(258 days)
Not Found
No
The description details a standard automated immunoassay system and its components, with no mention of AI or ML algorithms for data analysis or interpretation. The results are calculated based on a predefined master curve and instrument-specific working curve, which is a traditional method for quantitative assays.
No
This device is an immunoassay intended for in vitro diagnostic use to aid in the diagnosis of certain autoimmune diseases by detecting specific autoantibodies, not for therapeutic purposes.
Yes
The "Intended Use / Indications for Use" section states that the device is "an aid in the diagnosis of Systemic Lupus Erythematosus, Systemic Sclerosis, Idiopathic Inflammatory Myopathies."
No
The device description explicitly states that the assay runs on the BIO-FLASH instrument, which is described as a "fully automated closed system" including "liquid handling hardware, luminometer and computer with software-user interface." The assay also utilizes physical reagent cartridges, beads, and buffers. This indicates the device is a combination of hardware, software, and reagents, not a software-only medical device.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it is for the "semi-quantitative determination of IgG anti-Ro52 autoantibodies in human serum." This involves testing a sample taken from the human body (serum) to provide information for the diagnosis of specific conditions (Systemic Lupus Erythematosus, Systemic Sclerosis, Idiopathic Inflammatory Myopathies). This aligns directly with the definition of an in vitro diagnostic device.
- Device Description: The description details a laboratory assay that analyzes a biological sample (serum) using chemical and immunological reactions to measure a specific analyte (IgG anti-Ro52 autoantibodies). This process is performed in vitro (outside the body).
- Performance Studies: The document includes performance studies evaluating the device's clinical sensitivity and specificity in relation to specific diseases, which is a requirement for IVDs.
- Predicate Device: The mention of a predicate device (K063565; QUANTA Lite® SS-A 52 ELISA) further confirms its classification as an IVD, as predicate devices are used for comparison in the regulatory submission process for new IVDs.
N/A
Intended Use / Indications for Use
QUANTA Flash Ro52 is a chemiluminescent immunoassay for the semi-quantitative determination of IgG anti-Ro52 autoantibodies in human serum. The presence of anti-Ro52 autoantibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of Systemic Lupus Erythematosus, Sjögren's Syndrome, Systemic Sclerosis, Idiopathic Inflammatory Myopathies.
QUANTA Flash Ro52 Calibrators are intended for use with the QUANTA Flash Ro52 Reagents for the determination of IgG anti-Ro52 autoantibodies in human serum. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash Ro52 Controls are intended for use with the QUANTA Flash Ro52 Reagents for quality control in the determination of IgG anti-Ro52 autoantibodies in human serum.
Product codes (comma separated list FDA assigned to the subject device)
OBE, JIT, JJX
Device Description
The QUANTA Flash Ro52 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 Ro52 assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH instrument.
Purified recombinant Ro52 antigen is coated onto paramagnetic beads. The bead suspension is lyophilized and stored in the bead tube. Prior to use in the BIO-FLASH system, the sealed reagent tubes are pierced with the reagent cartridge lid and the beads are rehydrated and resuspended using resuspension buffer by pipetting up and down with a transfer pipette. The reagent cartridge is then loaded onto the BIO-FLASH instrument. Samples are also loaded onto the instrument in sample racks. Serum samples are prediluted 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. Isoluminol 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(II)coproporphyrin in sodium hydroxide solution) and Trigger 2 (ureahydrogen 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-Ro52 antibodies bound to the corresponding beads.
For quantitation, the QUANTA Flash Ro52 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 Ro52 Calibrators. Based on the results obtained with the two Calibrators included in the Calibrator Set (sold separately), 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 Ro52 kit contains the following materials:
One (1) QUANTA Flash Ro52 Reagent Cartridge
One (1) vial of Resuspension buffer
One (1) Transfer pipette
The QUANTA Flash Ro52 reagent cartridge contains the following reagents for 50 determinations:
- a. Ro52 antigen coated paramagnetic beads, lyophilized.
