(264 days)
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 derived from calibrators.
No
This device is an in vitro diagnostic (IVD) immunoassay designed for the semi-quantitative determination of antibodies to aid in the diagnosis of rheumatoid arthritis, not for treating or preventing disease.
Yes
The "Intended Use / Indications for Use" section explicitly states that the device is "an aid in the diagnosis of rheumatoid arthritis."
No
The device is a chemiluminescent immunoassay that runs on a fully automated closed system (BIO-FLASH instrument) which includes liquid handling hardware, a luminometer, and reagent cartridges. While it utilizes software, it is fundamentally a hardware-based in vitro diagnostic device.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is for the "semi-quantitative determination of IgG anti-CCP3 antibodies in human serum." It also states that the results are "an aid in the diagnosis of rheumatoid arthritis." This clearly indicates that the device is intended to be used in vitro (outside the body) to examine a human specimen (serum) for diagnostic purposes.
- Device Description: The description details a laboratory-based assay that uses reagents and an automated instrument (BIO-FLASH®) to analyze serum samples. This is consistent with the nature of an in vitro diagnostic device.
- Components: The kit includes reagents, calibrators, and controls, which are typical components of an IVD assay used for quantitative or semi-quantitative measurements.
- Performance Studies: The document describes clinical performance studies using human serum samples to determine sensitivity and specificity for diagnosing rheumatoid arthritis. This type of validation is required for IVD devices.
- Predicate Device: The mention of a "Predicate Device(s)" with a K number (K052264) indicates that this device is being compared to a previously cleared IVD device, which is a standard process for regulatory submission of new IVDs.
N/A
Intended Use / Indications for Use
QUANTA Flash® CCP3 is a chemiluminescent immunoassay for the semi-quantitative determination of IgG anti-CCP3 antibodies in human serum. The presence of anti-CCP3 antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis.
QUANTA Flash® CCP3 Calibrators are intended for use with the QUANTA Flash® CCP3 chemiluminescent immunoassay for the determination of IgG anti-CCP3 antibodies in human serum. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash® CCP3 Controls are intended for use with the QUANTA Flash® CCP3 chemiluminescent immunoassay for quality control in the determination of IgG anti-CCP3 antibodies in human serum.
Product codes
NHX, JIT, JJX
Device Description
The QUANTA Flash CCP3 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 CCP3 assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH instrument.
Synthetic cyclic citrullinated peptide 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 diluted 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 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 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-CCP3 antibodies bound to the corresponding beads.
For quantitation, the QUANTA Flash CCP3 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 CCP3 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 CCP3 kit contains the following materials:
One (1) QUANTA Flash CCP3 Reagent Cartridge
One (1) vial of Resuspension buffer
One (1) Transfer pipette
The QUANTA Flash CCP3 reagent cartridge contains the following reagents for 100 determinations:
- a. CCP3 coated paramagnetic beads, lyophilized.
- b. Assay buffer - colored pink, containing Tris-buffered saline, Tween 20, protein stabilizers and preservatives.
- C. Tracer IgG - Isoluminol labeled anti-human IgG antibodies in buffer, containing protein stabilizers and preservative.
The QUANTA Flash CCP3 Calibrators kit contains two vials of Calibrator 1 and two vials of Calibrator 2:
QUANTA Flash CCP3 Calibrators:
- QUANTA Flash CCP3 Calibrator 1: Two (2) barcode labeled tubes containing 0.7 mL prediluted, ready to use reagent. Calibrators contain human antibodies to CCP3 in stabilizer and preservative.
- QUANTA Flash CCP3 Calibrator 2: Two (2) barcode labeled tubes containing 0.7 mL prediluted, ready to use reagent. Calibrators contain human antibodies to CCP3 in stabilizer and preservative.
The QUANTA Flash CCP3 Controls kit contains two vials of Negative Control and two vials of Positive Control:
QUANTA Flash CCP3 Controls:
- QUANTA Flash CCP3 Negative Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to CCP3 in stabilizer and preservative.
- QUANTA Flash CCP3 Positive Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to CCP3 in stabilizer and preservative.
