(82 days)
The QUANTA Flash® ß2GP1-Domain1 is an in vitro chemiluminescent immunoassay (CIA) for the semi-quantitative determination of IgG autoantibodies to B2GP1-Domain1 in human serum or citrated plasma. The presence of anti-S2GP1-Domain1 autoantibodies is used in conjunction with clinical and other laboratory findings as an aid in the diagnosis of antiphospholipid syndrome. The QUANTA Flash® ß2GP1-Domain1 is not intended to replace assays for antibodies against the whole B2GP1 molecule. Testing for antibodies to the whole is required according to the classification criteria for antiphospholipid syndrome.
The QUANTA Flash® ß2GP1-Domain1 Controls are intended for quality control purposes of the QUANTA Flash® ß2GP1-Domain1 chemiluminescent immunoassay (CIA) kit.
The HemosIL® AcuStar Anti-S2GPI Domain 1 is an in vitro chemiluminescent immunoassay (CIA) for the semiquantitative determination of IgG autoantibodies to 82GPI Domain 1 in human serum or citrated plasma. The presence of anti-ß2GPI Domain 1 autoantibodies is used in conjunction with clinical and other laboratory findings as an aid in the diagnosis of antiphospholipid syndrome. The HemosIL® AcuStar Anti-S2GPI Domain 1 is not intended to replace assays for antibodies against the whole 82GPI molecule. Testing for antibodies to the whole 13GPI molecule is required according to the classification criteria for antiphospholipid syndrome.
The HemosIL AcuStar Anti-B2GPI Domain 1 Controls are intended for quality control purposes of the HemosIL AcuStar Anti-ß2GPI Domain 1 chemiluminescent immunoassay (CIA) kit.
The QUANTA Flash® ß2GP1-Domain1 assay is designed to run on the BIO-FLASH® instrument. This platform is a fully automated closed system with continuous load and random access capabilities that automatically processes the samples, runs the assay and reports the results. It includes liquid handling hardware, luminometer and computer with software-user interface. The QUANTA Flash® ß2GP1-Domain1 assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH instrument.
The assays included in this submission, the QUANTA Flash ®2GP1-Domain1 marketed by Inova Diagnostics Inc. (9900 Old Grove Road, San Diego, CA 92131) and HemoslL "AcuStar Anti-ß2GPI Domain 1 marketed by Instrumentation Laboratory (180 Hartwell Road Bedford, MA 01730), are equivalent assays. Therefore all data stated hereafter and referred to as: QUANTA Flash ß2GP1-Domain1 data is equivalently also valid for HemosIL * AcuStar Anti-ß2GPI Domain 1.
Recombinant ß2GP1-Domain1 protein is coated onto paramagnetic beads, which are stored lyophilized in the reagent cartridge. The reagent pack is prepared for use in the BIO-FLASH system by pressing down on the grey lid of the reagent pack to pierce the induction seals on the reagent tubes. Once the seals are broken, the beads are rehydrated by adding the entire contents of the vial of resuspension buffer to the bead reagent tube using the transfer pipette supplied with the kit. Only the hole above the bead reagent tube is accessible at this point. The beads are then mixed with the resuspension buffer by pipetting up and down 30 times. This amount of mixing ensures complete resuspension of the beads. The label covering the remaining three reagent holes is now removed, and the reagent cartridge is then loaded onto the BIO-FLASH instrument. Samples are also loaded onto the instrument in sample racks. A patient serum sample is prediluted 1:10 by the BIO-FLASH with system rinse in a small disposable plastic cuvette. Small amounts of the diluted patient serum, the beads, and assay buffer are all combined into a second cuvette, and mixed. This cuvette is then incubated at 37°C. The beads are magnetized and washed several times. lsoluminol conjugated anti-human IgG antibodies are then added to the cuvette, and again incubated at 37°C. The beads are magnetized and washed repeatedly. The isoluminol conjugate is oxidized when Trigger 1 (Fe(III) coproporphyrin in sodium hydroxide solution) and Trigger 2 (urea-hydrogen peroxide in sodium chloride solution) are added to the cuvette, and the flash of light produced from this reaction is measured as Relative Light Units (RLU) by the BIO-FLASH optical system. The RLU are proportional to the amount of isoluminol conjugate that is bound to the human IgG, which is in turn proportional to the amount of anti- ß2GP1-Domain1 antibodies bound to the corresponding ß2GP1-Domain1 on the beads.
The QUANTA Flash® ß2GP1-Domain1 assay utilizes a predefined lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. Every new lot number of reagent cartridge must be calibrated before first use, with the QUANTA Flash® ß2GP1-Domain1 Calibrators. Based on the results obtained with the two Calibrators included in the reagent kit, an instrument specific Working Curve is created, which is used to calculate chemiluminescent units (CU) from the instrument signal (RLU) obtained for each sample.
The QUANTA Flash® ß2GP1-Domain1 kit contains the following materials:
One (1) QUANTA Flash ß2GP1-Domain1 Reagent Cartridge, containing the following reagents for 50 determinations:
- ß2GP1-Domain1 antigen coated paramagnetic beads in a suspension. a.
- b. Assay Buffer – buffer containing protein stabilizers and preservatives.
- C. Tracer IgG - Isoluminol labeled anti-human IgG antibodies in buffer, containing protein stabilizers and preservative.
- d. Sample Diluent - buffer containing protein stabilizers and preservatives.
- d. Resuspension Buffer - buffer containing protein stabilizers and preservatives.
- e. QUANTA Flash ß2GP1-Domain1 Calibrator 1: One (1) barcode labeled tube containing 1.0 mL prediluted, ready to use reagent. Calibrators contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives.
- f. QUANTA Flash ß2GP1-Domain1 Calibrator 2: One (1) barcode labeled tube containing 1.0 mL prediluted, ready to use reagent. Calibrators contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives
The QUANTA Flash® ß2GP1-Domain1 Controls kit contains three vials of Low Control and three vials of High Control.
- QUANTA Flash ß2GP1-Domain1 Low Control: Three (3) barcode labeled tubes containing 1.0 mL, ready to use reagent. Controls contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives.
- QUANTA Flash ß2GP1-Domain1 High Control: Three (3) barcode labeled tubes containing 1.0 mL, ready to use reagent. Controls contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives.