- Assay buffer colored pink, containing Tris-buffered saline, Tween 20, protein b. stabilizers and preservatives.
- C. Tracer IgG - Isoluminol labeled anti-human IgG antibodies in buffer, containing protein stabilizers and preservative.
The QUANTA Flash Ro52 Calibrators kit contains two vials of Calibrator 1 and two vials of Calibrator 2:
QUANTA Flash Ro52 Calibrators:
- QUANTA Flash Ro52 Calibrator 1: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
- QUANTA Flash Ro52 Calibrator 2: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
The QUANTA Flash Ro52 Controls kit contains two vials of Negative Control and two vials of Positive Control:
QUANTA Flash Ro52 Controls:
- QUANTA Flash Ro52 Negative Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
- QUANTA Flash Ro52 Positive Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Not Found
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
A cohort of characterized samples, none of which were used for establishing the reference range, was used to validate the clinical performance of the QUANTA Flash Ro52. A total of 600 characterized samples were included in the Validation Set for the QUANTA Flash Ro52. All samples were run on the QUANTA Flash Ro52. The distribution of the Ro52 positivity rate is in the Table below:
Patient group, N, Number positive, % positive
Graves' Disease, 10, 0, 0.0%
Hashimoto Thyroiditis, 10, 0, 0.0%
Celiac Disease, 11, 0, 0.0%
Crohn's Disease, 20, 0, 0.0%
Ulcerative Colitis, 20, 0, 0.0%
HCV, 6, 1, 16.7%
HBV, 6, 0, 0.0%
CMV, 11, 1, 9.1%
EBV (with or without other infection), 12, 2, 16.7%
HIV, 5, 0, 0.0%
Syphilis, 5, 0, 0.0%
Primary Antiphospholipid Syndrome, 15, 0, 0.0%
Secondary Antiphospholipid Syndrome*, 14, 6, 42.9%
Vasculitis, 17, 0, 0.0%
Rheumatoid arthritis, 50, 2, 4.0%
Osteoarthritis, 20, 1, 5.0%
Behçet's disease, 1, 0, 0.0%
Total controls, 233, 13, 5.6%
Sjögren's Syndrome (SS), 91, 40, 44.0%
Systemic Lupus Erythematosus (SLE), 131, 47, 35.9%
Systemic Sclerosis (SSc), 80, 13, 16.3%
Idiopathic Inflammatory Myopathies (IIM), 65, 26, 40.0%
Total, 600, Not Found, Not Found
* Patients may have SLE
The results were analyzed to calculate sensitivity and specificity for SS (n=91), SLE (n=80), and IIM (n=65) separately, using 219 disease controls. The secondary APS group was excluded from all calculations.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision Study:
Study Type: Precision
Sample Size: 9 samples with various concentrations, run in duplicates, twice a day, for 21 days for a total of 84 replicates per sample.
Key Results: Total %CV was less than 10% for all samples, ranging from 5.5% to 8.5%.
Reproducibility Study:
Study Type: Reproducibility
Sample Size: 3 samples tested on two different reagent lots, using two different lots of Calibrators, by two operators. Samples were run in quadruplicates, two times a day, for 10 days, to generate 80 data points per sample.
Key Results: Total %CV was less than 10% for all samples, ranging from 4.3% to 6.7%.
Limit of Blank (LoB) and Limit of Detection (LoD) Study:
Study Type: Limit of Blank and Limit of Detection
Sample Size: LoB: 4 blank samples run in replicates of five on two reagent lots, once per day, for 3 days (60 data points per lot). LoD: 4 low level samples run in replicates of five on two reagent lots, once per day, for 3 days (60 data points per lot).
Key Results: LoB for lot 131006 was 308 RLU, and for lot 141007 was 319 RLU. The final LoB value is 319 RLU. LoD for lot 131006 was 400 RLU, and for lot 141007 was 402 RLU. The final LoD value is 402 RLU. These values are below the Analytical Measuring Range.
High concentration hook effect study:
Study Type: Hook effect
Sample Size: Five high positive samples.
Key Results: High positive specimens above the analytical measuring range do not show hook effect up to 23005 CU in the Ro52 assay (the highest concentration that was tested).
Linearity Study:
Study Type: Linearity
Sample Size: Four serum samples with various Ro52 antibody concentrations, diluted in 10% increments and assayed in duplicates.
Key Results: All four specimens showed dilution linearity individually. For combined data, the slope was 0.97 (0.96 to 0.97) and R-squared was 1.00.
Interference Study:
Study Type: Interference
Sample Size: Three specimens (negative, at cutoff, positive), spiked with interfering substances at three different concentrations, assessed in triplicates.
Key Results: No interference was detected with bilirubin up to 10 mg/dL, hemoglobin up to 200 mg/dL, triglycerides up to 1000 mg/dL, cholesterol up to 224.3 mg/dL, and RF IgM up to 500 IU/mL, as recoveries were within 85%-115% or ±4 CU difference.
Cross-reactivity Study:
Study Type: Cross-reactivity
Sample Size: 199 control samples from patients with various autoimmune diseases or positive infectious disease serology.
Key Results: Based on the results, the QUANTA Flash Ro52 assay does not show cross-reactivity with autoantibodies that are present in various autoimmune diseases, or antibodies against infectious agents.
Lot to lot comparison Study:
Study Type: Lot to lot comparison
Sample Size: Twenty unique samples and the Positive and Negative Controls (22 specimens) tested in triplicates with three different reagent lots.
Key Results: All results were within the acceptance limits for weighted r (>=0.975), intercept of regression line (±15% of cut-off), slope of regression line (0.9-1.1), weighted S y/x (
§ 866.5100 Antinuclear antibody immunological test system.
(a)
Identification. An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).(b)
Classification. Class II (performance standards).
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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
March 5. 2015
INOVA DIAGNOSTICS, INC. DR. GABRIELLA LAKOS DIRECTOR, RHEUMATOLOGY RESEARCH 9900 OLD GROVE ROAD SAN DIEGO, CA 92131-1638
Re: K141655 Trade/Device Name: Quanta Flash® Ro52 Ouanta Flash® Ro52 Calibrators Quanta Flash® Ro52 Controls Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear antibody immunological test system Regulatory Class: II Product Code: OBE, JIT, JJX Dated: January 15, 2015 Received: January 22, 2015
Dear Dr. Lakos:
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 of 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
1
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/Resourcesfor You/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,
Leonthena R. Carrington -A
Leonthena R. Carrington, MS, MBA, MT(ASCP) Director (Acting) 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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Indications for Use
510(k) Number (if known) K141655
Device Name
QUANTA Flash® Ro52, QUANTA Flash® Ro52 Controls, QUANTA Flash® Ro52 Calibrators,
Indications for Use (Describe)
QUANTA Flash Ro52 is a chemiluminescent immunoassay for the semi-quantitative determination of IgG anti-Ro52 autoantibodies in human serum. The presence of anti-Ro52 autoantibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of Systemic Lupus Erythematosus, Systemic Sclerosis, Idiopathic Inflammatory Myopathies.
QUANTA Flash Ro52 Controls are intended for use with the QUANTA Flash Ro52 Reagents for quality control in the determination of IgG anti-Ro52 autoantibodies in human serum.
QUANTA Flash Ro52 Calibrators are intended for use with the OUANTA Flash Ro52 Reagents for the determination of Ig G anti-Ro52 autoantibodies in human serum. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
| 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) | ☑ Prescription Use (Part 21 CFR 801 Subpart D) | ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) | ||
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary
QUANTA Flash® Ro52 QUANTA Flash® Ro52 Calibrators QUANTA Flash® Ro52 Controls
Table of Contents
| Administrative data | |
|---|---|
| Device description | |
| Intended use(s) | |
| Substantial equivalence | |
| Comparison to predicate device | |
| Analytical performance characteristics | |
| Value assignment and traceability of Calibrators and Controls | |
| Precision | |
| Reproducibility | |
| 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, Inc
9900 Old Grove Road,
San Diego, CA, 92131 |
|------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Purpose of submission: | New device(s) |
| Devices in the submission: | QUANTA Flash® Ro52
QUANTA Flash® Ro52 Calibrators
QUANTA Flash® Ro52 Controls |
| Scientific contact: | Gabriella Lakos, Director of Research and Development
Inova Diagnostics, Inc.
9900 Old Grove Road, San Diego, CA, 92131
Phone: 858-586-9900/1393
Fax: 858-863-0025
email: glakos@inovadx.com |
| Quality Systems contact: | Ronda Elliott, VP, Quality Systems and RA
Inova Diagnostics, Inc
9900 Old Grove Road, San Diego, CA, 92131
Phone: 858-586-9900/1381
Fax: 858-863-0025
email: relliot@inovadx.com |
| Preparation date: | 06/09/2014 |
| Device name (assay kit): | Proprietary name: QUANTA Flash® Ro52
Common name: Anti-Ro52 Chemiluminescent Immunoassay
Classification name: anti-SS-A 52 antibody, antigen and control |
| Regulation Description | Antinuclear antibody immunological test system |
| Regulation Medical Specialty | Immunology |
| Review Panel | Immunology |
| Product Code | OBE, Anti-SS-A 52 autoantibodies |
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510(k) Summary QUANTA Flash® Ro52
| Regulation Number | 866.5100 | |
|---|---|---|
| Device Class | 2 | |
| Device name (Calibrators): | Proprietary name: | QUANTA Flash® Ro52 Calibrators |
| Common name: | Ro52 Calibrators | |
| Classification name: | Calibrator, secondary | |
| Regulation Description | Calibrator | |
| Regulation Medical Specialty | Clinical Chemistry | |
| Product Code | JIT | |
| Regulation Number | 862.1150 | |
| Device Class | 2 | |
| Device name (Controls): | Proprietary name: | QUANTA Flash® Ro52 Controls |
| Common name: | Ro52 Controls | |
| Classification name: | single (specified) analyte controls (assayed and unassayed) | |
| 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: | ||
| Device description | QUANTA Lite® SS-A 52 ELISA, 510(k) number: K063565 |
The QUANTA Flash Ro52 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 Ro52 assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH instrument.
Purified recombinant Ro52 antigen is coated onto paramagnetic beads. The bead suspension is lyophilized and stored in the bead tube. Prior to use in the BIO-FLASH system, the sealed reagent tubes are pierced with the reagent cartridge lid and the beads are rehydrated and resuspended using
6
resuspension buffer by pipetting up and down with a transfer pipette. The reagent cartridge is then loaded onto the BIO-FLASH instrument. Samples are also loaded onto the instrument in sample racks. Serum samples are prediluted 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. Isoluminol 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(II)coproporphyrin in sodium hydroxide solution) and Trigger 2 (ureahydrogen 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-Ro52 antibodies bound to the corresponding beads.
For quantitation, the QUANTA Flash Ro52 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 Ro52 Calibrators. Based on the results obtained with the two Calibrators included in the Calibrator Set (sold separately), 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 Ro52 kit contains the following materials:
One (1) QUANTA Flash Ro52 Reagent Cartridge One (1) vial of Resuspension buffer
One (1) Transfer pipette
The QUANTA Flash Ro52 reagent cartridge contains the following reagents for 50 determinations:
- a. Ro52 antigen coated paramagnetic beads, lyophilized.
- Assay buffer colored pink, containing Tris-buffered saline, Tween 20, protein b. stabilizers and preservatives.
- C. Tracer IgG - Isoluminol labeled anti-human IgG antibodies in buffer, containing protein stabilizers and preservative.
The QUANTA Flash Ro52 Calibrators kit contains two vials of Calibrator 1 and two vials of Calibrator 2:
QUANTA Flash Ro52 Calibrators:
- । QUANTA Flash Ro52 Calibrator 1: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
- QUANTA Flash Ro52 Calibrator 2: Two (2) barcode labeled tubes containing 0.3 mL -
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prediluted, ready to use reagent. Calibrators contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
The QUANTA Flash Ro52 Controls kit contains two vials of Negative Control and two vials of Positive Control:
QUANTA Flash Ro52 Controls:
- । QUANTA Flash Ro52 Negative Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
- , QUANTA Flash Ro52 Positive Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to Ro52 in buffer, protein stabilizer and preservative.
Intended use(s)
QUANTA Flash Ro52 is a chemiluminescent immunoassay for the semi-quantitative determination of IgG anti-Ro52 autoantibodies in human serum. The presence of anti-Ro52 autoantibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of Systemic Lupus Erythematosus, Sjögren's Syndrome, Systemic Sclerosis, Idiopathic Inflammatory Myopathies.
QUANTA Flash Ro52 Calibrators are intended for use with the QUANTA Flash Ro52 Reagents for the determination of IgG anti-Ro52 autoantibodies in human serum. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash Ro52 Controls are intended for use with the QUANTA Flash Ro52 Reagents for quality control in the determination of IgG anti-Ro52 autoantibodies in human serum.
Substantial equivalence
The QUANTA Flash Ro52 Reagent, the QUANTA Flash Ro52 Calibrators and the QUANTA Flash Ro52 Controls have the same intended use and assay principle as the predicate device.
Comparison to predicate device
| Similarities | ||
|---|---|---|
| Item | QUANTA Flash Ro52 | Predicate Device |
| Intended use | Semi-quantitative determination of anti-Ro52 antibodies in human serum | Semi-quantitative detection of anti-Ro52 antibodies in human serum |
QUANTA Flash Ro52 reagent kit
8
| Similarities | ||
|---|---|---|
| Item | QUANTA Flash Ro52 | Predicate Device |
| Assay methodology | Solid phase (heterogeneous) | |
| immunoassay | Solid phase (heterogeneous) | |
| immunoassay | ||
| Antigen | Purified recombinant Ro52 antigen | Purified recombinant Ro52 antigen |
| Traceability | International Reference Preparation | |
| is not available | ||
| Results are traceable to in-house | ||
| Standards | International Reference Preparation is | |
| not available | ||
| Sample type | Serum | Serum |
| Shelf life | One year | One year |
| Differences | ||
|---|---|---|
| Item | QUANTA Flash Ro52 | Predicate Device |
| Detection/ | ||
| Operating principle | Chemiluminescent immunoassay | Enzyme-linked immunosorbent assay |
| Solid phase | Paramagnetic microparticles (beads) | 96-well plate |
| Conjugate | Isoluminol conjugated anti-human IgG | HRP conjugated anti-human IgG |
| Calibration | Lot specific Master Curve + two | |
| calibrators (sold separately) | SS-A 52 ELISA Low Positive | |
| (Included in the kit) |
QUANTA Flash Ro52 Calibrators
| Item | QUANTA Flash Ro52 Calibrators | Predicate Device |
|---|---|---|
| Intended use | QUANTA Flash Ro52 Calibrators are | |
| intended for use with the QUANTA | ||
| Flash Ro52 Reagents for the | ||
| determination of IgG anti-Ro52 | ||
| autoantibodies in human serum. Each | ||
| calibrator establishes a point of | ||
| reference for the working curve that | ||
| is used to calculate unit values. | No separate intended use; calibrator is | |
| part of the kit. | ||
| Analyte | Anti-Ro52 antibodies | Anti-Ro52 antibodies |
| Method | QUANTA Flash Ro52 | |
| chemiluminescent immunoassay | QUANTA Lite SS-A 52 ELISA | |
| Matrix | Human serum, buffer, protein | |
| stabilizer, and preservative | Human serum, buffer, protein | |
| stabilizer, and preservative | ||
| Unit | CU (Chemiluminescent units) | |
| (arbitrary) | units (arbitrary) |
9
| Item | QUANTA Flash Ro52 Calibrators | Predicate Device |
|---|---|---|
| Physico-chemical | ||
| characteristics | 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 Ro52 Controls
| Item | QUANTA Flash Ro52 Controls | Predicate Device |
|---|---|---|
| Intended use | QUANTA Flash Ro52 Controls are | |
| intended for use with the QUANTA | ||
| Flash Ro52 reagents for quality | ||
| control in the determination of IgG | ||
| anti-Ro52 autoantibodies in human | ||
| serum. | No separate intended use; controls are | |
| part of the kit. | ||
| Analyte | Anti-Ro52 antibodies | Anti-Ro52 antibodies |
| Method | QUANTA Flash Ro52 | |
| chemiluminescent immunoassay | QUANTA Lite SS-A 52 ELISA | |
| Matrix | Human serum, stabilizers, and | |
| preservative | Human serum with preservative | |
| Unit | CU (Chemiluminescent units) | |
| (arbitrary) | units (arbitrary) | |
| Physico-chemical | ||
| characteristics | Liquid, ready to use | Liquid, prediluted, ready to use |
| Levels | 2 (negative and positive) | 2 (ELISA negative and High positive) |
| Storage | 2-8 °C | 2-8 °C |
| Shelf life | One year | One year |
Analytical performance characteristics
Value assignment and traceability of Calibrators and Controls
There is currently no recognized international standard for the measurement of Ro52 antibodies. The Centers for Disease Control and Prevention ANA reference serum #2 (IS2073 - IIF ANA [speckled pattern]; anti-SS-B/La), #7 (IS2105 - Anti-SS-A/Ro), and #10 (IS2187 - Anti-Jo1) were tested for Ro52 and produced the following results: CDC ANA #2: 40.7 CU CDC ANA #7: 32.4 CU CDC ANA #10: 443.7 CU
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The QUANTA Flash Ro52 Calibrators and Controls are manufactured by diluting human serum that contains high titer of anti-Ro52 antibodies with buffer, containing protein stabilizer and 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.
Calibrator and Control values are directly traceable to in-house Standards that are used to create the Master Curves for the QUANTA Flash Ro52 assay.
| Material | Assigned Value |
|---|---|
| Ro52 Master Curve Standard 1 | 2.3 CU |
| Ro52 Master Curve Standard 2 | 10.1 CU |
| Ro52 Master Curve Standard 3 | 52.4 CU |
| Ro52 Master Curve Standard 4 | 98.0 CU |
| Ro52 Master Curve Standard 5 | 386.4 CU |
| Ro52 Master Curve Standard 6 | 1685.3 CU |
List of Ro52 Standards, Calibrators and Controls:
| Material | Manufacturing
Target Value | Manufacturing
Target Range |
|-----------------------|-------------------------------|-------------------------------|
| Ro52 Calibrator 1 | 10 CU | 8 – 12 CU |
| Ro52 Calibrator 2 | 400 CU | 360 – 440 CU |
| Ro52 Negative Control | 10 CU | 8 – 12 CU |
| Ro52 Positive Control | 50 CU | 40 – 60 CU |
Precision
The precision of the QUANTA Flash Ro52 assay was evaluated on 9 samples containing various concentrations of Ro52 antibodies in accordance with CLSI EP5-A2, Evaluation of Precision Performance of Quantitative Measurement Procedures - Approved Guideline: samples were run in duplicates, twice a day, for 21 days. Production reagent lot 131006 was used for the studies.
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: 1685.3 by further diluting it by 35 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 1685.3 CU, the highest value that can be reported is 58985 CU.
High concentration hook effect
To assess hook effect, measurement signal (relative light units, RLU) was examined for five high positive samples (results above the AMR) before and after automatic or manual dilution. All sera produced significantly higher RLU values (above the AMR) when used "as is" compared to the manually or automatically diluted ones (that were within the AMR), thereby confirming that high positive specimens above the analytical measuring range do not show hook effect up to 23005 CU in the Ro52 assay (the highest concentration that was tested).