Mentions image processing
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Mentions AI, DNN, or ML
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Input Imaging Modality
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Anatomical Site
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Indicated Patient Age Range
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Intended User / Care Setting
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Description of the training set, sample size, data source, and annotation protocol
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Description of the test set, sample size, data source, and annotation protocol
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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical Performance Characteristics:
- Quantitation and units of measure: The assay measures chemiluminescent units (CU). Traceability to IUIS-CDC reference reagent for ACPA (P/N IS2723 L/N 08-0202) was established, with 100 U/mL correlating to 379.5 CU. The BIO-FLASH software converts CU to U/mL. The 20 CU cutoff equals 5.3 U/mL.
- Value assignment and traceability of Calibrators and Controls: Calibrators and Controls are manufactured by diluting human serum with high titer antibodies, tested for infectious substances. Target CU values are achieved through trial dilutions, bulk testing, and adjustment. Final value assignment is determined by testing on at least two instruments with at least two lots of reagent cartridge in 10 replicates. Values are traceable to in-house standards.
- Precision: Evaluated on 8 samples in accordance with CLSI EP05-A2, run in duplicates, twice a day, for 20 days (80 replicates per sample).
- Key results: Total %CV values ranged from 5.0% to 6.8%, which is within the acceptance criterion of 2776.8 CU after 20-fold dilution, bringing measured value within AMR. Highest reportable value is 55536 CU. No high concentration hook effect was observed up to 113191 CU.
- Linearity: Evaluated according to CLSI EPG-A. Five serum samples with various CCP3 antibody concentrations were diluted with negative serum in 10% increments and assayed in duplicates.
- Key results: Recovery was between 80-120% or ± 4 CU. Slope for linear regression was between 0.9-1.1 and R-squared was ≥ 0.95. All five specimens showed dilution linearity individually. The combined data for the range 6.6 to 2772.3 CU yielded a slope of 1.07 (1.05 to 1.10) and R2 of 0.99, demonstrating linearity of the analytical measuring range.
- Interference: Performed according to CLSI EPO7-A2. Three specimens were tested (negative, low positive, positive) with interfering substances spiked at three different concentrations in 10% of total specimen volume, assessed in triplicates.
- Key results: No interference detected with bilirubin up to 10 mg/dL (recovery: 101% to 102%), hemoglobin up to 200 mg/dL (recovery: 97% to 108%), triglycerides up to 1000 mg/dL (recovery: 98% to 111%), and cholesterol up to 224.3 mg/dL (recovery: 98% to 111%). Rheumatoid factor interference was not tested.
- Cross-reactivity: Tested on 234 control samples from patients with autoimmune diseases or positive infectious disease serology.
- Key results: Overall 2.1% positivity rate across various conditions (e.g., SLE 3.8%, Primary Biliary Cirrhosis 14.3%, AIH 5.3%, Celiac disease 5.0%). The assay does not show cross-reactivity with autoantibodies in various autoimmune diseases or with antibodies against infectious agents.
- Lot to lot comparison: Five unique samples (various reactivity levels) tested with three different reagent lots. Samples tested in replicates of 5, once per day, for 5 days according to CLSI EPO5-A3.
- Key results: Between lot %CV ranged from 2.8% to 4.0%, meeting acceptance criteria of
§ 866.5775 Rheumatoid factor immunological test system.
(a)
Identification. A rheumatoid factor immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the rheumatoid factor (antibodies to immunoglobulins) in serum, other body fluids, and tissues. Measurement of rheumatoid factor may aid in the diagnosis of rheumatoid arthritis.(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
October 23, 2015
INOVA DIAGNOSTICS, INC. DR. GABRIELLA LAKOS DIRECTOR, RESEARCH AND DEVELOPMENT 9900 OLD GROVE ROAD SAN DIEGO, CA 92131-1638
Re: K143754 Trade/Device Name: Quanta Flash® CCP3 Quanta Flash® CCP3 Calibrators Quanta Flash® CCP3 Controls Regulation Number: 21 CFR 866.5775 Regulation Name: Rheumatoid Factor Immunological Test System Regulatory Class: Class II Product Code: NHX, JIT, JIT, JJX Dated: August 19, 2015 Received: August 20, 2015
Dear Dr. Lakos:
This letter corrects our substantially equivalent letter of September 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
1
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 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 regulation (21 CFR Part 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" (21CFR 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 -S
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
2
Indications for Use
510(k) Number (if known) K143754
Device Name
QUANTA Flash® CCP3, QUANTA Flash® CCP3 Calibrators, QUANTA Flash® CCP3 Controls
Indications for Use (Describe)
QUANTA Flash CCP3 is a chemiluminescent immunoassay for the semi-quantitative determination of IgG anti-CCP3 antibodies in human serum. The presence of anti-CCP3 antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis.
