0019% (IS)/MR4.75 with a maximum SD of 0.26. The reportable range spans from the limits of linearity at 55% (IS)/MR0.26 to the LoD/LOQ at 0.0030% (IS)/MR4.52.
Analytical Sensitivity
The limit of detection (LoD) was estimated for both e13a2/b2a2 and e14a2/b3a2 breakpoints by testing serial dilutions of High CML positive specimens [>10% (IS)/MR1] as well as testing Low CML positive specimens [MR4.52/1% (IS)/MR3). Test controls consisted of CML clinical specimens in EDTA whole blood at the respective BCR-ABL transcript level without the interfering substance. Each CML specimen was tested in the absence and presence of the five individual interferents at 4 replicates per condition.
A substance was considered non-interfering if in its presence the % mean (IS)/MR ratio observed was within 3-fold difference when compared to the control.
No clinically significant inhibitory effects on the Xpert BCR-ABL Ultra test were observed with any of the interfering substances evaluated in this study. Although some variability and statistically significant differences (p-value