Here's a breakdown of the acceptance criteria and study details for the QUANTA Flash® ß2GP1-Domain1 device, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
| Parameter | Acceptance Criteria | Reported Device Performance |
|---|---|---|
| Sample Matrix Comparison | ||
| Weighted R | $\geq 0.975$ | 0.997 |
| Intercept (Constant bias) | $\pm 3 CU$ | 0.1 |
| Slope (Proportional bias) | 0.9 - 1.1 | 1.01 |
| Weighted S y/x | $\leq 0.5$ | 0.064 |
| Predicted bias at cut-off | $\pm 3 CU$ | 0.3 |
| 95% Cl of the bias | <20% $\pm 4 CU$ | 0.00 to 1.03 |
| Precision (Serum) | Total %CV: < 15% | 5.6-10.6% (for 8 samples with varying concentrations) |
| Precision (Plasma) | Total %CV: < 15% | 2.5-7.8% (for 3 samples with varying concentrations) |
| Linearity (Recovery) | 80-120%, or $\pm 4 CU$, whichever is greater | Serum: 83.9%-113.9% (combined for 5 samples) Plasma: 86.6%-111.9% (combined for 3 samples) |
| Linearity (Linear Regression) | Slope: 0.9-1.1, R$^2$: $\geq 0.95$ | Serum: Slope 1.01, R$^2$ 1.00 (Combined for 5 samples) Plasma: Slope 0.96, R$^2$ 0.99 (Combined for 3 samples) |
| Interference | 85%-115% recovery, or $\pm 4$ CU difference, whichever is greater, compared to control samples. | Bilirubin (up to 100 mg/dL): 92.9% to 109.4% Hemoglobin (up to 200 mg/dL): 95.6% to 106.3% or 1.7 CU Triglycerides (up to 1000 mg/dL): 92.5% to 107.7% Cholesterol (up to 224.3 mg/dL): 92.5% to 107.7% RF IgM (up to 500 IU/mL): 86.2% to 109.9% or 0.9 CU |
| Lot to Lot Comparison | ||
| Weighted R | $\geq 0.975$ | RP3 vs RP11: 0.996 RP11 vs PP1: 0.998 PP1 vs RP3: 0.986 |
| Intercept (constant bias) | $\pm 3$ CU | RP3 vs RP11: 1.9 RP11 vs PP1: 0.06 PP1 vs RP3: -1.3 |
| Slope (Proportional bias) | 0.9 - 1.1 | RP3 vs RP11: 1.03 RP11 vs PP1: 1.04 PP1 vs RP3: 1.00 |
| Weighted S y/x | $\leq 0.5$ | RP3 vs RP11: 0.07 RP11 vs PP1: 0.05 PP1 vs RP3: 0.13 |
| Predicted bias at cut-off | $\pm 3$ CU | RP3 vs RP11: 2.5 RP11 vs PP1: 0.8 PP1 vs RP3: -1.2 |
| 95% CI of the bias | $\leq 20%$ or $\pm 4$ CU | RP3 vs RP11: 1.1 to 3.8 RP11 vs PP1: -0.3 to 1.8 PP1 vs RP3: -3.9 to 1.6 |
| Shelf-Life (Accelerated Stability - Beads, Resuspension Buffer, Reagent Kit) | Lower and upper 95% Cl interval of regression line between 85% and 115% recovery at day 14; no individual data point $\leq 75%$ or $\geq 125%$ recovery at day 14. | All components fulfilled acceptance criteria. |
| Shelf-Life (Accelerated Stability - Controls and Calibrators) | Lower and upper 95% Cl interval of regression line between 90% and 110% recovery at day 14; no individual data point $\leq 80%$ or $\geq 120%$ recovery at day 14. | All components fulfilled acceptance criteria. |
| Calibrators Onboard Stability | All 5 calibrations successful in 8.5 hours; average Calibrator RLU recovery 90-110% compared to first use; all Controls and patient panel in expected range. | 5 successful calibrations in 8.5 hours; Calibrator RLU values within 90-110%; all Controls and patient panel samples ran within expected range. Supports 4 calibrations over 8 hours. |
| Controls Onboard Stability | All values within established range for all runs; linear regression line for %recovery between 85% and 115% at run 15. | All controls ran within acceptable ranges for all runs (20 runs); regression line remained between 85% and 115% at run 15. Supports 15 uses, 10 min/use. |
| Reagent Cartridge Onboard Stability | Stability claim at actual measurement day preceding 95% CI of regression line reaching 85% or 115% recovery, OR preceding 2 data points or $\geq 2%$ of recovery data $\leq 75%$ or $\geq 125%$ recovery. | None of the endpoints reached within the test period. Onboard stability set at 60 days. |
| Real-Time Stability (Reagent Cartridge, Calibrators, Controls) | Results within respective QC ranges for QC panel samples (reagent cartridge, calibrators); individual values within acceptable ranges compared to release values (controls). | All results to date (up to 12-15 months) were within acceptance limits. One year expiration dating verified. |
| Sample Storage (Room Temp) | Recovery between 85-115% for positive samples; 80-120% or 4 CU for negative samples. | All results fulfilled criteria up to 48 hours. |
| Sample Storage (2-8°C) | Recovery between 85-115% for positive samples; 80-120% or 4 CU for negative samples. | All results fulfilled criteria up to 14 days. |
| Sample Storage (Freeze/Thaw) | Recovery between 85-115% for positive samples; 80-120% or 4 CU for negative samples. | All results fulfilled criteria up to 3 freeze/thaw cycles. |
| Clinical Sensitivity (APS) | Not explicitly stated as a numerical acceptance criterion, but result is provided. | 51.1% (95% CI: 45.0-57.2%) |
| Clinical Specificity (Controls) | Not explicitly stated as a numerical acceptance criterion, but result is provided. | 99.6% (95% CI: 98.9-99.9%) |
| ROC AUC | Not explicitly stated as a numerical acceptance criterion, but result is provided. | 0.84 (95% CI: 0.81 to 0.86) |
| Expected values (Normal Population) | Not explicitly stated as a numerical acceptance criterion, but result is provided. | 0.25% positive (1/400 samples) in healthy blood donors. |
| Cross-Reactivity | Negligible positivity rate in specified autoimmune diseases (Crohn's Disease, Ulcerative Colitis, Rheumatoid Arthritis, Osteoarthritis, Scleroderma) and infectious diseases (Hepatitis B, Hepatitis C, Syphilis). | Total controls (n=613): 0.5% positive (3 samples). Specifically: Crohn's Disease: 1.0% (1/104); Scleroderma: 1.6% (2/127); all others 0%. No cross-reactivity detected. |
(Note: "N/A" is used if the information is not explicitly stated in the provided text.)
2. Sample Size Used for the Test Set and Data Provenance
- Sample Matrix Comparison: 79 matched serum and citrated plasma samples. Data provenance is not specified (e.g., country of origin, retrospective/prospective).
- Precision (Serum): 8 serum samples. Data provenance not specified.
- Precision (Plasma): 3 plasma samples. Data provenance not specified.
- Limit of Blank (LoB) and Limit of Detection (LoD): 140 determinations (60 blank samples, 80 low-level samples). Data provenance not specified.
- Auto-rerun function: 3 high positive specimens. Data provenance not specified.
- High concentration hook effect: 2 high positive samples. Data provenance not specified.
- Linearity (Serum): 5 serum samples. Data provenance not specified.
- Linearity (Plasma): 3 plasma samples. Data provenance not specified.
- Interference: 5 specimens (3 serum, 2 plasma). Data provenance not specified.
- Cross-reactivity: 613 control samples from patients with autoimmune diseases or infections. Data provenance not specified.
- Lot to Lot Comparison: 26 samples. Data provenance not specified.
- Shelf-Life (Accelerated Stability): Various sealed components (Reagent Kit, beads, Resuspension Buffer, Calibrators, Controls) – quantities not specified beyond number of lots (e.g. 1 lot Reagent Kit, 3 lots beads).
- Calibrators Onboard Stability: Calibrators (number not specified); Controls and a panel of characterized patient specimens (number not specified).
- Controls Onboard Stability: 2 vials of each Control; patient panel samples (number not specified).
- Reagent Cartridge Onboard Stability: Three lots of reagent cartridge; up to 4 serum specimens; Low and High Controls.
- Real-Time Stability: QC panel samples (number not specified); Calibrators, Controls (number not specified beyond "all results to date").
- Sample Stability: 8 samples (4 serum, 4 plasma). Data provenance not specified.
- Cut-off, Reference Range: 30 subjects (5 healthy blood donors, 4 viral hepatitis, 10 SLE without thrombotic events, 10 Syphilis, 1 HIV). All serum matrix. Data provenance not specified.
- Clinical Sensitivity, Specificity: 1090 characterized samples (270 APS, 71 infectious diseases, 566 other diseases, 183 conditions "without APS"). These samples were not used for establishing the reference range. Data provenance is not specified beyond "characterized samples."
- Expected Values (Normal Population): 400 apparently healthy blood donors. Data provenance not specified.
- Comparison with Predicate Device: 238 samples (all samples from clinical validation study within reportable range + 8 additional contrived samples). Data provenance not specified.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The text does not specify the number of experts or their qualifications used to establish the ground truth for the test set. It mentions "characterized samples" for the clinical validation study and "clinical and other laboratory findings" as an aid in diagnosis, but no details on expert review or adjudication.
4. Adjudication Method for the Test Set
The text does not specify any adjudication method (e.g., 2+1, 3+1, none) for the test set.
5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
This device is a chemiluminescent immunoassay (CIA) for the semi-quantitative determination of autoantibodies. It is an in-vitro diagnostic (IVD) device that performs automated laboratory testing. Therefore, an MRMC study related to "human readers improving with AI vs without AI assistance" is not applicable to this type of device. The device itself is the "reader" or analytical instrument.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the studies described are for the device (the immunoassay kit and BIO-FLASH instrument system) operating in a standalone manner, generating semi-quantitative results without human intervention in the measurement process. Human intervention would be for sample collection, preparation, loading, and interpretation of the results by a clinician. The performance metrics (sensitivity, specificity, precision, linearity, etc.) are for the algorithm and assay system as a whole.
7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.)
The ground truth for the clinical studies (clinical sensitivity and specificity) is based on diagnosis of Antiphospholipid Syndrome (APS), as well as diagnoses of infectious diseases, other autoimmune diseases (Crohn's, UC, RA, Osteoarthritis, Scleroderma), and conditions "without APS" (pre-eclampsia/eclampsia, fetal loss no APS, SLE no APS, thrombosis no APS, atopic dermatitis). The specific methods for establishing these clinical diagnoses (e.g., based on established clinical criteria, expert consensus, pathology, or long-term outcomes) are not detailed in the provided text.
For analytical studies, the "ground truth" often refers to reference methods or known concentrations, which are implied for linearity, precision, and interference studies (e.g., spiking known concentrations).
8. The Sample Size for the Training Set
The text does not specify a separate "training set" sample size in the context of machine learning or AI models. This device is an immunoassay, the "training" equivalent would be the development and optimization of the assay reagents, protocols, and the establishment of master curves.
9. How the Ground Truth for the Training Set Was Established
Similarly, as there's no explicitly mentioned "training set" in an AI/ML context, the concept of "ground truth for the training set" isn't directly applicable. The assay's performance and cut-offs were established through various analytical and clinical studies described (e.g., cutoff established using 99th percentile of 30 subjects, analytical measuring range defined by master curve standards), which could be seen as the data used to "train" or validate the assay's operational parameters.
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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
February 4, 2016
INOVA DIAGNOSTICS, INC. DR. GABRIELLA LAKOS DIRECTOR OF RESEARCH AND DEVELOPMENT 9900 OLD GROVE ROAD SAN DIEGO, CA, 92131
Re: K152875 Trade/Device Name: QUANTA Flash® ß2GP1-Domain1 OUANTA Flash® ß2GP1-Domain1 Controls HemosIL® Acustar Anti-ß2GPI-Domain 1 HemosIL® Acustar Anti-ß2GPI-Domain 1 Controls Regulation Number: 21 CFR § 866.5660 Regulation Name: Multiple autoantibodies immunological test system Regulatory Class: II Product Code: MSV, JJX Dated: September 29, 2015 Received: October 5, 2015
Dear Dr. Lakos:
This letter corrects our substantially equivalent letter of December 21, 2015.
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: 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 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 (reporting of
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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.
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809). please contact the Division of Industry and Consumer Education 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" (21 CFR 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/ReportaProblem/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 Industry and Consumer Education 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.
Sincerely yours.
Kelly Oliner -S
FOR
Leonthena R. Carrington, MS, MBA, MT(ASCP) Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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510(k) Number (if known) K152875
Device Name QUANTA Flash® ß2GP1-Domain1
Indications for Use (Describe)
The QUANTA Flash® ß2GP1-Domain1 is an in vitro chemiluminescent immunoassay (CIA) for the semi-quantitative determination of IgG autoantibodies to B2GP1-Domain1 in human serum or citrated plasma. The presence of anti-S2GP1-Domain1 autoantibodies is used in conjunction with clinical and other laboratory findings as an aid in the diagnosis of antiphospholipid syndrome. The QUANTA Flash® ß2GP1-Domain1 is not intended to replace assays for antibodies against the whole B2GP1 molecule. Testing for antibodies to the whole is required according to the classification criteria for antiphospholipid syndrome.
| 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) |
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov
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510(k) Number (if known) K152875
Device Name QUANTA Flash® ß2GP1-Domain1 Controls
Indications for Use (Describe)
The QUANTA Flash ß2GP1-Domain1 Controls are intended for quality control purposes of the QUANTA Flash ß2GP1-Domain1 chemiluminescent immunoassay (CIA) kit.
| 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) |
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov
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510(k) Number (if known) K152875
Device Name HemosIL® AcuStar Anti-ß2GPI Domain 1
Indications for Use (Describe)
The HemosIL® AcuStar Anti-S2GPI Domain 1 is an in vitro chemiluminescent immunoassay (CIA) for the semiquantitative determination of IgG autoantibodies to 82GPI Domain 1 in human serum or citrated plasma. The presence of anti-ß2GPI Domain 1 autoantibodies is used in conjunction with clinical and other laboratory findings as an aid in the diagnosis of antiphospholipid syndrome. The HemosIL® AcuStar Anti-S2GPI Domain 1 is not intended to replace assays for antibodies against the whole 82GPI molecule. Testing for antibodies to the whole 13GPI molecule is required according to the classification criteria for antiphospholipid syndrome.
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) |
|---|---|
| ------------------------------------------------------------------------------------- | ----------------------------------------------- |
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov
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510(k) Number (if known) K152875
Device Name
HemosIL® AcuStar Anti-ß2GPI Domain 1 Controls
Indications for Use (Describe)
The HemosIL AcuStar Anti-B2GPI Domain 1 Controls are intended for quality control purposes of the HemosIL AcuStar Anti-ß2GPI Domain 1 chemiluminescent immunoassay (CIA) kit.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
| Over-The-Counter Use (21 CFR 801 Subpart C)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov
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510(k) Summary
| QUANTA Flash® β2GP1-Domain1 |
|---|
| QUANTA Flash® β2GP1-Domain1 Controls |
| HemosIL® AcuStar Anti- β₂GPI Domain 1 |
| HemosIL® AcuStar Anti- β₂GPI Domain 1 Controls |
| Page 1 of 19 |
Table of Contents
| Administrative data…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… | |
|---|---|
| Predicate device…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… | |
| Device description | |
| Intended use(s) | |
| Indications for use…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… | |
| Substantial equivalence | |
| Comparison to predicate device | |
| Analytical performance characteristics | |
| Value assignment and traceability of Calibrators and Controls | |
| Sample Matrix Comparison | |
| Precision | |
| Plasma Precision Verification | |
| Limit of Blank (LoB) and Limit of Detection (LoD) | |
| Analytical Measuring Range (AMR) | |
| Auto-rerun function and reportable results | |
| High concentration hook effect | |
| Linearity | |
| Interference | |
| Cross-reactivity | |
| Lot to lot comparison | |
| Stability | |
| Cut-off, reference range | |
| Clinical performance characteristics | |
| Clinical sensitivity, specificity | |
| Expected values | |
| Comparison with predicate device |
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This summary of the 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
Administrative data
| Submitter: | Inova Diagnostics, Inc9900 Old Grove Road,San Diego, CA, 92131 | |
|---|---|---|
| Purpose of submission: | New device(s) | |
| Devices in the submission: | QUANTA Flash® β2GP1-Domain1QUANTA Flash® β2GP1-Domain1 ControlsHemosIL® AcuStar Anti-β2GPI Domain 1HemosIL® AcuStar Anti-β2GPI Domain 1 Controls | |
| Scientific contact: | Gabriella Lakos, Director of Research, RheumatologyInova Diagnostics, Inc.9900 Old Grove Road, San Diego, CA, 92131Phone: 858-586-9900/1393Fax: 858-863-0025email: glakos@inovadx.com | |
| Quality Systems contact: | Ronda Elliott, VP, Quality Systems and RAInova Diagnostics, Inc9900 Old Grove Road, San Diego, CA, 92131Phone: 858-586-9900/1381Fax: 858-863-0025email: relliot@inovadx.com | |
| Device name (assay kit): | Proprietary name: | QUANTA Flash® β2GP1-Domain1HemosIL® AcuStar Anti-β₂GPI Domain 1 |
| Common name: | Anti-β2GPI Domain 1 Chemiluminescent immunoassay | |
| Classification name: | Anti-β2GPI Domain 1 antibody, antigen and control | |
| Regulation Description | Multiple autoantibodies immunological test system | |
| Regulation Medical Specialty | Immunology | |
| Review Panel | Immunology |
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510(k) Summary QUANTA Flash® ß2GP1-Domain1
| Product Code | System, Test, Antibodies,B2-Glycoprotein I (B2-GPI) (MSV) | |
|---|---|---|
| Regulation Number | 866.5660 | |
| Device Class | 2 | |
| Device name (Controls): | Proprietary name: | QUANTA Flash® β2GP1-Domain1 ControlsHemosIL® AcuStar Anti-β2GPI Domain 1 Controls |
| Common name: | Domain1 Controls | |
| Classification name: | single (specified) analyte controls (assayed andunassayed) | |
| Regulation Description | Quality control material (assayed and unassayed) | |
| Regulation Medical Specialty | Clinical Chemistry | |
| Product Code | JJX | |
| Regulation Number | 862.1660 | |
| Device Class | 1 (reserved) |
Predicate device
QUANTA Lite® ß2GPI IgG ELISA, 510(k) number: K970551
Device description
The QUANTA Flash® ß2GP1-Domain1 assay is designed to run on the BIO-FLASH® instrument. This platform is a fully automated closed system with continuous load and random access capabilities that automatically processes the samples, runs the assay and reports the results. It includes liquid handling hardware, luminometer and computer with software-user interface. The QUANTA Flash® ß2GP1-Domain1 assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH instrument.
The assays included in this submission, the QUANTA Flash ®2GP1-Domain1 marketed by Inova Diagnostics Inc. (9900 Old Grove Road, San Diego, CA 92131) and HemoslL "AcuStar Anti-ß2GPI Domain 1 marketed by Instrumentation Laboratory (180 Hartwell Road Bedford, MA 01730), are equivalent assays. Therefore all data stated hereafter and referred to as: QUANTA Flash ß2GP1-Domain1 data is equivalently also valid for HemosIL * AcuStar Anti-ß2GPI Domain 1.
Recombinant ß2GP1-Domain1 protein is coated onto paramagnetic beads, which are stored lyophilized in the reagent cartridge. The reagent pack is prepared for use in the BIO-FLASH system by pressing down on the grey lid of the reagent pack to pierce the induction seals on the reagent tubes. Once the seals are broken, the beads are rehydrated by adding the entire contents of the vial of resuspension buffer to the bead reagent tube using the transfer pipette supplied with the kit. Only the hole above the bead reagent
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tube is accessible at this point. The beads are then mixed with the resuspension buffer by pipetting up and down 30 times. This amount of mixing ensures complete resuspension of the beads. The label covering the remaining three reagent holes is now removed, and the reagent cartridge is then loaded onto the BIO-FLASH instrument. Samples are also loaded onto the instrument in sample racks. A patient serum sample is prediluted 1:10 by the BIO-FLASH with system rinse in a small disposable plastic cuvette. Small amounts of the diluted patient serum, the beads, and assay buffer are all combined into a second cuvette, and mixed. This cuvette is then incubated at 37°C. The beads are magnetized and washed several times. lsoluminol conjugated anti-human IgG antibodies are then added to the cuvette, and again incubated at 37°C. The beads are magnetized and washed repeatedly. The isoluminol conjugate is oxidized when Trigger 1 (Fe(III) coproporphyrin in sodium hydroxide solution) and Trigger 2 (urea-hydrogen peroxide in sodium chloride solution) are added to the cuvette, and the flash of light produced from this reaction is measured as Relative Light Units (RLU) by the BIO-FLASH optical system. The RLU are proportional to the amount of isoluminol conjugate that is bound to the human IgG, which is in turn proportional to the amount of anti- ß2GP1-Domain1 antibodies bound to the corresponding ß2GP1-Domain1 on the beads.
The QUANTA Flash® ß2GP1-Domain1 assay utilizes a predefined lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. Every new lot number of reagent cartridge must be calibrated before first use, with the QUANTA Flash® ß2GP1-Domain1 Calibrators. Based on the results obtained with the two Calibrators included in the reagent kit, an instrument specific Working Curve is created, which is used to calculate chemiluminescent units (CU) from the instrument signal (RLU) obtained for each sample.
The QUANTA Flash® ß2GP1-Domain1 kit contains the following materials:
One (1) QUANTA Flash ß2GP1-Domain1 Reagent Cartridge, containing the following reagents for 50 determinations:
- ß2GP1-Domain1 antigen coated paramagnetic beads in a suspension. a.
- b. Assay Buffer – buffer containing protein stabilizers and preservatives.
- C. Tracer IgG - Isoluminol labeled anti-human IgG antibodies in buffer, containing protein stabilizers and preservative.
- d. Sample Diluent - buffer containing protein stabilizers and preservatives.
- d. Resuspension Buffer - buffer containing protein stabilizers and preservatives.
- e. QUANTA Flash ß2GP1-Domain1 Calibrator 1: One (1) barcode labeled tube containing 1.0 mL prediluted, ready to use reagent. Calibrators contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives.
- f. QUANTA Flash ß2GP1-Domain1 Calibrator 2: One (1) barcode labeled tube containing 1.0 mL prediluted, ready to use reagent. Calibrators contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives
The QUANTA Flash® ß2GP1-Domain1 Controls kit contains three vials of Low Control and three vials of High Control.
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- QUANTA Flash ß2GP1-Domain1 Low Control: Three (3) barcode labeled tubes containing 1.0 mL, ready to use reagent. Controls contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives.
- QUANTA Flash ß2GP1-Domain1 High Control: Three (3) barcode labeled tubes containing 1.0 mL, ready to use reagent. Controls contain human antibodies to ß2GP1-Domain1 in stabilizers and preservatives.
Intended use(s)
QUANTA Flash ß2GP1-Domain1
The QUANTA Flash ß2GP1-Domain1 is an in vitro chemiluminescent immunoassay (CIA) for the semiquantitative determination of IgG autoantibodies to ß2GP1-Domain1 in human serum or citrated plasma. The presence of anti-ß2GP1-Domain1 autoantibodies is used in conjunction with clinical and other laboratory findings as an aid in the diagnosis of antiphospholipid syndrome. The QUANTA Flash 182GP1-Domain1 is not intended to replace assays for antibodies against the whole ß2GP1 molecule. Testing for antibodies to the whole ß2GP1 molecule is required according to the classification criteria for antiphospholipid syndrome.
QUANTA Flash ß2GP1-Domain1 Controls
The QUANTA Flash ß2GP1-Domain1 Controls are intended for quality control purposes of the QUANTA Flash ß2GP1-Domain1 chemiluminescent immunoassay (CIA) kit.
HemosIL AcuStar Anti-ß₂GPI Domain 1
The HemosIL AcuStar Anti-R2GPI Domain 1 is an in vitro chemiluminescent immunoassay (CIA) for the semi-quantitative determination of IgG autoantibodies to ßGPI Domain 1 in human serum or citrated plasma. The presence of anti-ß2GPI Domain 1 autoantibodies is used in conjunction with clinical and other laboratory findings as an aid in the diagnosis of antiphospholipid syndrome. The HemoslL® AcuStar Antiß,GPI Domain 1 is not intended to replace assays for antibodies against the whole ß₂GPI molecule. Testing for antibodies to the whole ß2GPI molecule is required according to the classification criteria for antiphospholipid syndrome.
HemosIL AcuStar Anti-ß2GPI Domain 1 Controls
The HemosIL AcuStar Anti-ß2GPI Domain 1 Controls are intended for quality control purposes of the HemosIL Anti-ß2GPI Domain 1 chemiluminescent immunoassay (CIA) kit.
Indications for use
Same as Intended use.
Substantial equivalence
The QUANTA Flash ß2GP1-Domain1 and the QUANTA Flash ß2GP1-Domain1 Controls have a similar intended use and assay principle as the predicate device.
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Comparison to predicate device
QUANTA Flash® ß2GP1-Domain1 reagent kit
| Similarities | ||
|---|---|---|
| Item | Applicant | Predicate Device |
| QUANTA Flash® ß2GP1-Domain1 | QUANTA Lite® ß₂GPI IgG ELISA | |
| Intended use | The QUANTA Flash® ß2GP1-Domain1 isan in vitro chemiluminescentimmunoassay (CIA) for the semi-quantitative determination of IgGautoantibodies to ß2GP1-Domain1 inhuman serum or citrated plasma. Thepresence of anti-ß2GP1-Domain1autoantibodies is used in conjunctionwith clinical and other laboratoryfindings as an aid in the diagnosis ofantiphospholipid syndrome. TheQUANTA Flash® ß2GP1-Domain1 is notintended to replace assays forantibodies against the whole ß2GP1molecule. Testing for antibodies to thewhole ß2GP1 molecule is requiredaccording to the classification criteriafor antiphospholipid syndrome. | QUANTA Lite® ß2 GPI IgG is an enzymelinked immunoassay (ELISA) for thesemi-quantitative detection of ß2 GPIIgG antibodies in human serum. Thepresence of ß₂ GPI IgG antibodies canbe used in conjunction with clinicalfindings and other laboratory tests toaid in the diagnosis of certainautoimmune disease thromboticdisorders such as those secondary tosystemic lupus erythematosus (SLE) orother lupus-like thrombotic diseases. |
| Assay methodology | Solid phase (heterogeneous)immunoassay | Solid phase (heterogeneous)immunoassay |
| Shelf life | One year | One year |
| Differences | ||
|---|---|---|
| Item | QUANTA Flash β2GP1-Domain1 | Predicate Device |
| Detection/Operating principle | Chemiluminescent immunoassay | Enzyme-linked immunosorbent assay |
| Solid phase | Paramagnetic microparticles (beads) | 96-well plate |
| Antigen | Recombinant domain1 of β2-Glycoprotein1 | purified β2-Glycoprotein1 |
| Conjugate | Isoluminol conjugated anti-human IgG | HRP conjugated anti-human IgG |
| Calibration | Lot specific Master Curve + twoCalibrators (included in kit) | Five lot specific calibrators(Included in the kit) |
| Units | < 20 CU Negative result≥ 20 CU Positive result | < 20 SGU Negative result≥ 20 SGU Positive result |
| AnalyticalMeasuring Range | 3.6 – 1380.4 CUReportable range = up to 13804.0 CU | 9.4 – 150.0 SGU |
| Sample type | Serum or citrated plasma | Serum |
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| Item | QUANTA Flash β2GP1-Domain1Calibrators | Predicate Device |
|---|---|---|
| Intended use | No separate intended use; calibratorsare part of the kit. | No separate intended use; calibratorsare part of the kit. |
| Analyte | Anti-β2GP1-Domain 1 antibodies | Anti-β2GP1 antibodies |
| Method | QUANTA Flash ® β2GP1-Domain1 | QUANTA Lite® β2 GPI IgG ELISA |
| Unit | Chemiluminescent Units (CU)QUANTA FlashArbitrary Units (U/ml)HemosIL AcuStar | SGU |
| Matrix | Human serum, stabilizers, andpreservative | Human serum, stabilizers, andpreservative |
| Physico-chemicalcharacteristics | Liquid, prediluted, ready to use | Liquid, prediluted, ready to use |
| Storage | 2-8 °C | 2-8 °C |
| Shelf life | One year | One year |
QUANTA Flash ß2GP1-Domain1 Calibrators
QUANTA Flash ß2GP1-Domain1 Controls
| Item | QUANTA Flash β2GP1-Domain1Controls | Predicate Device |
|---|---|---|
| Intended use | The QUANTA Flash β2GP1-Domain1Controls are intended for qualitycontrol purposes of the QUANTA Flashβ2GP1-Domain1 chemiluminescentimmunoassay (CIA) kit. | No separate intended use; controls arepart of the kit. |
| Analyte | Anti-β2GP1-Domain1 antibodies | Anti- β2GP1- antibodies |
| Method | QUANTA Flash β2GP1-Domain1chemiluminescent immunoassay | QUANTA Lite® β2 GPI IgG ELISA |
| Unit | QUANTA Flash (CU)HemosIL AcuStar (U/ml) | SGU |
| Matrix | Human serum, stabilizers, andpreservative | Human serum, stabilizers, andpreservative |
| Physico-chemicalcharacteristics | Liquid, ready to use | Liquid, prediluted, ready to use |
| Levels | 2 (low and high) | 2 (negative and positive) |
| Storage | 2-8°C | 2-8°C |
| Shelf life | One year | One year |
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Analytical performance characteristics
Value assignment and traceability of Calibrators and Controls
The QUANTA Flash ß2GP1-Domain1 Calibrators and Controls are manufactured by diluting human serum that contains high titer of anti-R2GP1-Domain1 antibody stabilizer buffer, containing preservative. The human serum is obtained from commercial sources and it is tested for markers of infectious substances.
The target CU is achieved through trial dilutions on small scale. Once a dilution is selected, the Calibrators and Control are bulked, tested, and adjusted. Upon completion of the manufacturing process, the Calibrators and Controls are tested on at least two instruments, on at least two lots of reagent cartridge, in replicates of 10 to determine final value assignment.
There are no international reference standards for anti- ß2GP1 IgG. Calibrators and controls are assigned values based on a 20 CU cut-off between positive (high) and negative (low) during assay development. Calibrators are specified in the labeling and supplied with the assay. Control materials are sold separately. The table below summarizes the control and calibrators target values.
| Points on master curve | Assigned Value (CU) |
|---|---|
| Point 1 | 3.6 |
| Point 2 | 18.5 |
| Point 3 | 43.7 |
| Point 4 | 90.0 |
| Point 5 | 316.0 |
| Point 6 | 1380.4 |
List of ß2GP1-Domain1 Standards, Calibrators and Controls:
| Material | ManufacturingTarget Value (CU) | Manufacturing TargetRange (CU) |
|---|---|---|
| Calibrator 1 | 19.0 | 17 – 21 |
| Calibrator 2 | 316.0 | 276 – 356 |
| Control Low | 10.0 | 8 – 12 |
| Control High | 50.0 | 40 – 60 |
Sample Matrix Comparison
Seventy-nine matched serum and citrated plasma samples (with different reactivity levels, 13.9% of samples around the cut-off) were tested side-by-side in singleton on QUANTA Flash® ß2GP1-Domain1 and analyzed according to CLSI Guideline EP09/A3.
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The user can select the type of sample matrix at the time of loading the samples onto the BIO-FLASH instrument. The choices on matrices are serum or plasma, and each of them is associated to a specific protocol that uses different sample dilution factors (1/10 for serum and 1/8.4 for plasma) to compensate for the addition of the anticoagulant (sodium citrated plasma sample collection. The BIO-FLASH assay protocol for serum and plasma is the same after the initial dilution step. The plasma dilution factor was chosen to be consistent with the rest of the APS assay family (QUANTA Flash aCL IgG and ß2GP1 IgG), where the dilution factor ratio between the serum protocol and the plasma protocol is 1.19 (in the Domain1, 10 / 8.4 = 1.19).
Percent difference and unit difference were calculated for each paired samples are considered equivalent if the percent difference between them is ≤15% or if the difference in units (CU) is ±3 CU (which is ≤15% of the cut-off). In addition, a linear regression comparison was performed and its acceptance criteria and results are summarized in the table below.
| Acceptance Criteria | Serum vs. Plasma | |
|---|---|---|
| Weighted R | $\geq$ 0.975 | 0.997 |
| Intercept of the regression line (Constant bias) | $\pm$ 3CU | 0.1 |
| Slope of the regression line (Proportional bias) | 0.9-1.1 | 1.01 |
| Weighted S y/x (Standard error of estimate) | $\leq$ 0.5 | 0.064 |
| Predicted bias (difference ) at cut-off | $\pm$ 3CU | 0.3 |
| 95% Cl of the bias | <20% $\pm$ 4CU | 0.00to 1.03 |
All paired samples fulfill the acceptance criteria, where 53/60 positive samples showed ≤10% difference, being 12.8% the maximum difference obtained. Therefore, the assay is claimed to be used with serum and citrated plasma.
Precision
The precision of the QUANTA Flash ß2GP1-Domain1 assay was evaluated on 8 serum samples containing various concentrations of anti-Domain1 antibodies in accordance with CLSI EPO5-A2, Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline. Samples were run in duplicates, twice a day, for 20 days.
Data were analyzed with the Analyse-it for Excel method evaluation software, and within run, between run, between day and total precision were calculated.
Acceptance criteria: Total %CV: < 15%
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| QUANTA Flash β2GP1-Domain1 Precision Study Decision Summary | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Within Run | Between-Run | Between-Day | Total | |||||||
| Sample | N | Mean (CU) | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Precision 1 | 80 | 10.4 | 0.9 | 8.3% | 0.0 | 0.0% | 0.5 | 4.3% | 1.0 | 9.4% |
| Precision 2 | 80 | 18.4 | 0.9 | 4.6% | 0.5 | 2.9% | 0.5 | 2.6% | 1.1 | 6.0% |
| Precision 3 | 80 | 22.8 | 1.4 | 6.1% | 0.9 | 3.8% | 0.0 | 0.0% | 1.6 | 7.2% |
| Precision 4 | 80 | 54.3 | 2.5 | 4.6% | 1.4 | 2.5% | 1.2 | 2.2% | 3.1 | 5.6% |
| Precision 5 | 80 | 99.6 | 6.4 | 6.5% | 0.0 | 0.0% | 0.0 | 0.0% | 6.4 | 6.5% |
| Precision 6 | 80 | 318.3 | 18.6 | 5.9% | 0.0 | 0.0% | 4.9 | 1.6% | 19.3 | 6.1% |
| Precision 7 | 80 | 538.1 | 30.3 | 5.6% | 7.9 | 1.5% | 28.7 | 5.3% | 42.5 | 7.9% |
| Precision 8 | 80 | 947.8 | 82.8 | 8.7% | 57.3 | 6.0% | 0.0 | 0.0% | 100.7 | 10.6% |
Results are summarized in the Table below.
Summary of Precision study QUANTA Flash
| No. of samples | Within Run | Between-Run | Between-Day | Total | |
|---|---|---|---|---|---|
| β2GP1-Domain1 | 8 | 4.6-8.7% | 0.0-6.0% | 0.0-5.3% | 5.6-10.6% |
Plasma Precision Verification
The precision of the QUANTA Flash B2GP1-Domain1 assay using plasma samples was verified using 3 plasma samples containing various concentrations of antibodies in accordance with CLSI EP15-A3, User Verification of Precision and Estimation of Bias; Approved Guideline - Third Edition. Samples were run in replicates of 5, once a day, for 5 days.
Data were analyzed with the Analyse-it for Excel method evaluation software, and within run, between day and total precision were calculated.
Acceptance criteria: Total %CV: < 15%
Results are summarized in the Table below.
| QUANTA Flash β2GP1-Domain1 Plasma Precision Verification Summary | ||||||||
|---|---|---|---|---|---|---|---|---|
| Within Run | Between-Day | Total | ||||||
| Sample | N | Mean | SD | %CV | SD | %CV | SD | %CV |
| Plasma Precision 1 | 25 | 444.6 | 15.0 | 3.4% | 7.9 | 1.8% | 16.9 | 3.8% |
| Plasma Precision 2 | 25 | 92.0 | 2.3 | 2.5% | 0.3 | 0.3% | 2.3 | 2.5% |
| Plasma Precision 3 | 25 | 18.7 | 0.9 | 4.6% | 1.2 | 6.2% | 1.5 | 7.8% |
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510(k) Summary QUANTA Flash® ß2GP1-Domain1
Page 11 of 19
Limit of Blank (LoB) and Limit of Detection (LoD)
The LoD of the QUANTA Flash 12GP1-Domain1 assay is 1296 RLU, which is below the analytical measuring range of the assay. It was determined consistent with CLSI EP17-A2 guideline with proportions of false positives (alpha) less than 5% and false negatives (beta) less than 5%; based on 140 determinations, with 60 measurements on blank samples and 80 measurements of low level samples. The LoB is 425.7 RLU.
These values are below the value of the lowest QUANTA Flash ß2GP1-Domain1 Master Curve standard, i.e. below the Analytical Measuring Range.
Analytical Measuring Range (AMR)
QUANTA Flash ß2GP1-Domain1: 3.6 CU - 1380.4 CU
The AMR is defined by the values of the lowest and highest Master Curve Standards.
Auto-rerun function and reportable results
The BIO-FLASH software has an Auto-rerun option available. If this option is selected, the instrument will automatically rerun any sample that has a result >1380.4 CU by further diluting it by a factor specified in the assay definition file (10 fold), thereby bringing the measured value within the AMR. The final result will be calculated by the software by taking into account the additional dilution factor. As the highest value that can be measured is 1380.4 CU, the highest value that can be reported is 13804 CU.
To validate the Auto-rerun function, three high positive specimens with results above the analytical measuring range were selected. The samples were run with the Auto-rerun function enabled on the BIO-FLASH. Then the specimens were manually diluted the same way as it happens in the Auto-rerun function (10 fold), and tested on the BIO-FLASH. The results were within the analytical measuring range after autorerun or manual dilution for all specimens. The % recovery values for results obtained with the auto-rerun results compared to the results obtained by manual dilution were between 91.9% and 99% (average 94.9%).
High concentration hook effect
To assess hook effect, measurement signal (RLU) was examined by performing serial dilutions of two high positive samples (with results above the AMR when tested as neat samples). RLU values showed increase with increasing antibody concentrations above the AMR, thereby confirming that high positive specimens above the analytical measuring range do not show hook effect up to 16,060.8 CU (theoretical value calculated using the highest value in the AMR and its dilution factor) in the ß2GP1-Domain1 assay.
Linearity
The linearity of the AMR was evaluated by a study according to CLSI EPG-A, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approved Guideline. Five serum samples and three plasma samples with various ß2GP1-Domain1 antibody concentrations were diluted with negative serum or plasma in 10% increments (from 0% to 90% negative serum) to obtain values that cover the AMR. The dilutions were assayed in duplicates. Percent recovery of obtained mean results was
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510(k) Summary QUANTA Flash® ß2GP1-Domain1
calculated compared to the expected mean results (based on the dilution factor). Moreover, obtained values of individual replicates were plotted against expected values, and linear regression analysis was performed.
-
Recovery is between 80-120%, or ± 4 CU, whichever is greater. Acceptance criteria:
-
For linear regression analysis, slope is between 0.9-1.1, and R2 is ≥ 0.95.
All five serum specimens showed dilution linearity individually and combined.
| Sample | Test Range (CU) | Slope (95% CI) | R2 | % Recovery Range |
|---|---|---|---|---|
| 1 | 1640.7 to 164.1 | 1.04 (0.98 - 1.09) | 0.99 | 95.1%-106.2% |
| 2 | 454.7 to 45.5 | 0.99 (0.96 - 1.03) | 0.99 | 95.5%-104.1% |
| 3 | 124.9 to 12.5 | 1.03 (1.00 - 1.05) | 1.00 | 94.3%-103.3% |
| 4 | 81.1 to 8.1 | 1.01 (0.95 - 1.08) | 0.98 | 83.9%-106.7% |
| 5 | 10.3 to 1.0 | 0.96 (0.91 - 1.01) | 0.99 | 90.9%-113.9% |
| Combined | 1640.7 to 1.0 | 1.01 (0.99 to 1.02) | 1.00 | 83.9%-113.9% |
All three plasma specimens showed dilution linearity individually and combined.
| Plasma | Test Range (CU) | Slope (95% CI) | R² | % Recovery Range |
|---|---|---|---|---|
| 1 | 1,357.6 to 135.8 | 1.02 (0.96 - 1.07) | 0.99 | 86.6%- 102.6% |
| 2 | 182.2 to 18.2 | 0.97 (0.93 - 1.01) | 0.99 | 90.1%- 100.0% |
| 3 | 17.7 to 3.5 | 0.96 (0.93 - 0.99) | 1.00 | 98.2%- 111.9% |
| Combined | 1,357.6 to 3.5 | 0.96 (0.94 - 0.98) | 0.99 | 86.6%- 111.9% |
These data demonstrate the linearity of the analytical measuring range (3.6 CU - 1380.4 CU) of the QUANTA Flash ß2GP1-Domain1 assay using serum or plasma samples.
Interference
The interference study was performed according to CLSI EPO7-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition. Five specimens were tested, three serum samples (negative: 10.7 CU; low positive: 55.9 CU; high positive: 456.6 CU) and two plasma samples (negative: 6.6 CU; high positive: 132.4 CU). Interfering substances (bilirubin, hemoglobin, triglycerides and cholesterol) were spiked into every specimen at three different concentrations in 10% of total specimen volume, and the resulting samples were assessed in triplicates with the ß2GP1-Domain1 assay. For Rheumatoid Factor lgM, the samples are diluted to varying extent depending on the desired concentrations of RF as an interferent, and the resulting samples were assessed in triplicates with the ß2GP1-Domain1 assay. Recovery of the unit values was calculated compared to control samples. Acceptance criteria for the interference studies were 85%-115% recovery, or ±4 CU difference, whichever is greater.
No interference was detected with bilirubin up to 100 mg/dL (recovery: 92.9% to 109.4%), hemoglobin up to 200 mg/dL (recovery: 95.6% to 106.3% or 1.7 CU), triglycerides up to 1000 mg/dL (recovery: 92.5% to 107.7%), cholesterol up to 224.3 mg/dL (recovery: 92.5% to 107.7%), and RF IgM up to 500 IU/mL (recovery: 86.2% to 109.9% or 0.9 CU).
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Cross-reactivity
A potential cross-reactivity of the QUANTA Flash ß2GP1 Domain1 CIA with other autoantibodies was evaluated with 613 control samples that were included in the clinical validation study. These samples were from patients with autoimmune diseases that are characterized with disease specific autoantibodies, or from patients with infection. The composition of the cohort and the anti-ß2GP1 Domain1 positivity rate is shown in the Table below:
| Diagnosis | Number of samples | # pos | % pos |
|---|---|---|---|
| Crohn's Disease | 104 | 1 | 1.0% |
| Ulcerative Colitis | 94 | 0 | 0.0% |
| Rheumatoid Arthritis | 168 | 0 | 0.0% |
| Osteoarthritis | 49 | 0 | 0.0% |
| Scleroderma | 127 | 2 | 1.6% |
| Hepatitis B virus | 21 | 0 | 0.0% |
| Hepatitis C virus | 10 | 0 | 0.0% |
| Syphilis | 40 | 0 | 0.0% |
| Total controls | 613 | 3 | 0.5% |
Based on the results, the QUANTA Flash ß2GP1 Domain1 assay does not show cross-reactivity with autoantibodies that are present in various autoimmune diseases or with antibodies against infectious agents.
Lot to lot comparison
Twenty-six samples with various reactivity levels were tested with three different reagent lots: RP0003 (RP3), RP011 (RP11) and 131001P1 (PP1). The samples covered the total analytical measuring range of the assay. Results were processed by linear regression analysis and bias calculation according to CLSI EPO9-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples; Approved Guideline. Pair-wise comparisons were performed between lot RP3 vs RP11 and lot RP11 vs PP1.
Acceptance criteria and results are in the Table below. All results were within the acceptance limits.
| Acceptance criteria | RP3 vs RP11 | RP11 vs PP1 | PP1 vs RP3 | |
|---|---|---|---|---|
| Weighted R for linear regression | $\geq 0.975$ | 0.996 | 0.998 | 0.986 |
| Intercept of the regression line (constant bias) | $\pm 3$ CU | 1.9 | 0.06 | -1.3 |
| Slope of the regression line (proportional bias) | 0.9 - 1.1 | 1.03 | 1.04 | 1.00 |
| Weighted S y/x (Standard error estimate) | $\leq 0.5$ | 0.07 | 0.05 | 0.13 |
| Predicted bias (difference) at cut-off | $\pm 3$ CU | 2.5 | 0.8 | -1.2 |
| 95% CI of the bias | $\leq 20%$ or $\pm 4$ CU | 1.1 to 3.8 | -0.3 to 1.8 | -3.9 to 1.6 |
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Stability
Shelf life
To establish the initial claim for shelf life, accelerated stability studies were performed for 4 weeks at 37 °C, where one week is equal to six months at 5 ± 3°C.
Accelerated stability testing was performed on each of the following sealed components of the QUANTA Flash® ß2GP1-Domain1 to establish initial stability claim:
| • QUANTA Flash® β2GP1-Domain1 Reagent Kit | (1 Lot) |
|---|---|
| • β2GP1-Domain1 beads | (3 Lots) |
| • Resuspension Buffer | (3 Lots) |
| • Calibrators 1 and 2 | (3 Lots) |
| • Low and High controls | (3 Lots) |
Each week a new sealed component was placed in the incubator, and all components were tested at the end of the experiment together with the one that was stored at 5 ± 3℃. The recovery of the measured values was calculated for each time point (compared to those obtained with 5 ± 3°C stored reagent). All calculations were performed by comparing results of sealed components stored at 5 ± 3℃ (control) to those stored at 37 ± 3℃ (test) for 1, 2, 3, and 4 weeks, where one week is equal to six months at 5 ± 3℃. Linear regression analysis was performed between recovery values and the number of days.
Acceptance criteria for one year preliminary expiration dating:
- Beads, Resuspension Buffer, and Reagent Kit:
With regression analysis, the lower and upper 95% Cl interval of the regression line is between 85% and 115% recovery at day 14, and no individual data point has ≤75% or ≥125% recovery at day 14.
- Controls and Calibrators:
With regression analysis, the lower and upper 95% Cl interval of the regression line is between 90% and 110% recovery at day 14, and no individual data point has ≤80% or ≥120% recovery at day 14.
All components tested fulfilled the acceptance criteria above, so one year expiration dating was assigned to each component
In-use (onboard) stability
Calibrators
Onboard stability claim: 4 calibrations, or 8 hours onboard
During assessment of on-board stability, Calibrators were placed uncapped, onboard the instrument, and calibration was performed altogether five times over 8.5 hours. Controls and a panel of characterized patient specimens were run on each calibration curve.
Calibrators are considered stable if all five calibrations performed in the 8.5 hour period are successful, and average Calibrator RLU recovery values are between 90% and 110% compared to the first use.
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A total of 5 successful calibrations were performed over a 8.5 hour period. Calibrator RLU values remained within the 90-110% range. Moreover, all Controls and patient panel samples ran within their expected range. This supports the claim that calibrators can be used for up to 4 calibrations over an 8 hour period.
Controls
Onboard stability claim: up to 15 uses, at 10 minutes onboard per use
During assessing on-board stability, 2 vials of each Control were assayed twice a day for a total of 20 runs. The first run was used to establish baseline value, by running each vial in duplicate, and then additional 19 runs were performed, by running each vial in singleton. During runs, the Controls were left uncapped, onboard the instrument for 15 minutes per run. When not in use, the controls were capped, and stored at 5 ± 3°C. Percent recovery of each value was calculated compared to the baseline value. Controls are considered stable when all values run within their established range, and the linear regression line obtained by plotting %recovery values against the number of runs stays between 85% and 15. All controls ran within their respective acceptable ranges for all runs. Moreover, the regression line remained between 85% and 115% at run 15 for both Controls. These results support the claim that controls can be used for up to 15 times, at 10 minutes per use.
Reagent Cartridge
To establish the in-use (onboard) stability of the QUANTA Flash ß2GP1-Domain1 reagent cartridge, three lots of reagent cartridge were tested with up to 4 serum specimens (with different reactivity levels) along with the Low and High Controls. The specimens were tested periodically up to 64 days. Percent recoveries were calculated compared to the day zero average values, and linear regression analysis was performed by plotting %recovery against the number of days. The claim was established using the following criteria (using the one that is fulfilled first):
-
The stability claim is established at the actual measurement day preceding the day when the 95% confidence interval of the regression line reaches 85% or 115% recovery, or
-
At the actual measurement day preceding the day when 2 data points or ≥2% of the recovery data (whichever is greater) is ≤ 75% or ≥ 125% recovery.
As none of these endpoints were reached during the duration of the in-use (onboard) stability of Domain1 reagent cartridge was set at 60 days.
Real time stability
Real time stability testing has been scheduled to be performed approximately every three months on the reagent cartridge, Calibrators and Controls, to verify the one year expiration that was assigned based on accelerated stability studies. At the of the submission, results were available up to 12 months for reagent cartridge, up to 15 months on Calibrators and Controls.
For reagent cartridge, QC panel samples were tested in singleton at each time point. The QC panel is a group of characterized patient samples with target values, used by the QC Department for reagent release and QC.
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- Acceptance criteria: results should fall within their respective QC ranges.
Calibrators were used to calibrate a cartridge at each time point. After calibration, the QC panel samples were tested in singleton at each time point.
- Acceptance criteria: results should fall within their respective QC ranges.
Controls were tested in singleton on a calibrated cartridge at each time point. Individual values were compared to the values that were assigned to the Controls at release.
- Acceptance criteria: results should fall within their acceptable ranges as were established at the release of the Controls.
All results to date were within the acceptance limit, therefore one year expiration dating has been verified through real-time studies.
Sample stability
Sample stability claim: -Samples can be stored at room temperature for up to 48 hours.
-Samples can be stored at 2-8°C for up to 14 days.
-Samples can be frozen and thawed up to 3 times.
During the sample stability study, eight (four serum and four plasma samples) samples with varying anti-Domain1 levels have been tested along with Low and High Controls at different time points after being stored at each temperature condition. For the room temperature claim, samples were tested at 0, 8, 24 and 48 hours. For the 2-8°C claim, samples were tested at 0, 1, 2, 3, 7 and 14 days. For the freeze and thaw claim, samples were tested after 0, 1, 2 and 3 cycles. Samples were tested in duplicates at each time point. Percent recoveries were calculated at each time point compared to the time zero average values. Samples are considered stable if the recovery is between 85%-115% for positive samples. Samples are considered stable if the recovery is between 80-120%, or 20% of the cutoff in units (4 CU) for negative samples.
This study has been performed using serum and citrated plasma samples and all results fulfilled the acceptance criteria above, so the following claims have been established for both serum and citrated plasma:
-Samples can be stored at room temperature for up to 48 hours.
-Samples can be stored at 2-8°C for up to 14 days.
-Samples can be frozen and thawed up to 3 times.
Cut-off, reference range
| QUANTA Flash ß2GP1-Domain1: | Negative | <20 CU |
|---|---|---|
| Positive | ≥20 CU |
The reference population for establishing cut-off for the ß2GP1-Domain1 assay consisted of 30 subjects:
| Sample Type | Number of Samples |
|---|---|
| Apparently healthy blood donors | 5 |
| Viral hepatitis | 4 |
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| Sample Type | Number of Samples |
|---|---|
| Systemic Lupus erythematosus (withouthistory of thrombotic events) | 10 |
| Syphilis | 10 |
| HIV | 1 |
All specimens were the same matrix (Serum). All specimens were unaltered. The cut-off was established in accordance to CLSI EP28-A3c: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline - Third Edition. The Analyse-it for Excel software was used to make the calculations. The 99th percentile of the obtained values was calculated as 7866 RLU.
The cutoff was set to 7880 RLU, which was set to equal 20 CU. One Systemic Lupus erythematosus (without history of thrombotic events) patient tested positive at this cutoff level.
Clinical performance characteristics
Clinical sensitivity, specificity
A cohort of 1090 characterized samples, none of which were used for establishing the reference range, was used to validate the clinical performance of the QUANTA Flash ß2GP1-Domain1 CIA. All samples were run on the QUANTA Flash ß2GP1-Domain1. The distribution of the cohort and the ß2GP1-Domain1 positivity rate is in the Table below:
| Cohort | N=1090 | No (%) positive |
|---|---|---|
| APS combined | 270 | 138 (51.1%) |
| pAPS | 180 | 102 (56.6%) |
| sAPS | 90 | 36 (40.0%) |
| Infectious diseases | 71 | 0 (0.0%) |
| Hepatitis B virus (HBV) | 21 | 0 (0.0%) |
| Hepatitis C virus (HCV) | 10 | 0 (0.0%) |
| Syphilis | 40 | 0 (0.0%) |
| Other diseases | 566 | 3 (0.5%) |
| Crohn's Disease (CD) | 104 | 1 (1.0%) |
| Ulcerative Colitis (UC) | 94 | 0 (0.0%) |
| Rheumatoid Arthritis (RA) | 168 | 0 (0.0%) |
| Osteoarthritis (OA) | 49 | 0 (0.0%) |
| Scleroderma | 127 | 2 (1.6%) |
| Others | 24 | 0 (0.0%) |
| Conditions "without APS" | 183 | 0 (0.0%) |
| Pre-eclampsia/eclampsia | 34 | 0 (0.0%) |
| Fetal loss no APS | 45 | 0 (0.0%) |
| SLE no APS | 37 | 0 (0.0%) |
| Thrombosis no APS | 41 | 0 (0.0%) |
| Atopic Dermatitis | 26 | 0 (0.0%) |
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Clinical sensitivity and specificity for APS (n=270) using the control population (n=820) is calculated in the table below.
| Cohort 1 (n=1090) | Diagnosis | Percent Agreement(95% confidence) | |||
|---|---|---|---|---|---|
| APS | Controls | Total | |||
| QUANTA Flash®Domain 1 CIA | Positive | 138 | 3 | 141 | Sensitivity = 51.1% (45.0-57.2%) |
| Negative | 132 | 817 | 949 | Specificity = 99.6% (98.9-99.9%) | |
| Total | 270 | 820 | 1090 |
To assess diagnostic efficiency, ROC analysis was performed on the validation sample pool for APS. The results are below:
| Test | Area | 95% CI | SE | Z | p |
|---|---|---|---|---|---|
| QUANTA Flash B2GP1-Domain1(CU) | 0.84 | 0.81 to 0.86 | 0.015 | 22.24 | <0.0001 |
Expected values
The expected value in the normal population is "negative". Anti-ß2GP1-Domain1 antibody levels were analyzed in a cohort of 400 apparently healthy blood donors (191 females, ages 17 to 60 years, average age 32.3 years, and 209 males ages 17 to 60 years, average age 34.7 years) using the QUANTA Flash ß2GP1-Domain1. This patient population was different from the one that was used to establish or validate the cutoff, and was only used to assess expected values. With a cut-off of 20 CU, one sample (0.25%) was positive (34.2 CU) on the QUANTA Flash (3GP1-Domain1. The mean concentration was 3.8 CU, and the values ranged from <3.6 to 34.2 CU.
Comparison with predicate device
Samples for method comparison analysis included all samples that fell within the reportable range from the clinical validation study, along with 8 additional samples, for a total of 238 samples. These additional samples were contrived by diluting ß2GP1-Domain1 positive samples with negative serum in order to increase the number of samples around the cut-off. All samples were tested on the QUANTA Flash " ß2GP1-Domain1 CIA and the QUANTA Lite ß›GPI IgG ELISA, which is used as a predicate device. Out of the 238 total samples, 22 samples (9.24%) were around the cut-off (15.1 to 25.2 CU).
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510(k) Summary QUANTA Flash® ß2GP1-Domain1
| All (n=238) | QUANTA Lite® β2GPI IgG ELISA | Percent Agreement (95% confidence) | ||||
|---|---|---|---|---|---|---|
| Positive | Negative | Total | ||||
| QUANTA Flash®β2GP1-Domain1 CIA | Positive | 101 | 28 | 129 | Pos. Agree = 91.0% (84.1-95.6%) | |
| Negative | 10 | 99 | 109 | Neg. Agree = 78.0% (69.7-84.8%) | ||
| Total | 111 | 127 | 238 | Total Agree = 84.0% (78.7-88.4%) |
Method Comparison, samples within reportable range:
There are 28 samples that reported a positive result in the new device but negative in the predicate device, 26/28 (92.9%) are from samples with an APS diagnosis and 2/28 (7.1%) are controls. There are 10 samples reported a negative result in the new device but positive in the predicate device, 5/10 (50.0%) are from control samples and 5/10 (50.0%) from APS samples.
Method Comparison, samples around the cut-off:
| All (n=22) | QUANTA Lite® β₂GPI IgG ELISA | Percent Agreement (95% confidence) | |||
|---|---|---|---|---|---|
| Positive | Negative | Total | |||
| QUANTA Flash®β2GP1-Domain1 CIA | Positive | 4 | 3 | 7 | Pos. Agree = 80.0% (28.4-99.5%) |
| Negative | 1 | 14 | 15 | Neg. Agree = 82.4% (56.6-96.2%) | |
| Total | 5 | 17 | 22 | Total Agree = 81.8% (59.7-94.8%) |
§ 866.5660 Multiple autoantibodies immunological test system.
(a)
Identification. A multiple autoantibodies immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoantibodies (antibodies produced against the body's own tissues) in serum and other body fluids. Measurement of multiple autoantibodies aids in the diagnosis of autoimmune disorders (disease produced when the body's own tissues are injured by autoantibodies).(b)
Classification. Class II (performance standards).