QUANTA Flash CCP3 Calibrators are intended for use with the QUANTA Flash CCP3 chemiluminescent immunoassay for the determination of IgG anti-CCP3 antibodies in human serum. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash CCP3 Controls are intended for use with the QUANTA Flash CCP3 chemiluminescent immunoassay for quality control in the determination of IgG anti-CCP3 antibodies in human serum.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| ☒ Prescription Use (Part 21 CFR 801 Subpart D) | |
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary
QUANTA Flash® CCP3 QUANTA Flash® CCP3 Calibrators QUANTA Flash® CCP3 Controls
Table of Contents
| Administrative data…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… | |
|---|---|
| Predicate device | |
| Device description | |
| Intended use(s) | |
| Indications for use…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… | |
| Substantial equivalence | |
| Comparison to predicate device | |
| Analytical performance characteristics | |
| Quantitation and units of measure | |
| 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 |
4
Comparison with predicate device ............................................................................................................................................
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® CCP3
QUANTA Flash® CCP3 Calibrators
QUANTA Flash® CCP3 Controls |
| Scientific contact: | Gabriella Lakos, Director of Research, Rheumatology
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 |
| Device name (assay kit): | Proprietary name: QUANTA Flash® CCP3
Common name: Anti-CCP3 Chemiluminescent Immunoassay
Classification name: antibodies, anti-cyclic citrullinated peptide (ccp) |
| Regulation Description | Rheumatoid factor immunological test system |
| Regulation Medical Specialty | Immunology |
| Review Panel | Immunology |
| Product Code | NHX |
5
| Regulation Number | 866.5775 | |
|---|---|---|
| Device Class | 2 | |
| Device name (Calibrators): | Proprietary name: | |
| Common name: | ||
| Classification name: | QUANTA Flash® CCP3 Calibrators | |
| CCP3 Calibrators | ||
| 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: | |
| Common name: | ||
| Classification name: | QUANTA Flash® CCP3 Controls | |
| CCP3 Controls | ||
| 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
QUANTA Lite® CCP3 IgG ELISA, 510(k) number: K052264
Device description
The QUANTA Flash CCP3 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 CCP3 assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH instrument.
Synthetic cyclic citrullinated peptide is coated onto paramagnetic beads. The bead suspension is
6
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 diluted 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 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 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-CCP3 antibodies bound to the corresponding beads.
For quantitation, the QUANTA Flash CCP3 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 CCP3 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 CCP3 kit contains the following materials:
One (1) QUANTA Flash CCP3 Reagent Cartridge One (1) vial of Resuspension buffer One (1) Transfer pipette
The QUANTA Flash CCP3 reagent cartridge contains the following reagents for 100 determinations:
- a. CCP3 coated paramagnetic beads, lyophilized.
- b. Assay buffer - colored pink, containing Tris-buffered saline, Tween 20, protein stabilizers and preservatives.
- C. Tracer IgG - Isoluminol labeled anti-human IgG antibodies in buffer, containing protein stabilizers and preservative.
The QUANTA Flash CCP3 Calibrators kit contains two vials of Calibrator 1 and two vials of Calibrator 2:
QUANTA Flash CCP3 Calibrators:
- QUANTA Flash CCP3 Calibrator 1: Two (2) barcode labeled tubes containing 0.7 mL prediluted, ready to use reagent. Calibrators contain human antibodies to CCP3 in stabilizer
7
and preservative.
- QUANTA Flash CCP3 Calibrator 2: Two (2) barcode labeled tubes containing 0.7 mL । prediluted, ready to use reagent. Calibrators contain human antibodies to CCP3 in stabilizer and preservative.