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Linearity
The linearity of the AMR was evaluated by a study according to CLSI EP6-A, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approved Guideline. Production reagent lot 131006 was used for the study. Four serum samples with various Ro52 antibody concentrations were diluted in 10% increments (from 0% to 90% diluent) to obtain values that cover the AMR. The dilutions were assayed in duplicates. Percent recovery of obtained results was calculated compared to the expected results (based on the dilution factor). Moreover, obtained values were plotted against expected values, and linear regression analysis was performed. Only results within the AMR were included in the regression analysis.
Acceptance criteria:
- Recovery is between 80-120%, or ± 4 CU, whichever is greater.
- For linear regression analysis, slope is between 0.9-1.1, and R-is ≥ 0.95.
All four specimens showed dilution linearity individually.
| Sample | Test Range (CU) | Slope (95% CI) | R2 |
|---|---|---|---|
| 1 | 239.2 to 1618.2 | 0.97 (0.93 to 1.01) | 1.00 |
| 2 | 37.2 to 370.4 | 1.01 (0.99 to 1.04) | 1.00 |
| 3 | 11.0 to 104.2 | 1.02 (0.99 to 1.04) | 1.00 |
| 4 | 3.8 to 17.0 | 0.92 (0.86 to 0.98) | 0.99 |
The combined data yielded the following results with linear regression:
| Sample | Test Range (CU) | Slope (95% CI) | R2 |
|---|---|---|---|
| All | 3.8 to 1618.2 | 0.97 (0.96 to 0.97) | 1.00 |
Interference
The interference study was performed according to CLSI EPO7-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition. Three specimens were tested (negative: 16.5 CU; at the cutoff: 21.9 CU; positive: 64.9 CU). Interfering substances 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 Ro52 assay. Recovery of the unit values was calculated compared to control samples spiked with the same volume of diluents (10% of total). 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 10 mg/dL (recovery: 87% to 107%), hemoglobin up to 200 mg/dL (recovery: 89% to 107%), triglycerides up to 1000 mg/dL (recovery: 91% to 112%),
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cholesterol up to 224.3 mg/dL (recovery: 91% to 112%), and RF IgM up to 500 IU/mL (recovery: 92% to 108%).
Cross-reactivity
To test potential cross-reactivity with autoantibodies and infection-induced antibodies, results obtained on 199 of the total 233 control samples that were included in the clinical validation study were assessed. These samples were from patients with autoimmune diseases that are characterized with disease specific autoantibodies, or from patients with positive infectious disease serology. The composition of the cohort and the anti-Ro52 positivity rate is shown in the Table below:
| Diagnosis | Number of samples | # pos | % pos |
|---|---|---|---|
| Graves' Disease | 10 | 0 | 0.0% |
| Hashimoto Thyroiditis | 10 | 0 | 0.0% |
| Celiac Disease | 11 | 0 | 0.0% |
| Crohn's Disease | 20 | 0 | 0.0% |
| Ulcerative Colitis | 20 | 0 | 0.0% |
| HCV | 6 | 1 | 16.7% |
| HBV | 6 | 0 | 0.0% |
| CMV | 11 | 1 | 9.1% |
| EBV (with or without other infection) | 12 | 2 | 16.7% |
| HIV | 5 | 0 | 0.0% |
| Syphilis | 5 | 0 | 0.0% |
| Primary Antiphospholipid Syndrome | 15 | 0 | 0.0% |
| Vasculitis | 17 | 0 | 0.0% |
| Rheumatoid arthritis | 50 | 2 | 4.0% |
| Behçet's disease | 1 | 0 | 0.0% |
| Total controls | 199 | 6 | 3.0% |
Based on the results, the QUANTA Flash Ro52 assay does not show cross-reactivity with autoantibodies that are present in various autoimmune diseases, or antibodies against infectious agents.
Lot to lot comparison
Twenty unique samples and the Positive and Negative Controls (altogether 22 specimens) with various reactivity levels were tested in triplicates with three different reagent lots: 121005, 131006 and 141007. The samples covered the total analytical measuring range of the assay. Results were processed by linear
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510(k) Summary QUANTA Flash® Ro52
regression analysis and bias calculation according to CLSI EP09-A2, Method Comparison and Bias Calculation Using Patient Samples; Approved Guideline - Second Edition.
Pair-wise comparisons were performed between lot 121005 vs 131006, lot 121005 vs 141007 and lot 131006 vs 141007, considering individual replicates instead of the mean of replicates.
Acceptance criteria and results are in the Table below. All results were within the acceptance limits.
| | 121005 vs
131006 | 121005 vs
141007 | 131006 vs
141007 |
|------------------------------------------------------------------------------|---------------------|---------------------|---------------------|
| Acceptance criteria | | | |
| Weighted r: ≥0.975 for linear regression | 0.999 | 0.997 | 0.997 |
| Intercept of the regression line (constant bias):
± 15% of cut-off (3 CU) | 1.3 | 0.7 | -0.6 |
| Slope of the regression line (proportional bias): 0.9-
1.1 | 0.97 | 0.94 | 0.96 |
| Weighted S y/x: ≤ 0.5 | 0.043 | 0.06 | 0.06 |
| Predicted bias (difference) at cut-off: ±15% (3 CU) | 0.8 | -0.6 | -1.3 |
Stability
Shelf life
To establish the initial claim for shelf life, accelerated stability studies were performed for 4 weeks at 37°C ± 3°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 Ro52 to establish initial stability claim: the beads, the two Calibrators, and the negative and positive Controls. 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°C (control) to those stored at 37 ± 3°C (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:
With regression analysis, the lower 95% Cl interval of the regression line is ≥ 85% at 2 weeks, and no individual data point has ≤ 75% recovery at 2 weeks.
- Controls and Calibrators:
With regression analysis, the lower 95% Cl interval of the regression line is ≥ 90% at 2 weeks, and no individual data point has ≤ 80% recovery at 2 weeks.
Beads
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Testing was performed on three lots of Ro52 coupled beads using up to 6 characterized samples with various reactivity levels.
All three lots of beads retained > 85% reactivity (considering the 95% Cl) after two weeks at 37 ± 3°C, and therefore pass the acceptance criteria for one year expiration date.
Calibrators and Controls
Testing was performed on three lots of Ro52 Calibrators and Controls. All Calibrators and Controls maintained > 90% reactivity (considering the 95% CI) when sored at 37 ± 3°C for 2 weeks, and therefore pass the acceptance criteria for one year expiration dating.
In-use (onboard) stability
Calibrators
Onboard stability claim: 4 calibrations, or 8 hours onboard
During assessing 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.
A total of 5 successful calibrations were performed over an 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 (each vial run in duplicate) was used to establish baseline value, and then additional 19 runs (each vial run in singleton) were performed. 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 against the mean of duplicates of each vial from the first run. 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 115% at run 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
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To establish the in-use stability of the QUANTA Flash Ro52 reagent cartridge, three lots of cartridges were tested with up to 4 serum specimens (with different reactivity levels) along with the Negative and Positive Controls. The specimens were tested periodically up to 49 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.
The onboard stability results of the three lots are as follows:
RP0002: 36 days
121005: 36 days
141007: 42 days
Using these criteria, the in-use (onboard) stability of Ro52 reagent cartridge was set at 36 days.
Real time stability
Real time stability testing was performed at 3, 6, 9 and 12 and 18 months on the Calibrators and Controls, to support the one year expiration, and at 3, 6, 9 and 12 months on one lot of reagent cartridge, and at 0 and 19 months on another lot of reagent cartridge.
Each control was tested in triplicates at each time point.
- Acceptance criteria: results should fall within their acceptable ranges as it was established at the release of the controls.
Calibrators were tested in triplicates at each time point as it is done during calibration. Averages of the triplicates were compared to the value that was assigned to the Calibrators at release.
- Acceptance criteria: % recovery of the average of the triplicates is between 85% and 115%, and %CV of the triplicates is