(90 days)
Not Found
No
The device description details a standard automated immunoassay system that uses predefined master and working curves for calculation, which is not indicative of AI/ML. There is no mention of AI, ML, or related terms in the document.
No
This device is an immunoassay designed for diagnostic purposes (detection of rheumatoid factor antibodies to aid in the diagnosis of rheumatoid arthritis), not for treating any condition.
Yes
The "Intended Use / Indications for Use" states that the device is "an aid in the diagnosis of rheumatoid arthritis (RA)", which explicitly indicates it is a diagnostic device.
No
The device description explicitly states that the assays are designed to run on the BIO-FLASH® instrument, which is described as a fully automated closed system with liquid handling hardware, luminometer, and a computer with software-user interface. The device also includes reagent cartridges containing physical reagents. This indicates the device is a system that includes both hardware and software components, 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 that the device is for the "quantitative determination of IgM rheumatoid factor (RF) antibodies in human serum" and the "semi-quantitative determination of IgA rheumatoid factor (RF) antibodies in human serum." It also states that the results are an "aid in the diagnosis of rheumatoid arthritis (RA)." 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 "chemiluminescent immunoassay" that runs on an automated instrument and uses reagents to detect antibodies in a patient serum sample. This is a typical description of an in vitro diagnostic test.
- Specimen Type: The device uses "human serum," which is a biological specimen.
- Diagnostic Purpose: The results are used as an "aid in the diagnosis of rheumatoid arthritis (RA)."
All of these points align with the definition of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
QUANTA Flash RF IgM is a chemiluminescent immunoassay for the quantitative determination of IgM rheumatoid factor (RF) antibodies in human serum. The presence of IgM RF antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis (RA).
QUANTA Flash RF IgA is a chemiluminescent immunoassay for the semi-quantitative determination of lgA rheumatoid factor (RF) antibodies in human serum. The presence of IgA RF antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis (RA).
Product codes (comma separated list FDA assigned to the subject device)
DHR
Device Description
The principle of the assays is chemiluminescent microparticle immunoassay, a variation of solid phase immunoassay. The QUANTA Flash® RF IgM and QUANTA Flash® RF IgA assays are 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® RF IgM and QUANTA Flash® RF IgA assays utilize a reagent cartridge format, which is compatible with the BIO-FLASH® instrument.
Rabbit polyclonal antibodies are coated onto paramagnetic beads, which are stored in the reagent cartridge under conditions that preserve the antibody in its reactive state. When the assay cartridge is ready to be used for the first time, the entire cartridge is inverted several times to thoroughly mix the reagents. The reagent cartridge is then loaded onto the BIO-FLASH instrument.
A patient serum sample is diluted 1:22.7 by the instrument using system rinse in a disposable plastic cuvette. An aliquot of the diluted patient serum, coupled beads, and assay buffer are combined into a second cuvette, and mixed. This cuvette is incubated at 37°C. The beads are then magnetized and washed several times. Isoluminol conjugated anti-human IgM (QUANTA Flash® RF IgM) or anti-human lgA (QUANTA Flash® RF IgA) antibody is then added to the cuvette, and incubated at 37°C. Again, the beads are magnetized and washed repeatedly. The isoluminol conjugate produces a luminescent reaction when "Trigger" reagents are added to the light produced from this reaction is measured as Relative Light Units (RLU) by the BIO-FLASH optical system. RLU values are proportional to the amount of bound isoluminol conjugate, which in turn is proportional to the amount of RF antibodies bound to the antibodies on the beads.
The QUANTA Flash RF IgM and QUANTA Flash RF IgA assays utilize a predefined lot specific Master Curve that is uploaded into the instrument through the reagent cartridge barcode. Based on the results obtained by running the Calibrators, an instrument specific Working Curve is created, which is used by the software to calculate international units per milliliter (IU/mL) (QUANTA Flash® RF IgM) or chemiluminescent units (CU) (QUANTA Flash® RF IgA) from the RLU value obtained for each sample.
QUANTA Flash RF IgM Calibrators, QUANTA Flash RF IgM Controls, QUANTA Flash RF IgA Calibrators and QUANTA Flash RF IgA Controls are sold separately.
The QUANTA Flash® RF IgM Reagents / QUANTA Flash® RF IgA Reagents kit contains the following materials:
One (1) QUANTA Flash RF IgM / RF IgA Reagent Cartridge
QUANTA Flash RF IgM Reagent Cartridge contains the following reagents for 100 determinations:
- a. Rabbit pAb coated paramagnetic beads.
- b. Assay buffer - colored pink, containing protein stabilizers and preservatives.
- Tracer IgM Isoluminol labeled anti-human IgM antibody, containing buffer, protein C. stabilizers and preservative.
QUANTA Flash RF IgA Reagent Cartridge contains the following reagents for 100 determinations:
- a. Rabbit pAb coated paramagnetic beads.
- b. Assay buffer - colored pink, containing protein stabilizers and preservatives.
- Tracer IgA Isoluminol labeled anti-human IgA antibody, containing buffer, protein C. stabilizers and preservative.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Human serum
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 RF IgM and the QUANTA Flash RF IgA assays. A total of 706 characterized samples were included in this Validation Set. All samples were run on the QUANTA Flash RF IgM and the QUANTA Flash RF IgA assays. The distribution of the cohort and the RF positivity rate is in the Table below:
| Patient Group | N | RF IgM
N Positive | RF IgM
% Positive | RF IgA
N Positive | RF IgA
% Positive |
|---------------------------------|-----|----------------------|----------------------|----------------------|----------------------|
| Ulcerative Colitis | 50 | 8 | 16.0% | 10 | 20.0% |
| Crohn's Disease | 30 | 3 | 10.0% | 2 | 6.7% |
| Osteoarthritis | 30 | 3 | 10.0% | 0 | 0.0% |
| Autoimmune Hepatitis | 30 | 0 | 0.0% | 1 | 3.3% |
| Sjögren's Syndrome | 30 | 7 | 23.3% | 6 | 20.0% |
| Systemic Sclerosis | 30 | 9 | 30.0% | 10 | 33.3% |
| Celiac Disease | 29 | 0 | 0.0% | 0 | 0.0% |
| Systemic Lupus Erythematosus | 28 | 4 | 14.3% | 4 | 14.3% |
| Hepatitis B Virus | 20 | 1 | 5.0% | 0 | 0.0% |
| Hepatitis C Virus | 19 | 0 | 0.0% | 1 | 5.3% |
| Polymyalgia Rheumatica | 15 | 2 | 13.3% | 0 | 0.0% |
| Psoriatic Arthritis | 12 | 0 | 0.0% | 0 | 0.0% |
| Syphilis | 12 | 1 | 8.3% | 0 | 0.0% |
| Parvo virus infection | 10 | 2 | 20.0% | 2 | 20.0% |
| Lyme disease | 10 | 0 | 0.0% | 0 | 0.0% |
| Fibromyalgia | 10 | 1 | 10.0% | 0 | 0.0% |
| Ankylosing Spondylitis | 8 | 0 | 0.0% | 0 | 0.0% |
| Mixed connective tissue disease | 8 | 1 | 12.5% | 1 | 12.5% |
| Hashimoto's disease | 8 | 0 | 0.0% | 0 | 0.0% |
| Grave's disease | 6 | 1 | 16.7% | 0 | 0.0% |
| Polymyositis | 10 | 3 | 30.0% | 2 | 20.0% |
| Dermatomyositis | 5 | 2 | 40.0% | 0 | 0.0% |
| Total Controls | 410 | 48 | 11.7% | 39 | 9.5% |
| Rheumatoid Arthritis | 296 | 206 | 69.6% | 168 | 56.8% |
| Total | 706 | - | - | - | - |
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Clinical sensitivity and specificity of the QUANTA Flash RF IgM were analyzed:
Sample size (N) = 706
Sensitivity: 69.6% (95% CI: 64.1 – 74.6%)
Specificity: 88.3% (95% CI: 84.8 - 91.1%)
Clinical sensitivity and specificity of the QUANTA Flash RF IgA were analyzed:
Sample size (N) = 706
Sensitivity: 56.8% (95% CI: 51.1 – 62.3%)
Specificity: 90.5% (95% CI: 87.3 – 93.0%)
Method comparison of the QUANTA Flash RF IgM with the predicate device using samples within the AMR (N=577):
NPA: 96.4 (95% CI: 93.6-98.0)
PPA: 81.1 (95% CI: 76.0 - 85.3)
TPA: 89.1 (95% CI: 86.3 - 91.4)
Spearman's rs of 0.85 (95% Cl, 0.82 - 0.87)
Method comparison of the QUANTA Flash RF IgA with the predicate device using samples within the AMR (N=612):
NPA: 97.5 (95% CI: 95.4–98.6)
PPA: 87.6 (95% CI: 82.5 – 91.3)
TPA: 94.0 (95% CI: 91.8 – 95.6)
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
QUANTA Flash RF IgM:
Sensitivity: 69.6%
Specificity: 88.3%
Positive Percent Agreement (PPA): 81.1
Negative Percent Agreement (NPA): 96.4
Total Percent Agreement (TPA): 89.1
QUANTA Flash RF IgA:
Sensitivity: 56.8%
Specificity: 90.5%
Positive Percent Agreement (PPA): 87.6
Negative Percent Agreement (NPA): 97.5
Total Percent Agreement (TPA): 94.0
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
QUANTA Lite® RF IgM ELISA, 510(k) number: K971614, QUANTA Lite® RF IgA ELISA, 510(k) number: K983084
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 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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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
April 17, 2019
Inova Diagnostics, Inc. Roger Albesa Manager, Research and Development 9900 Old Grove Rd. San Diego, California 92131-1638
Re: K190088
Trade/Device Name: QUANTA Flash RF IgM Reagents, QUANTA Flash RF IgA Reagents Regulation Number: 21 CFR 866.5775 Regulation Name: Rheumatoid factor immunological test system Regulatory Class: Class II Product Code: DHR Dated: January 16, 2019 Received: January 17, 2019
Dear Roger Albesa:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's
1
requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely.
Doug Jeffery Acting Deputy 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) Summary
QUANTA Flash® RF IgM Reagents QUANTA Flash® RF IgA Reagents
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 ............................................................................................................................................ Precision .................................................................................................................................................................... Reproducibility Studies ...................................................................................................................................................... Limit of Blank (LoB), Limit of Detection (LoD), and Limit of Quantitation (LoQ)............................................................................................... Analytical Measuring Range (AMR) ............................................................................................................................................. Auto-rerun function and reportable results ................................................................................................................................... High concentration hook effect................................................................................................................................................ Linearity .................................................................................................................................................................... Interference ................................................................................................................................................................. Sample Stability and Handling ................................................................................................................................................ Reagent Stability ............................................................................................................................................................ Cut-off, reference range...................................................................................................................................................... Clinical performance characteristics.......................................................................................................................................... Clinical sensitivity, specificity ............................................................................................................................................ Expected values............................................................................................................................................................... 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.
4
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® RF IgM Reagents
QUANTA Flash® RF IgA Reagents | |
| Scientific contact: | Roger Albesa, Manager, Research and Development
Inova Diagnostics, Inc.
9900 Old Grove Road, San Diego, CA, 92131
Phone: 858-586-9900 x1391
Fax: 858-863-0025
Email: ralbesa@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 x1381
Fax: 858-863-0025
Email: relliot@inovadx.com | |
| Device name (assay kit): | Proprietary name: | QUANTA Flash® RF IgM Reagents
QUANTA Flash® RF IgA Reagents |
| | Common name: | Rheumatoid factor IgM, IgA Chemiluminescence
Immunoassay |
| | Classification name: | System, Test, Rheumatoid Factor |
| Regulation Description | Rheumatoid Factor immunological test system | |
| Regulation Medical Specialty | Immunology | |
| Review Panel | Immunology | |
| Product Code | DHR | |
| Regulation Number | 866.5775 | |
| Device Class | 2 | |
5
Predicate device
QUANTA Lite® RF IgM ELISA, 510(k) number: K971614. Date declared: April 30, 1997. QUANTA Lite® RF IgA ELISA, 510(k) number: K983084. Date declared: September 2, 1998.
Device description
The principle of the assays is chemiluminescent microparticle immunoassay, a variation of solid phase immunoassay. The QUANTA Flash® RF IgM and QUANTA Flash® RF IgA assays are 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® RF IgM and QUANTA Flash® RF IgA assays utilize a reagent cartridge format, which is compatible with the BIO-FLASH® instrument.
Rabbit polyclonal antibodies are coated onto paramagnetic beads, which are stored in the reagent cartridge under conditions that preserve the antibody in its reactive state. When the assay cartridge is ready to be used for the first time, the entire cartridge is inverted several times to thoroughly mix the reagents. The reagent cartridge is then loaded onto the BIO-FLASH instrument.
A patient serum sample is diluted 1:22.7 by the instrument using system rinse in a disposable plastic cuvette. An aliquot of the diluted patient serum, coupled beads, and assay buffer are combined into a second cuvette, and mixed. This cuvette is incubated at 37°C. The beads are then magnetized and washed several times. Isoluminol conjugated anti-human IgM (QUANTA Flash® RF IgM) or anti-human lgA (QUANTA Flash® RF IgA) antibody is then added to the cuvette, and incubated at 37°C. Again, the beads are magnetized and washed repeatedly. The isoluminol conjugate produces a luminescent reaction when "Trigger" reagents are added to the light produced from this reaction is measured as Relative Light Units (RLU) by the BIO-FLASH optical system. RLU values are proportional to the amount of bound isoluminol conjugate, which in turn is proportional to the amount of RF antibodies bound to the antibodies on the beads.
The QUANTA Flash RF IgM and QUANTA Flash RF IgA assays utilize a predefined lot specific Master Curve that is uploaded into the instrument through the reagent cartridge barcode. Based on the results obtained by running the Calibrators, an instrument specific Working Curve is created, which is used by the software to calculate international units per milliliter (IU/mL) (QUANTA Flash® RF IgM) or chemiluminescent units (CU) (QUANTA Flash® RF IgA) from the RLU value obtained for each sample.
QUANTA Flash RF IgM Calibrators, QUANTA Flash RF IgM Controls, QUANTA Flash RF IgA Calibrators and QUANTA Flash RF IgA Controls are sold separately.
The QUANTA Flash® RF IgM Reagents / QUANTA Flash® RF IgA Reagents kit contains the following materials:
One (1) QUANTA Flash RF IgM / RF IgA Reagent Cartridge
6
QUANTA Flash RF IgM Reagent Cartridge contains the following reagents for 100 determinations:
- a. Rabbit pAb coated paramagnetic beads.
- b. Assay buffer - colored pink, containing protein stabilizers and preservatives.
- Tracer IgM Isoluminol labeled anti-human IgM antibody, containing buffer, protein C. stabilizers and preservative.
QUANTA Flash RF IgA Reagent Cartridge contains the following reagents for 100 determinations:
- a. Rabbit pAb coated paramagnetic beads.
- b. Assay buffer - colored pink, containing protein stabilizers and preservatives.
- Tracer IgA Isoluminol labeled anti-human IgA antibody, containing buffer, protein C. stabilizers and preservative.
Intended use(s)
QUANTA Flash RF IgM is a chemiluminescent immunoassay for the quantitative determination of IgM rheumatoid factor (RF) antibodies in human serum. The presence of IgM RF antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis (RA).
QUANTA Flash RF IgA is a chemiluminescent immunoassay for the semi-quantitative determination of lgA rheumatoid factor (RF) antibodies in human serum. The presence of IgA RF antibodies, in conjunction with clinical findings and other laboratory tests, is an aid in the diagnosis of rheumatoid arthritis (RA).
Indications for use
Same as Intended use.
Substantial equivalence
The QUANTA Flash RF IgM Reagents and the QUANTA Flash RF IgA Reagents have the same intended use and assay principle as the predicate device.
7
Comparison to predicate device
QUANTA Flash RF IgM Reagents
| Similarities | ||
|---|---|---|
| Item | QUANTA Flash RF IgM Reagents | QUANTA Lite RF IgM ELISA |
| Intended use | QUANTA Flash RF IgM is a | |
| chemiluminescent immunoassay for the | ||
| quantitative determination of IgM | ||
| rheumatoid factor (RF) antibodies in | ||
| human serum. The presence of IgM RF | ||
| antibodies, in conjunction with clinical | ||
| findings and other laboratory tests, is an | ||
| aid in the diagnosis of rheumatoid | ||
| arthritis (RA). | QUANTA Lite RF IgM is an enzyme- | |
| linked immunosorbent assay (ELISA) for | ||
| the semi-quantitative detection of IgM | ||
| rheumatoid factor (RF) antibodies in | ||
| patient sera. The presence of these | ||
| antibodies, when considered in | ||
| conjunction with other laboratory and | ||
| clinical findings, is an aid in the | ||
| diagnosis of rheumatoid arthritis (RA). | ||
| Assay methodology | Solid phase (heterogeneous) | |
| immunoassay | Solid phase (heterogeneous) | |
| immunoassay | ||
| Antigen | Rabbit polyclonal antibody | Rabbit polyclonal antibody |
| Sample type | Human serum | Human serum |
| Differences | ||
|---|---|---|
| Item | QUANTA Flash RF IgM Reagents | QUANTA Lite RF IgM ELISA |
| Detection/ | ||
| Operating principle | Chemiluminescent immunoassay | Enzyme-linked immunosorbent assay |
| Solid phase | Paramagnetic microparticles (beads) | 96-well polystyrene plate |
| Conjugate | Isoluminol conjugated monoclonal anti- | |
| human IgM antibody | HRP conjugated monoclonal anti- | |
| human IgM antibody | ||
| Units | International Units per milliliter (IU/mL) | Units (U) |
| Cut-off | 5.0 IU/mL | 6.0 Units |
| Analytical | ||
| Measuring Range | 0.3 - 490.0 IU/mL | 0.0 – 100.0 Units |
| Calibration | Lot specific Master Curve + Two | |
| calibrators (sold separately) | Five lot specific calibrators |
8
| Similarities | ||
|---|---|---|
| Item | QUANTA Flash RF IgA Reagents | QUANTA Lite RF IgA ELISA |
| Intended use | QUANTA Flash RF IgA is a | |
| chemiluminescent immunoassay for the | ||
| semi-quantitative determination of IgA | ||
| rheumatoid factor (RF) antibodies in | ||
| human serum. The presence of IgA RF | ||
| antibodies, in conjunction with clinical | ||
| findings and other laboratory tests, is an | ||
| aid in the diagnosis of rheumatoid | ||
| arthritis (RA). | QUANTA Lite RF IgA is an enzyme- | |
| linked immunosorbent assay (ELISA) for | ||
| the semi-quantitative detection of IgA | ||
| rheumatoid factor (RF) antibodies in | ||
| patient sera. The presence of these | ||
| antibodies, when considered in | ||
| conjunction with other laboratory and | ||
| clinical findings, is an aid in the | ||
| diagnosis of rheumatoid arthritis (RA). | ||
| Assay methodology | Solid phase (heterogeneous) | |
| immunoassay | Solid phase (heterogeneous) | |
| immunoassay | ||
| Antigen | Rabbit polyclonal antibody | Rabbit polyclonal antibody |
| Sample type | Human serum | Human serum |
QUANTA Flash RF IgA Reagents
| Differences | ||
|---|---|---|
| Item | QUANTA Flash RF IgA Reagents | QUANTA Lite RF IgA ELISA |
| Detection/ | ||
| Operating principle | Chemiluminescent immunoassay | Enzyme-linked immunosorbent assay |
| Solid phase | Paramagnetic microparticles (beads) | 96-well polystyrene plate |
| Conjugate | Isoluminol conjugated monoclonal anti- | |
| human IgA antibody | HRP conjugated monoclonal anti- | |
| human IgA antibody | ||
| Units | Chemiluminescent Units (CU) | Units (U) |
| Cut-off | 20.0 CU | 6.0 Units |
| Analytical | ||
| Measuring Range | 1.3 – 900.0 CU | 0.0 – 100.0 Units |
| Calibration | Lot specific Master Curve + Two | |
| calibrators (sold separately) | Five lot specific calibrators |
9
Analytical performance characteristics
Quantitation and units of measure
For quantitation, the QUANTA Flash RF IgM and QUANTA Flash RF IgA assays utilize a lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. The Master Curve for QUANTA Flash RF IgM and QUANTA Flash RF IgA consist of 6 different Standards each. These Master Curve Standards are used to create the lot specific Master Curve during the manufacturing procedure.
List of QUANTA Flash RF IgM Standards:
| Material | Assigned Value |
|---|---|
| RF IgM Master Curve Standard 1 | 0.0 IU/mL |
| RF IgM Master Curve Standard 2 | 1.9 IU/mL |
| RF IgM Master Curve Standard 3 | 7.8 IU/mL |
| RF IgM Master Curve Standard 4 | 31.0 IU/mL |
| RF IgM Master Curve Standard 5 | 124.1 IU/mL |
| RF IgM Master Curve Standard 6 | 496.5 IU/mL |
List of QUANTA Flash RF IgA Standards:
| Material | Assigned Value |
|---|---|
| RF IgA Master Curve Standard 1 | 0.0 CU |
| RF IgA Master Curve Standard 2 | 7.2 CU |
| RF IgA Master Curve Standard 3 | 14.4 CU |
| RF IgA Master Curve Standard 4 | 57.5 CU |
| RF IgA Master Curve Standard 5 | 230.0 CU |
| RF IgA Master Curve Standard 6 | 919.9 CU |
Precision
The precision of the QUANTA Flash RF IgM and QUANTA Flash RF IgA assays was evaluated on 10 samples for the IgM subtype and 9 samples for the IgA one, containing various concentrations of RF antibodies in accordance with CLSI EP05-A3, Evaluation of Quantitative Measurement Procedures; 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 repeatability (withinrun), between run, between day and within-laboratory precision) were calculated. Acceptance criteria: Total %CV: 490.0 IU/mL, for the QUANTA Flash RF IgM, or >900.0 CU, for the QUANTA Flash RF IgA, 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.
For the QUANTA Flash RF IgM, the highest value that can be directly measured is 490.0 IU/mL, so the highest value that can be reported is 9,800.0 IU/mL.
For the QUANTA Flash RF IgA, the highest value that can be directly measured is 900.0 CU, so the highest value that can be reported is 18,000.0 CU.
High concentration hook effect
To assess hook effect, measurement signal in relative light units (RLU) was examined by performing serial dilutions of two high positive samples (with results above the AMR of each assay when tested as neat samples). RLU values showed increasing antibody concentrations above the AMR for both assays, thereby confirming that high positive specimens above the AMR do not show hook effect up to 1,830.6 IU/mL for the QUANTA Flash RF IgM assay and 42,710.4 CU for the QUANTA Flash RF IgA assay (theoretical value calculated using the highest value in the AMR and its dilution factor).
Linearity
The linearity of the AMR of the QUANTA Flash RF IgM and QUANTA Flash RF IgA was evaluated by a study according to CLSI EP06-A, Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline. The linearity was evaluated using four human serum samples for the QUANTA Flash RF IgM and three human serum samples for the QUANTA Flash RF IgA with various RF antibody concentrations which were combined with another human serum sample containing low levels of RF antibodies in 10% increments (from 0% to 90% of low sample) to obtain values that cover the entire AMR of the assays. The dilutions were assayed in duplicates. Results were analyzed according to the guideline performing regression analysis and identifying the best fitting polynomial.
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Acceptance criteria:
- Best fitting polynomial is a linear one, otherwise, the difference between the best-fitting nonlinear and linear polynomial is less than 15% or ±0.75 IU/mL / ±3 CU for negative samples (allowable nonlinearity).
For the QUANTA Flash RF IgM, all samples have been found that the best fitting polynomial is a linear one except Sample 3, where the best fitting polynomial found was a second order polynomial. The nonlinearity for Sample 3 ranged from -5.2% to 3.8% and from -0.1 IU/mL to 0.6 IU/mL, fulfilling the acceptance criteria.
| Serum
Samples | Test Range
(IU/mL) | Slope
(95% CI) | Y-Intercept
(95% CI) | R² | Average %
Recovery |
|------------------|-----------------------|-----------------------|-------------------------|------|-----------------------|
| 1 | 60.5 – 544.1 | 0.92
(0.88 – 0.96) | 25.9
(11.9 – 39.9) | 0.99 | 104.0% |
| 2 | 10.8 – 108.3 | 1.04
(0.99 - 1.10) | 0.6
(-2.8 - 4.0) | 0.99 | 105.6% |
| 3 | 1.5 - 15.3 | 1.00
(0.95 - 1.05) | -0.6
(-1.1 – -0.1) | 0.99 | 91.2% |
| 4 | 0.2 – 1.5 | 1.07
(0.99 - 1.14) | -0.1
(-0.1 - 0.0) | 0.98 | 96.7% |
| Combined | 0.2 – 544.1 | 0.98
(0.97 – 0.99) | 2.7
(0.5 - 4.9) | 1.00 | 99.3% |
All four samples showed dilution linearity individually and in combination.
For the QUANTA Flash RF IgA, all samples have been found that the best fitting polynomial is a linear one except Sample 1, where the best fitting polynomial found was a third order polynomial. The nonlinearity for Sample 1 ranged from -10.8% to 5.6%, fulfilling the acceptance criteria.
| Serum
Samples | Test Range
(CU) | Slope
(95% CI) | Y-Intercept
(95% CI) | R 2 | Average %
Recovery |
|------------------|--------------------|-----------------------|-------------------------|------|-----------------------|
| 1 | 104.5 - 1045.0 | 1.00
(0.96 - 1.04) | 12.8
(-12.4 - 38.1) | 0.99 | 102.9% |
| 2 | 10.8 - 107.7 | 0.94
(0.90 - 0.98) | 5.1
(2.7 - 7.6) | 0.99 | 106.6% |
| 3 | 1.1 - 11.1 | 0.97
(0.92 - 1.02) | 0.3
(-0.1 - 0.6) | 0.99 | 104.9% |
| Combined | 1.1 - 1045.0 | 1.01
(1.00 - 1.02) | 1.7
(-2.8 - 6.2) | 1.00 | 104.8% |
All three samples showed dilution linearity individually and in combination.
These data demonstrate the linearity of the analytical measuring range (0.3 IU/mL – 490.0 IU/mL) of the QUANTA Flash RF IgM assay and the analytical measuring range (1.3 CU – 900.0 CU) of the QUANTA Flash RF IgA assay.
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Interference
The interference study was performed according to CLSI EPO7-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition. A set of four human serum specimens, one high positive, one low positive, one near the cutoff and one negative sample were tested using the following interfering substances (bilirubin, hemoglobin, triglycerides, cholesterol, human lgG, ascorbic acid, methotrexate and prednisone). All interferents were spiked into every specimen in 10% of total specimen volume, and the resulting samples were assessed in triplicates with the QUANTA Flash RF IgM and the QUANTA Flash RF IgA assays. Recovery of the unit values was calculated compared to control samples spiked with the same volume of diluents (10% of total sample volume). Acceptance criteria for the interference studies were 85% - 115% recovery, or ± 15% of the cut-off (±0.75 IU/mL or ±3 CU) difference, whichever is greater.
No interference was detected with any of the assays with bilirubin up to 1 mg/mL (recovery: from 89.0% to 98.2% and from 92.5 to 93.1 for the IgM and IgA subtypes respectively), hemoglobin up to 2 mg/mL (recovery: from 94.5% to 95.0% and from 90.9 to 94.7 for the IgM and IgA subtypes respectively), triglycerides up to 1000 mg/dL (recovery: from 91.2% to 112.2% and from 97.8 to 105.4 for the IgM and lgA subtypes respectively), cholesterol up to 332.5 mg/dL (recovery: from 89.7% to 91.6% and from 91.0 to 91.0 for the IgM and IgA subtypes respectively), human IgG up to 70 mg/mL (recovery: from 86.8% to 89.3% and from 91.6 to 95.7 for the IgA subtypes respectively), ascorbic acid up to 60.1 mg/L (recovery: from 95.6% to 101.7% and from 98.1 to 102.6 for the IgA subtypes respectively), methotrexate up to 9.1 mg/mL (recovery: from 96.1% to 102.8% and from 92.9 to 105.8 for the IgM and lgA subtypes respectively), and prednisone up to 0.3 mg/L (recovery: from 101.9% to 109.5% and from 93.0 to 94.9 for the IgM and IgA subtypes respectively).
Sample Stability and Handling
For the QUANTA Flash RF IgM, five human serum samples, encompassing negative (n=1), around the cut-off (n=1), and positive samples (n=3), and for the QUANTA Flash RF IgA four human serum samples, encompassing negative (n=1), around the cut-off (n=1), and positive samples (n=2), were tested in duplicates for up to 21 days while stored at 2-8°C, up to 48 hours while stored at room temperature, and after repeated freeze/thaw cycles up to 3 cycles.
Results were compared to those obtained on control samples (time zero / zero cycles).
Acceptance criteria: recovery is between 85-115% for positive samples, and between 80-120% for negative samples (