The QUANTA Flash CCP3 Controls kit contains two vials of Negative Control and two vials of Positive Control:
QUANTA Flash CCP3 Controls:
- QUANTA Flash CCP3 Negative Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to CCP3 in stabilizer and preservative.
- । QUANTA Flash CCP3 Positive Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain human antibodies to CCP3 in stabilizer and preservative.
Intended use(s)
QUANTA Flash CCP3 is a chemiluminescent immunoassay for the semi-quantitative determination of IgG anti-CCP3 antibodies in human serum. The presence of antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis.
QUANTA Flash CCP3 Calibrators are intended for use with the QUANTA Flash CCP3 chemiluminescent immunoassay for the determination of IgG anti-CCP3 antibodies in human serum. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash CCP3 Controls are intended for use with the QUANTA Flash CCP3 chemiluminescent immunoassay for quality control in the determination of IgG antibodies in human serum.
Indications for use
Same as Intended use.
Substantial equivalence
The QUANTA Flash CCP3 Reagent, the QUANTA Flash CCP3 Calibrators and the QUANTA Flash CCP3 Controls have the same intended use and assay principle as the predicate device.
8
Comparison to predicate device
QUANTA Flash CCP3 reagent kit
| Similarities | ||
|---|---|---|
| Item | QUANTA Flash CCP3 | Predicate Device |
| Intended use | Semi-quantitative determination of anti-CCP3 antibodies in human serum | Semi-quantitative detection of anti-CCP3 antibodies in human serum, citrated and EDTA plasma |
| Assay methodology | Solid phase (heterogeneous) immunoassay | Solid phase (heterogeneous) immunoassay |
| Antigen | Synthetic peptide | Synthetic peptide |
| Shelf life | One year | One year |
| Differences | ||
|---|---|---|
| Item | QUANTA Flash CCP3 | Predicate Device |
| Detection/ | ||
| Operating principle | Chemiluminescent immunoassay | Enzyme-linked immunosorbent assay |
| Sample type | Serum | Serum; citrated or EDTA plasma |
| Solid phase | Paramagnetic microparticles (beads) | 96-well polystyrene plate |
| Conjugate | Isoluminol conjugated anti-human | |
| IgG | HRP conjugated anti-human IgG | |
| Calibration | Lot specific Master Curve + two | |
| calibrators (sold separately) | CCP3 ELISA Low Positive, and | |
| Calibrators A through E | ||
| (Included in the kit) |
QUANTA Flash CCP3 Calibrators
| Item | QUANTA Flash CCP3 Calibrators | Predicate Device |
|---|---|---|
| Intended use | For use with the QUANTA Flash CCP3 | |
| Reagents Each calibrator establishes | ||
| a point of reference for the working | ||
| curve that is used to calculate unit | ||
| values. | No separate intended use; calibrators | |
| are part of the kit. | ||
| Analyte | Anti-CCP3 antibodies | Anti-CCP3 antibodies |
| Method | QUANTA Flash CCP3 | |
| chemiluminescent immunoassay | QUANTA Lite CCP3 IgG ELISA | |
| Matrix | Human serum, stabilizer, and | |
| preservative | Human serum, buffer, protein | |
| stabilizer, and preservative | ||
| Unit | CU (Chemiluminescent units) | |
| (arbitrary) | ||
| U/mL (based on IUIS – CDC reference | ||
| reagent) | units (arbitrary) | |
| Physico-chemical | Liquid, prediluted, ready to use | Liquid, prediluted, ready to use |
9
| Item | QUANTA Flash CCP3 Calibrators | Predicate Device |
|---|---|---|
| characteristics | ||
| Storage | 2-8 °C | 2-8 °C |
| Shelf life | One year | One year |
QUANTA Flash CCP3 Controls
| Item | QUANTA Flash CCP3 Controls | Predicate Device |
|---|---|---|
| Intended use | For use with the QUANTA Flash CCP3 reagents for quality control in the determination of IgG anti-CCP3 autoantibodies in human serum. | No separate intended use; controls are part of the kit. |
| Analyte | Anti-CCP3 antibodies | Anti-CCP3 antibodies |
| Method | QUANTA Flash CCP3 chemiluminescent immunoassay | QUANTA Lite CCP3 IgG ELISA |
| Matrix | Human serum, stabilizers, and preservative | Human serum, buffer, protein stabilizer, and preservative |
| Unit | CU (Chemiluminescent units) (arbitrary) | |
| U/mL (based on IUIS – CDC reference reagent) | 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
Quantitation and units of measure
For quantitation, the QUANTA Flash CCP3 assay utilizes a lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. The Master Curve for QUANTA Flash CCP3 consists of 7 Standards. These Master Curve Standards are used to create the lot specific Master Curve during the manufacturing procedure.
List of CCP3 Standards:
| Material | Assigned Value |
|---|---|
| CCP3 Master Curve Standard 1 | 4.6 CU |
| CCP3 Master Curve Standard 2 | 14.6 CU |
| CCP3 Master Curve Standard 3 | 42.2 CU |
| CCP3 Master Curve Standard 4 | 124.3 CU |
| CCP3 Master Curve Standard 5 | 313.7 CU |
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| CCP3 Master Curve Standard 6 | 996.1 CU |
|---|---|
| CCP3 Master Curve Standard 7 | 2776.8 CU |
To establish traceability to a reference material, the IUIS-Centers for Disease Control and Prevention (CDC) reference reagent for anti-citrullinated peptide/protein antibodies (ACPA) (P/N IS2723 L/N 08-0202) was tested, and the value of 379.5 CU was obtained. The reference reagent has an assigned value of 100 U/mL.
Using the 100 U/mL assigned value of the reference material, the correlation between CU and U/mL values was established. Based on the obtained correlation, the BIO-FLASH software is able to automatically convert anti-CCP3 CU values into U/mL values. The 20 CU cutoff value equals 5.3 U/mL.
Value assignment and traceability of Calibrators and Controls
The QUANTA Flash CCP3 Calibrators and Controls are manufactured by diluting human serum that contains high titer of antibodies with 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 the in-house Standards that are used to create the Master Curves for the QUANTA Flash CCP3 assay.
| Material | Manufacturing
Target Value | Manufacturing
Target Range |
|-----------------------|-------------------------------|-------------------------------|
| CCP3 Calibrator 1 | 14 CU | 10-16 CU |
| CCP3 Calibrator 2 | 300 CU | 270 - 330 CU |
| CCP3 Negative Control | 10 CU | 8-12 CU |
| CCP3 Positive Control | 50 CU | 40-60 CU |
CCP3 Calibrators and Controls with target manufacturing values:
Precision
The precision of the QUANTA Flash CCP3 assay was evaluated on 8 samples containing various concentrations of CCP3 antibodies in accordance with CLSI EP05-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.
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| QUANTA Flash CCP3 | Within Run | Between Runs | Between Days | Total | ||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Sample ID | Number of replicates | Mean (CU) | SD (CU) | CV (%) | SD (CU) | CV (%) | SD (CU) | CV (%) | SD (CU) | CV (%) |
| 1 | 80 | 11.7 | 0.7 | 5.8 | 0.0 | 0.0 | 0.4 | 3.5 | 0.8 | 6.8 |
| 2 | 80 | 21.3 | 1.0 | 4.6 | 0.7 | 3.2 | 0.7 | 3.5 | 1.4 | 6.6 |
| 3 | 80 | 21.4 | 0.8 | 3.6 | 0.6 | 2.8 | 0.9 | 4.1 | 1.3 | 6.1 |
| 4 | 80 | 23.0 | 1.0 | 4.4 | 0.0 | 0.0 | 0.6 | 2.8 | 1.2 | 5.2 |
| 5 | 80 | 55.9 | 1.4 | 2.6 | 1.1 | 1.9 | 2.1 | 3.8 | 2.8 | 5.0 |
| 6 | 80 | 195.1 | 8.8 | 4.5 | 6.0 | 3.1 | 1.3 | 0.6 | 10.7 | 5.5 |
| 7 | 80 | 1155.3 | 56.2 | 4.9 | 20.3 | 1.8 | 32.6 | 2.8 | 68.0 | 5.9 |
| 8 | 80 | 2210.3 | 119.9 | 5.4 | 42.3 | 1.9 | 36.6 | 1.7 | 132.3 | 6.0 |
Acceptance criteria: Total %CV: 2776.8 after further diluting it by 20 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 directly measured is 2776.8 CU, the highest value that can be reported is 55536 CU.
High concentration hook effect
To assess hook effect, measurement signal (relative light units, RLU) was examined for four 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 113191 CU in the CCP3 assay (the highest concentration that was tested).
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 with various CCP3 antibody concentrations were diluted with negative serum in 10% increments (from
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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 calculated compared to the expected mean results (based on the dilution factor). Moreover, obtained values were plotted against expected values, and linear regression analysis was performed.
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 five specimens showed dilution linearity individually.
| Sample | Test Range (CU) | Slope (95% CI) | R2 |
|---|---|---|---|
| 1 | 358.3 to 2772.3 | 1.00 (1.00 to 1.00) | 1.00 |
| 2 | 190.7 to 2030.0 | 0.99 (0.96 to 1.03) | 0.99 |
| 3 | 82.5 to 705.4 | 1.00 (0.95 to 1.04) | 0.99 |
| 4 | 14.0 to 134.5 | 0.94 (0.88 to 1.00) | 0.98 |
| 5 | 6.6 to 34.9 | 0.97 (0.93 to 1.02) | 0.99 |
The combined data yielded the following results with linear regression:
| Sample | Test Range (CU) | Slope (95% CI) | D2 |
|---|---|---|---|
| All | 6.6 to 2772.3 | 1.07 (1.05 to 1.10) | 0.99 |
These data demonstrate the linearity of the analytical measuring range (4.6 CU - 2776.8 CU) of the QUANTA Flash CCP3 assay.
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: 10.1 CU; low positive: 40.5 CU; positive: 939.8 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 CCP3 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: 101% to 102%), hemoglobin up to 200 mg/dL (recovery: 97% to 108%), triglycerides up to 1000 mg/dL (recovery: 98% to 111%), and cholesterol up to 224.3 mg/dL (recovery: 98% to 11%). Interference of rheumatoid factor was not tested.
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Cross-reactivity
To test potential cross-reactivity with autoantibodies and infection-induced antibodies, results obtained on 234 of the total 376 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-CCP3 positivity rate is shown in the Table below:
| Diagnosis | Number of samples | # pos | % pos |
|---|---|---|---|
| Systemic Lupus Erythematosus | 53 | 2 | 3.8% |
| HBV | 22 | 0 | 0.0% |
| HCV | 13 | 0 | 0.0% |
| Limited Systemic Sclerosis | 8 | 0 | 0.0% |
| Mixed Connective Tissue Disease | 7 | 0 | 0.0% |
| Primary Biliary Cirrhosis | 7 | 1 | 14.3% |
| Vasculitis | 6 | 0 | 0.0% |
| Wegener's Granulomatosis | 1 | 0 | 0.0% |
| Parvo virus infection | 12 | 0 | 0.0% |
| Salmonella infection | 10 | 0 | 0.0% |
| Lyme disease | 25 | 0 | 0.0% |
| Ulcerative colitis (UC) | 11 | 0 | 0.0% |
| Autoimmune hepatitis (AIH) | 19 | 1 | 5.3% |
| Celiac disease (CD) | 20 | 1 | 5.0% |
| Sjögren's syndrome | 20 | 0 | 0.0% |
| Total | 234 | 5 | 2.1% |
Based on the results, the QUANTA Flash CCP3 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
Five unique samples with various reactivity levels were tested with three different reagent lots: 141004, 141005 and 151006. The samples covered the analytical measuring range of the assay. Samples were tested in replicates of 5, once per day, for 5 days according to CLSI EPO5-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples; Approved Guideline.
Data were analyzed with the Analyse-it for Excel method evaluation software to calculate between lot precision.
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| QUANTA Flash CCP3 | Between Lot Precision | |||
|---|---|---|---|---|
| Sample ID | Mean (CU) | N | SD (CU) | CV (%) |
| 1 | 16.2 | 75 | 0.6 | 3.9 |
| 2 | 24.9 | 75 | 0.7 | 2.8 |
| 3 | 77.5 | 75 | 2.6 | 3.4 |
| 4 | 341.1 | 75 | 10.2 | 3.0 |
| 5 | 1528.7 | 75 | 61.2 | 4.0 |
Acceptance criteria: Between lot %CV: