(263 days)
Not Found
No
The description focuses on a standard chemiluminescent immunoassay and automated instrument processing, with no mention of AI or ML algorithms for data analysis or interpretation.
No.
Explanation: The device is a diagnostic tool designed to measure fecal calprotectin levels, which can aid in the diagnosis and differentiation of Inflammatory Bowel Disease (IBD) from Irritable Bowel Syndrome (IBS). It does not provide any treatment or therapy.
Yes
Explanation: The "Intended Use / Indications for Use" section explicitly states that the device "can aid in the diagnosis of inflammatory bowel disease (IBD) (ulcerative colitis and Crohn's disease), and in the differentiation of IBD from irritable bowel syndrome (IBS)." This directly indicates its role in the diagnostic process.
No
The device is a chemiluminescent immunoassay that runs on a fully automated closed system instrument (BIO-FLASH®) which includes hardware components like liquid handling, a luminometer, and a computer with software. While software is part of the system, the device itself is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it's for the "quantitative determination of fecal calprotectin in extracted human stool samples" to "aid in the diagnosis of inflammatory bowel disease (IBD)... and in the differentiation of IBD from irritable bowel syndrome (IBS)." This clearly indicates it's used to test human specimens (stool) to provide information for diagnostic purposes.
- Device Description: The description details a "chemiluminescent immunoassay" that runs on an automated instrument, utilizing reagents and calibrators to measure a specific analyte (calprotectin) in a biological sample. This is a typical description of an in vitro diagnostic assay.
- Components: The description mentions "QUANTA Flash Calprotectin Reagents," "Calibrators," "Controls," and "Extraction Buffer," all of which are components of an IVD test kit.
- Performance Studies: The document includes detailed performance studies, including analytical and clinical performance, which are required for the validation of an IVD.
- Predicate Device: The mention of a "Predicate Device(s)" with a K number (K160447) indicates a comparison to a previously cleared IVD device, a common practice in the regulatory process for IVDs.
All these factors strongly indicate that the QUANTA Flash Calprotectin is an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
QUANTA Flash Calprotectin is a chemiluminescent immunoassay for the quantitative determination of fecal calprotectin in extracted human stool samples. Elevated levels of fecal calprotectin, in conjunction with clinical findings and other laboratory tests, can aid in the diagnosis of inflammatory bowel disease (IBD) (ulcerative colitis and Crohn's disease), and in the differentiation of IBD from irritable bowel syndrome (IBS).
QUANTA Flash Calprotectin Calibrators are intended for use with the QUANTA Flash Calprotectin Reagents for the determination of fecal calprotectin levels in extracted stool samples. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash Calprotectin Controls are intended for use with the QUANTA Flash Calprotectin Reagents for quality control in the determination of fecal calprotectin levels in extracted stool samples.
QUANTA Flash Calprotectin Extraction Buffer is intended for use with the QUANTA Flash Calprotectin Reagents as sample extraction solution.
Product codes (comma separated list FDA assigned to the subject device)
NXO, JIT, JJX
Device Description
The principle of the assay is chemiluminescent microparticle immunoassay, a variation of solid phase immunoassay. The QUANTA Flash® Calprotectin 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® Calprotectin assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH® instrument.
Calprotectin-specific capture antibodies are coated on to paramagnetic beads, which are stored in the reagent cartridge under conditions that preserve the antibody in its reactive state. Prior to use in the BIO-FLASH® system, the reagent pack containing all the necessary assay reagents is mixed thoroughly by being inverted several times. The sealed reagent tubes are pierced with the reagent cartridge lid, and the reagent cartridge is loaded onto the instrument. Reagents are calibrated when the lot is first used. A patient extracted stool sample is prediluted by the BIO-FLASH® with sample buffer in a disposable plastic cuvette. Small amounts of the diluted patient extracted stool, the beads, and the 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 monoclonal 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 (urea-hydrogen peroxide in sodium chloride solution) are added to the cuvette, and the flash of light produced from this reaction is measured as Relative Light Units (RLU) by the BIO-FLASH® optical system. The measured RLU is proportional to the amount of bound isoluminol conjugate, which is in turn proportional to the amount of calprotectin antigen captured by the antibodies (anti-calprotectin polyclonal antibodies in this case) on the beads. For quantitation, the QUANTA Flash® Calprotectin will utilize a predefined lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. The Master Curve is generated by Inova Diagnostics for each reagent pack lot with in-house Standards with assigned unit values (ng/mL). The RLU and assigned ng/mL values of the Standards are used to create a 4 parameter logistic curve. These four parameters are embedded in the reagent pack barcode. When the lot is used the first time, the Calibrators are run, and based on the results obtained on the Calibrators, an instrument specific Working Curve is created; The Working Curve is used to calculate units (ng/mL) based on RLU values obtained on each sample. The obtained ng/mL values will be converted to mg/kg by a calculation that takes into account the dilution of the samples. This unit conversion is calculated automatically by the software.
The QUANTA Flash Calprotectin Reagents kit contains the following materials:
One (1) QUANTA Flash Calprotectin Reagent Cartridge
One (1) bottle of QUANTA Flash Special Wash
The QUANTA Flash Calprotectin reagent cartridge contains the following reagents for 100 determinations:
- a. Anti- calprotectin antibodies coated paramagnetic beads.
- b. Assay buffer - colored pink, containing Tris-buffered saline, Tween 20, protein stabilizers and preservatives.
- c. Tracer anti-calprotectin Isoluminol labeled anti-calprotectin monoclonal antibodies in buffer, containing protein stabilizers and preservative.
The QUANTA Flash Calprotectin Calibrators kit contains two vials each of Calibrator 2, and Calibrator 3:
QUANTA Flash Calprotectin Calibrators:
- QUANTA Flash Calprotectin Calibrator 1: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain recombinant calprotectin antigen in stabilizers and preservatives.
- QUANTA Flash Calprotectin Calibrator 2: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain recombinant calprotectin antigen in stabilizers and preservatives.
- QUANTA Flash Calprotectin Calibrator 3: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain recombinant calprotectin antigen in stabilizers and preservatives.
The QUANTA Flash Calprotectin Controls kit contains two vials of Low Control and two vials of High Control:
QUANTA Flash Calprotectin Controls:
- QUANTA Flash Calprotectin Low Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain recombinant calprotectin antigen in stabilizers and preservatives.
- QUANTA Flash Calprotectin High Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain recombinant calprotectin antigen in stabilizers, and preservatives.
The QUANTA Flash Calprotectin Extraction Buffer kit contains two bottles of Extraction Buffer (2.5 X):
QUANTA Flash Calprotectin Extraction Buffer:
- QUANTA Flash Calprotectin Extraction Buffer (2.5 X): Two (2) labeled bottles containing 125 mL, concentrated reagent.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Human stool samples (fecal)
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 Calprotectin. A total of 165 characterized samples were included in the Validation Set for the QUANTA Flash Calprotectin. Samples came from studies performed at two different sites. Samples with ID number from 1 to 107 belong to site A, while samples with ID number from 108 to 175 belong to site B.
For site A: Inclusion criteria: samples suspected to suffer from IBD and that a calprotectin test was requested. Samples were consecutive and recruited over a period of 6 months. To allow accurate diagnostic, all patients underwent ileocolonoscopy. Exclusion criteria: unclear diagnosis (e.g. indeterminate colitis), inability to collect enough fecal samples and age younger of 14 years. Also patients that previously had been diagnosed IBD or had not received ileocolonoscopy were excluded. IBD diagnostic criteria: diagnostic work-up included physical examination and case history, endoscopic and histologic analysis, radiologic work-up and laboratory tests including ileocolonoscopy. Senior gastroenterologists performed all endoscopies and findings were documented in a computer-based database. The final diagnosis of IBD (i.e. CD and UC) was independently made by a pathologist or gastroenterologist who was blinded for calprotectin results.
For site B: Sixty-eight consecutive samples requested for a calprotectin determination test by a gastroenterologist who had not had a calprotectin level measured before. Exclusion criteria: samples with excessive mucous, unclear diagnosis, patients who had not received colonoscopy. IBD diagnostic criteria: Patients were diagnosed after exclusion of organic pathology on the basis of routine blood tests, thyroid function tests, serological screening for coeliac disease, stool examination for bacteria and parasites, ultrasound examination, and eventually colonoscopy using the ROME III criteria. The diagnosis of IBD was made upon clinical, endoscopic and histological findings as described in J Crohn Colitis 2012:6: 965-990 (ulcerative colitis) and J Crohn Colitis 2010:4:7-27 (Crohn's disease).
All samples were run on the QUANTA Flash Calprotectin.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision Study:
The precision of the QUANTA Flash Calprotectin assay was evaluated on 8 samples containing various concentrations of calprotectin antigen in accordance with CLSI EPO5-A3, Evaluation of Precision Performance of Quantitative Measurement Procedures - Approved Guideline. Samples were run in duplicates, twice a day, for 20 days.
Sample ID, N, Mean (mg/kg), Repeatability SD (mg/kg), Repeatability CV (%), Between-Run SD (mg/kg), Between-Run CV (%), Between-Day SD (mg/kg), Between-Day CV (%), Within-Laboratory Precision SD (mg/kg), Within-Laboratory Precision CV (%)
1, 80, 51.1, 2.1, 4.0%, 1.9, 3.6%, 0.5, 1.0%, 2.8, 5.5%
2, 80, 72.2, 2.1, 3.0%, 1.5, 2.0%, 0.7, 0.9%, 2.7, 3.7%
3, 80, 108.1, 2.5, 2.3%, 2.5, 2.3%, 0.0, 0.0%, 3.6, 3.3%
4, 80, 104.2, 2.4, 2.3%, 1.4, 1.3%, 1.6, 1.5%, 3.2, 3.1%
5, 80, 196.0, 5.8, 2.9%, 4.5, 2.3%, 1.1, 0.5%, 7.4, 3.8%
6, 80, 639.5, 18.3, 2.9%, 11.1, 1.7%, 4.3, 0.7%, 21.9, 3.4%
7, 80, 1086.9, 34.2, 3.1%, 27.0, 2.5%, 0.0, 0.0%, 43.6, 4.0%
8, 80, 1828.0, 58.2, 3.2%, 33.8, 1.8%, 44.9, 2.5%, 80.8, 4.4%
9, 80, 43.1, 1.6, 3.8%, 1.7, 4.0%, 1.2, 2.9%, 2.7, 6.2%
10, 80, 3036.3, 90.2, 3.0%, 120.3, 4.0%, 46.2, 1.5%, 157.3, 5.2%
Acceptance criteria: Total %CV:
§ 866.5180 Fecal calprotectin immunological test system.
(a)
Identification. A fecal calprotectin immunological test system is anin vitro diagnostic device that consists of reagents used to quantitatively measure, by immunochemical techniques, fecal calprotectin in human stool specimens. The device is intended forin vitro diagnostic use as an aid in the diagnosis of inflammatory bowel diseases (IBD), specifically Crohn's disease and ulcerative colitis, and as an aid in differentiation of IBD from irritable bowel syndrome.(b)
Classification. Class II (special controls). The special control for these devices is FDA's guidance document entitled “Class II Special Controls Guidance Document: Fecal Calprotectin Immunological Test Systems.” For the availability of this guidance document, see § 866.1(e).
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December 22, 2018 Inova Diagnostics, Inc. Roger Albesa Supervisor. Research and Development 9900 Old Grove Road San Diego, California 92131-1638
Re: K170993
Trade/Device Name: QUANTA Flash Calprotectin Reagents, QUANTA Flash Calprotectin Calibrators, OUANTA Flash Calprotectin Controls, OUANTA Flash Calprotectin Extraction Buffer Regulation Number: 21 CFR 866.5180 Regulation Name: Fecal calprotectin immunological test system Regulatory Class: Class II Product Code: NXO, JIT Dated: November 21, 2017 Received: November 22, 2017
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. 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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR
1
803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); 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 the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
For comprehensive regulatory information about mediation-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.
Kelly Olir
For,
Lea Carrington 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)
Device Name
QUANTA Flash Calprotectin, QUANTA Flash Calibrators, QUANTA Flash Calprotectin Controls, QUANTA Flash Calprotectin Extraction Buffer
Indications for Use (Describe)
QUANTA Flash Calprotectin is a chemiluminescent immunoassay for the quantitative determination of fecal calprotectin in extracted human stool samples. Elevated levels of fecal calprotectin, in conjunction with clinical findings and other laboratory tests, can aid in the diagnosis of inflammatory bowel disease (IBD) (ulcerative colitis and Crohn's disease), and in the differentiation of IBD from irritable bowel syndrome (IBS).
QUANTA Flash Calprotectin Calibrators are intended for use with the QUANTA Flash Calprotectin Reagents for the determination of fecal calprotectin levels in extracted stool samples. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash Calprotectin Controls are intended for use with the QUANTA Flash Calprotectin Reagents for quality control in the determination of fecal calprotectin levels in extracted stool samples.
| 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® Calprotectin Reagents | |
|---|---|
| QUANTA Flash® Calprotectin Calibrators | |
| QUANTA Flash® Calprotectin Controls | |
| QUANTA Flash® Calprotectin Extraction Buffer | Page 1 of 23 |
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 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 |
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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® Calprotectin Reagents
QUANTA Flash® Calprotectin Calibrators
QUANTA Flash® Calprotectin Controls
QUANTA Flash® Calprotectin Extraction Buffer | |
| Scientific contact: | Roger Albesa, Supervisor, 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: 858-586-9900/1381
Fax: 858-863-0025
Email: relliot@inovadx.com | |
| Device name (assay kit): | Proprietary name: | QUANTA Flash® Calprotectin Reagents |
| | Common name: | Fecal Calprotectin Chemiluminescent
Immunoassay |
| | Classification name: | Calprotectin, Fecal |
| Regulation Description | Fecal Calprotectin Immunological Test System | |
| Regulation Medical Specialty | Immunology | |
| Review Panel | Immunology | |
| Product Code | NXO | |
| Regulation Number | 866.5180 | |
| Device Class | 2 | |
| Device name (Calibrators): | Proprietary name: QUANTA Flash® Calprotectin Calibrators
Common name: Calprotectin 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® Calprotectin Controls
Common name: Calprotectin 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) | |
| Device name (Extract. Buffer): | Proprietary name: QUANTA Flash® Calprotectin Extraction Buffer
Common name: Calprotectin Extraction Solution
Classification name: Calprotectin, Fecal | |
| Regulation Description | Fecal Calprotectin Immunological Test System | |
| Regulation Medical Specialty | Immunology | |
| Product Code | NXO | |
| Regulation Number | 866.5180 | |
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510(k) Summary QUANTA Flash® Calprotectin
6
Predicate device
Calprest NG (QUANTA Lite® Calprotectin Extended Range ELISA), 510(k) number: K160447. Date declared: November 10, 2016.
Device description
The principle of the assay is chemiluminescent microparticle immunoassay, a variation of solid phase immunoassay. The QUANTA Flash® Calprotectin 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® Calprotectin assay utilizes a reagent cartridge format, which is compatible with the BIO-FLASH® instrument.
Calprotectin-specific capture antibodies are coated on to paramagnetic beads, which are stored in the reagent cartridge under conditions that preserve the antibody in its reactive state. Prior to use in the BIO-FLASH® system, the reagent pack containing all the necessary assay reagents is mixed thoroughly by being inverted several times. The sealed reagent tubes are pierced with the reagent cartridge lid, and the reagent cartridge is loaded onto the instrument. Reagents are calibrated when the lot is first used. A patient extracted stool sample is prediluted by the BIO-FLASH® with sample buffer in a disposable plastic cuvette. Small amounts of the diluted patient extracted stool, the beads, and the 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 monoclonal 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 (urea-hydrogen peroxide in sodium chloride solution) are added to the cuvette, and the flash of light produced from this reaction is measured as Relative Light Units (RLU) by the BIO-FLASH® optical system. The measured RLU is proportional to the amount of bound isoluminol conjugate, which is in turn proportional to the amount of calprotectin antigen captured by the antibodies (anti-calprotectin polyclonal antibodies in this case) on the beads. For quantitation, the QUANTA Flash® Calprotectin will utilize a predefined lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. The Master Curve is generated by Inova Diagnostics for each reagent pack lot with in-house Standards with assigned unit values (ng/mL). The RLU and assigned ng/mL values of the Standards are used to create a 4 parameter logistic curve. These four parameters are embedded in the reagent pack barcode. When the lot is used the first time, the Calibrators are run, and based on the results obtained on the Calibrators, an instrument specific Working Curve is created; The Working Curve is used to calculate units (ng/mL) based on RLU values obtained on each sample. The obtained ng/mL values will be converted to mg/kg by a calculation that takes into account the dilution of the samples. This unit conversion is calculated automatically by the software.
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The QUANTA Flash Calprotectin Reagents kit contains the following materials:
One (1) QUANTA Flash Calprotectin Reagent Cartridge
One (1) bottle of QUANTA Flash Special Wash
The QUANTA Flash Calprotectin reagent cartridge contains the following reagents for 100 determinations:
- a. Anti- calprotectin antibodies coated paramagnetic beads.
- b. Assay buffer - colored pink, containing Tris-buffered saline, Tween 20, protein stabilizers and preservatives.
- Tracer anti-calprotectin Isoluminol labeled anti-calprotectin monoclonal antibodies in C. buffer, containing protein stabilizers and preservative.
The QUANTA Flash Calprotectin Calibrators kit contains two vials each of Calibrator 2, and Calibrator 3:
QUANTA Flash Calprotectin Calibrators:
- QUANTA Flash Calprotectin Calibrator 1: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain recombinant calprotectin antigen in stabilizers and preservatives.
- -QUANTA Flash Calprotectin Calibrator 2: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain recombinant calprotectin antigen in stabilizers and preservatives.
- -QUANTA Flash Calprotectin Calibrator 3: Two (2) barcode labeled tubes containing 0.3 mL prediluted, ready to use reagent. Calibrators contain recombinant calprotectin antigen in stabilizers and preservatives.
The QUANTA Flash Calprotectin Controls kit contains two vials of Low Control and two vials of High Control:
QUANTA Flash Calprotectin Controls:
- । QUANTA Flash Calprotectin Low Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain recombinant calprotectin antigen in stabilizers and preservatives.
- । QUANTA Flash Calprotectin High Control: Two (2) barcode labeled tubes containing 0.5 mL, ready to use reagent. Controls contain recombinant calprotectin antigen in stabilizers, and preservatives.
The QUANTA Flash Calprotectin Extraction Buffer kit contains two bottles of Extraction Buffer (2.5 X): QUANTA Flash Calprotectin Extraction Buffer:
- QUANTA Flash Calprotectin Extraction Buffer (2.5 X): Two (2) labeled bottles containing 125 । mL, concentrated reagent.
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Intended use(s)
QUANTA Flash Calprotectin is a chemiluminescent immunoassay for the quantitative determination of fecal calprotectin in extracted human stool samples. Elevated levels of fecal calprotectin, in conjunction with clinical findings and other laboratory tests, can aid in the diagnosis of inflammatory bowel disease (IBD) (ulcerative colitis and Crohn's disease), and in the differentiation of IBD from irritable bowel syndrome (IBS).
QUANTA Flash Calprotectin Calibrators are intended for use with the QUANTA Flash Calprotectin Reagents for the determination of fecal calprotectin levels in extracted stool samples. Each calibrator establishes a point of reference for the working curve that is used to calculate unit values.
QUANTA Flash Calprotectin Controls are intended for use with the QUANTA Flash Calprotectin Reagents for quality control in the determination of fecal calprotectin levels in extracted stool samples.
QUANTA Flash Calprotectin Extraction Buffer is intended for use with the QUANTA Flash Calprotectin Reagents as sample extraction solution.
Indications for use
Same as Intended use.
Substantial equivalence
The QUANTA Flash Calprotectin Reagents, the QUANTA Flash Calprotectin Calibrators, the QUANTA Flash Calprotectin Controls and the QUANTA Flash Calprotectin Extraction Buffer have the same intended use and assay principle as the predicate device.
Comparison to predicate device
| Similarities | ||
|---|---|---|
| Item | QUANTA Flash Calprotectin Reagents | QUANTA Lite Calprotectin Extended Range ELISA |
| Intended use | QUANTA Flash Calprotectin is a chemiluminescent immunoassay for the quantitative determination of fecal calprotectin in extracted human stool samples. Elevated levels of fecal calprotectin, in conjunction with clinical findings and other laboratory tests, can aid in the diagnosis of inflammatory bowel disease (IBD) (ulcerative colitis and Crohn's disease), and in the | QUANTA Lite Calprotectin Extended Range is a quantitative ELISA for detecting concentration of fecal calprotectin, which can be used as an in vitro diagnostic to aid in the diagnosis of Inflammatory Bowel Diseases (IBD), specifically Crohn's disease and ulcerative colitis, and to differentiate IBD from Irritable Bowel Syndrome (IBS) |
QUANTA Flash Calprotectin Reagents
9
| Similarities | ||
|---|---|---|
| ltem | QUANTA Flash Calprotectin Reagents | QUANTA Lite Calprotectin Extended |
| Range ELISA | ||
| differentiation of IBD from irritable bowel | in conjunction with other clinical and | |
| syndrome (IBS). | laboratory findings. | |
| Assay methodology | Solid phase (heterogeneous) | Solid phase (heterogeneous) |
| immunoassay | immunoassay | |
| Antigen | Rabbit polyclonal anti-calprotectin | Rabbit polyclonal anti-calprotectin |
| antibody | antibody | |
| Shelf life | One year | One year |
| Sample type | Extracted Human Stool | Extracted Human Stool |
| Units | mg/kg (milligram of calprotectin per | mg/kg (milligram of calprotectin per |
| kilogram of stool) | kilogram of stool) |
| Differences | ||
|---|---|---|
| Item | QUANTA Flash Calprotectin Reagents | QUANTA Lite Calprotectin Extended Range ELISA |
| Detection/ | ||
| Operating principle | Chemiluminescent immunoassay | Enzyme-linked immunosorbent assay |
| Solid phase | Paramagnetic microparticles (beads) | 96-well polystyrene plate |
| Conjugate | Isoluminol conjugated monoclonal anti- | |
| calprotectin antibody | HRP conjugated monoclonal anti- | |
| calprotectin antibody | ||
| Analytical | ||
| Measuring Range | 16.1 – 3,500.0 mg/kg | 27.1 – 3,000.0 mg/kg |
| Calibration | Lot specific Master Curve + three | |
| calibrators (sold separately) | Six lot specific calibrators |
QUANTA Flash Calprotectin Calibrators
| Item | QUANTA Flash Calprotectin Calibrators | QUANTA Lite Calprotectin Extended
Range ELISA |
|--------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------|
| Intended use | QUANTA Flash Calprotectin Calibrators
are intended for use with the QUANTA
Flash Calprotectin Reagents for the
determination of fecal calprotectin
levels in extracted stool samples. 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 | Recombinant calprotectin antigen (rAg) | Recombinant calprotectin antigen (rAg) |
| Method | QUANTA Flash Calprotectin
chemiluminescent immunoassay | QUANTA Lite Calprotectin Extended
Range ELISA |
| Matrix | Calprotectin rAg, stabilizer, and
preservative | Calprotectin rAg, stabilizer, and
preservative |
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| Item | QUANTA Flash Calprotectin Calibrators | QUANTA Lite Calprotectin Extended |
|---|---|---|
| Range ELISA | ||
| Units | ng/mL | ng/mL |
| 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 Calprotectin Controls
| Item | QUANTA Flash Calprotectin Controls | QUANTA Lite Calprotectin Extended
Range ELISA |
|-------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------|
| Intended use | QUANTA Flash Calprotectin Controls
are intended for use with the QUANTA
Flash Calprotectin Reagents for quality
control in the determination of fecal
calprotectin levels in extracted stool
samples. | No separate intended use; controls are
part of the kit. |
| Analyte | Recombinant calprotectin antigen (rAg) | Recombinant calprotectin antigen (rAg) |
| Method | QUANTA Flash Calprotectin
chemiluminescent immunoassay | QUANTA Lite Calprotectin Extended
Range ELISA |
| Matrix | Calprotectin rAg, stabilizer, and
preservative | Calprotectin rAg, stabilizer, and
preservative |
| Units | ng/mL | ng/mL |
| Physico-chemical
characteristics | Liquid, ready to use | Liquid, prediluted, ready to use |
| Levels | 2 (low and high) | 2 (1 and 2) |
| Storage | 2-8 °C | 2-8 °C |
| Shelf life | One year | One year |
QUANTA Flash Calprotectin Extraction Buffer
| ltem | QUANTA Flash Calprotectin Extraction
Buffer | QUANTA Lite Calprotectin Extended
Range ELISA |
|------------------|------------------------------------------------|--------------------------------------------------|
| | | |
| Intended use | QUANTA Flash Calprotectin Extraction | |
| | Buffer is intended for use with the | No separate intended use; controls are |
| | QUANTA Flash Calprotectin Reagents as | part of the kit. |
| | sample extraction solution. | |
| Method | QUANTA Flash Calprotectin | QUANTA Lite Calprotectin Extended |
| | chemiluminescent immunoassay | Range ELISA |
| Concentration | 2.5X (1X working concentration) | 2.5X (1X working concentration) |
| Physico-chemical | | |
| characteristics | Liquid, ready to use | Liquid, ready to use |
| Storage | 2-8 °C | 2-8 °C |
| Shelf life | One year | One year |
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Analytical performance characteristics
Quantitation and units of measure
For quantitation, the QUANTA Flash Calprotectin assay utilizes a lot specific Master Curve that is uploaded onto the instrument through the reagent cartridge barcode. The Master Curve for QUANTA Flash Calprotectin consists of 7 Standards. These Master Curve Standards are used to create the lot specific Master Curve during the manufacturing procedure.
List of Calprotectin Standards:
| Material | Assigned Value |
|---|---|
| Calprotectin Master Curve Standard 1 | 0.0 ng/mL |
| Calprotectin Master Curve Standard 2 | 10.9 ng/mL |
| Calprotectin Master Curve Standard 3 | 43.5 ng/mL |
| Calprotectin Master Curve Standard 4 | 173.9 ng/mL |
| Calprotectin Master Curve Standard 5 | 695.6 ng/mL |
| Calprotectin Master Curve Standard 6 | 1391.3 ng/mL |
| Calprotectin Master Curve Standard 7 | 3478.3 ng/mL |
Value assignment and traceability of Calibrators and Controls
The QUANTA Flash Calprotectin Calibrators and Controls are manufactured by diluting recombinant calprotectin antigen in a buffer with stabilizers and preservatives. The recombinant calprotectin antigen is obtained from commercial sources.
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 5 to obtain a minimum of 10 data points 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 Calprotectin assay.
| Material | Manufacturing
Target Value | Manufacturing
Target Range |
|---------------------------|-------------------------------|-------------------------------|
| Calprotectin Calibrator 1 | 40 ng/mL | 32 – 48 ng/mL |
| Calprotectin Calibrator 2 | 800 ng/mL | 640 – 960 ng/mL |
| Calprotectin Calibrator 3 | 2400 ng/mL | 2160 – 2640 ng/mL |
| Calprotectin Low Control | 40 ng/mL | 32 – 48 ng/mL |
| Calprotectin High Control | 200 ng/mL | 180 – 220 ng/mL |
Calprotectin Calibrators and Controls with target manufacturing values:
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Precision
The precision of the QUANTA Flash Calprotectin assay was evaluated on 8 samples containing various concentrations of calprotectin antigen in accordance with CLSI EPO5-A3, Evaluation of Precision Performance 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: 3,500.0 mg/kg after further diluting it by 10 fold,
15
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 3,500.0 mg/kg, the highest value that can be reported is 35,000.0 mg/kg.
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 when tested as neat samples). RLU values showed increase with increasing antibody concentrations above the AMR, thereby confirming that high positive specimens above the AMR do not show hook effect up to 21,753.6 mg/kg (theoretical value calculated using the highest value in the AMR and its dilution factor) in the QUANTA Flash Calprotectin assay.
Linearity
The linearity of the AMR of the QUANTA Flash Calprotectin 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 in two ways: using extracted stool samples and using recombinant antigen samples. Three extracted stool samples calprotectin antigen concentrations were combined with another extracted stool sample containing low levels of calprotectin antigen in 10% increments (from 0% to 90% of low sample) to obtain values that cover the entire AMR. Additionally, three samples made by diluting recombinant antigen in buffer containing stabilizers and preservatives (same buffer used to make controls and calibrators) containing various calprotectin recombinant antigen concentrations were diluted in buffer in 10% increments (from 0% to 90% of buffer) to obtain values that cover the entire AMR. The dilutions were assayed in duplicates. Results were analyzed according to the guideline performing regression analysis and identifying the best fitting polynomial.
Acceptance criteria:
- Best fitting polynomial is a linear one, otherwise, the difference between the best-fitting nonlinear and linear polynomial is less than 15% (allowable nonlinearity).
All samples (3 stool samples and 3 recombinant antigen samples) have been found that the best fitting polynomial is a linear one except rAg Sample 2, where the best fitting polynomial found was a second order polynomial. The nonlinearity for rAg sample 2 ranged from -13.0% to 2.1%, fulfilling the acceptance criteria.
16
| Stool
Sample | Test Range
(mg/kg) | Slope
(95% CI) | Y-Intercept
(95% CI) | R2 | Average %
Recovery |
|-----------------|-----------------------|------------------------|--------------------------|------|-----------------------|
| 1 | 4102.8 to 410.3 | 1.02
(0.97 to 1.06) | 16.1
(-90.1 to 122.3) | 1.00 | 102.6% |
| 2 | 890.3 to 89.0 | 0.95
(0.90 to 1.00) | 11.9
(-14.2 to 38.0) | 1.00 | 100.0% |
| 3 | 155.6 to 15.6 | 1.12
(0.98 to 126) | -5.5
(-19.0 to 8.0) | 0.98 | 100.8% |
| Combined | 4102.8 to 15.6 | 1.02
(1.01 to 1.03) | -5.8
(-25.6 to 14.1) | 1.00 | 101.1% |
Moreover, a regression analysis has been performed in all individual samples and in combination of them, obtaining the results summarized in the tables below:
| All three extracted stool samples showed dilution linearity individually and in combination. | |
|---|---|
| ---------------------------------------------------------------------------------------------- | -- |
| rAg
Sample | Test Range
(ng/mL) | Slope
(95% CI) | Y-Intercept
(95% CI) | R2 | Average %
Recovery |
|---------------|-----------------------|------------------------|---------------------------|------|-----------------------|
| 1 | 3376.4 to 337.6 | 1.03
(0.97 to 1.09) | -85.9
(-209.5 to 37.6) | 1.00 | 96.8% |
| 2 | 618.9 to 61.9 | 0.98
(0.95 to 1.02) | 15.9
(3.0 to 28.7) | 1.00 | 104.7% |
| 3 | 115.1 to 11.5 | 0.99
(0.93 to 1.05) | 4.0
(-0.6 to 8.5) | 0.99 | 108.7% |
| Combined | 3376.4 to 11.5 | 1.00
(0.98 to 1.02) | -2.9
(-26.0 to 20.2) | 1.00 | 103.4% |
These data demonstrate the linearity of the analytical measuring range (14.0 ng/mL – 3,043.5 ng/mL / 16.1 mg/kg - 3,500.0 mg/kg) of the QUANTA Flash Calprotectin assay.
Recovery
The recovery of the QUANTA Flash Calprotectin assay has been evaluated using seven extracted stool samples containing various concentrations of calprotectin antigen covering the AMR of the assay. Samples were spiked with calibrator material and then tested to calculate recovery results. For the samples with calprotectin concentrations lower than 200 mg/kg, Calibrator 2 was used as spiking material, while for samples with concentrations higher than 200 mg/kg, Calibrator 3 was used. Each sample was mixed with their correspondent calibrator material in a proportion 9:1. Each sample, calibrator and spiked sample was tested in duplicate and recovery values were calculated.
Acceptance criteria: Percent Recovery must be between 88% and 112%
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| Theoretical Value | Observed Value | ||||
|---|---|---|---|---|---|
| Sample | Baseline | Calibrator Value | 90% sample + 10% Calibrator | 90% sample + 10% Calibrator | Recovery |
| (mg/kg) | (mg/kg) | (mg/kg) | (mg/kg) | % | |
| Sample 1 | 49.6 | 740.6 | 118.7 | 128.3 | 108.1% |
| Sample 2 | 83.1 | 740.6 | 148.8 | 141.9 | 95.4% |
| Sample 3 | 155.3 | 740.6 | 213.8 | 212.7 | 99.5% |
| Sample 4 | 126.9 | 740.6 | 188.2 | 202.3 | 107.5% |
| Sample 5 | 252.3 | 2624.4 | 489.5 | 518.4 | 105.9% |
| Sample 6 | 833.2 | 2624.4 | 1012.3 | 1112.0 | 109.8% |
| Sample 7 | 2501.6 | 2624.4 | 2513.9 | 2380.0 | 94.7% |
All recovery results fulfilled the acceptance criteria and ranged from 94.7% to 109.8% and are summarized in the table below:
Interference
The interference study was performed according to CLSI EPO7-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition. Six human stool specimens, one high positive (1365.1 mg/kg), one moderately positive (664.7 mg/kg), one low positive (215.1 mg/kg), one near the cutoff (123.0 mg/kg), one in the indeterminate rage (62.2 mg/kg) and one negative (22.1 mg/kg), samples were tested. Interfering substances (Hemoglobin, mesalamine, prednisone, vancomycin, gamma-tocopherol, tacrolimus, beta-carotene, ciprofloxacin, cholecalciferol, lansoprazole, and ascorbic acid, along with bacterial cultures: Yersinia ruckeri, Klebsiella pneumoniae, Shigella sonnei, Salmonella enterica and Escherichia coli) were spiked into every specimen in 10% of total specimen volume, and the resulting samples were assessed in triplicates with the QUANTA Flash Calprotectin assay. 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 low indeterminate range (±7.5 mg/kg) difference, whichever is greater.
No interference was detected in the results of the QUANTA Flash Calprotectin at the concentrations tested (hemoglobin 5.56mg/50mg stool, mesalamine 1.33mg/50mg stool, prednisone 0.01mg/50mg stool, vancomycin 0.67mg/50mg stool, gamma-tocopherol 0.0010mg/50mg stool, tacrolimus 0.07mg/50mg stool, beta-carotene 0.0048mg/50mg stool, ciproflaxin 0.50mg/50mg stool, cholecalciferol 0.275ng/50mg stool, lansoprazole 0.02mg/50 mg stool, ascorbic acid 0.05mg/50mg stool respectively for drugs and nutrients and 1.5x10' cfu/mL for each individual bacterial cultures).
Sample Stability and Handling
Eight extracted human stool samples, encompassing negative (n=1), in the indeterminate range (n=2), around the cut-off (n=1), and positive samples (n=4) were tested in duplicates for up to 24 days while stored at 2-8°C, up to 73 hours while stored at room temperature, and after repeated freeze/thaw cycles up to 5 cycles.
18
510(k) Summary QUANTA Flash® Calprotectin
Additionally, three extracted stool samples in the indeterminate range (n=1), around the cut-off (n=1) and high positive (n=1) were tested in triplicates for up to 91 days while frozen at -20±4 ℃.
Results were compared to those obtained on control samples (time zero / zero cycles).
Acceptance criteria: 80-120% average recovery.
All samples fulfilled the acceptance criteria at each time point for each condition. Based on these result, we recommend that extracted samples are stored up to 72 hours at room temperature, up to 21 days at 2-8°C, up to 3 months frozen at -20±5 °C and can be subjected to up to 4 freeze/thaw cycles.
Reagent Stability
Shelf life
To establish the initial claim for shelf life, accelerated stability studies were performed for 3 weeks at 37 °C, where one week is equal to six months at 5 ± 3°C.
Accelerated stability testing was performed on each of the following sealed components of the QUANTA Flash Calprotectin to establish initial stability claim:
| • Calprotectin beads | (3 Lots) |
|---|---|
| • Calprotectin tracer | (3 Lots) |
| • Calibrators 1, 2 and 3 | (3 Lots) |
| • Low and High controls | (3 Lots) |
| • Extraction Buffer | (3 Lots) |
| • Special Wash | (3 Lots) |
Each week a new sealed component was placed in the incubator, and all components were tested at the end of the experiment together with the one that was stored at 5 ± 3°C. The recovery of the measured values was calculated for each time point (compared to those obtained with 5 ± 3°C stored reagent). All calculations were performed by comparing results of sealed components stored at 5 ± 3℃ (control) to those stored at 37 ± 3℃ (test) for 1, 2 and 3 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:
With regression analysis, the lower and upper 95% Cl interval of the regression line is between 80% and 120% recovery at day 14.
All components tested fulfilled the acceptance criteria above, so one year expiration dating was assigned to each component
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In-use (onboard) stability
Calibrators
Onboard stability claim: 4 calibrations, or 8 hours onboard.
During assessing onboard stability, Calibrators were placed uncapped, onboard the instrument, and calibration was performed altogether five times over 9 hours. Controls and a panel of characterized patient specimens were run on each calibration curve.
Calibrators are considered stable if all four calibrations performed in the 8 hour period are successful, mean Calibrator RLU recovery values for the first 4 calibrations are between 90% and 110% compared to the first use, and Control/patient panel ng/mL recovery values are between 85% and 115% of those obtained on the first calibration curve.
The first four calibrations performed in the 8 hour period were considered valid by the software. The calibrators yielded average RLU recovery values ranging from 100.0% to 107.1%. The Control/patient panel ng/mL recovery ranged from 89.1% 105.2%. 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 once a day during 20 days for a total of 20 runs. The first run was used to establish baseline value, by running each vial in duplicate, and then additional 19 runs were performed, by running each vial in singleton. During runs, the Controls were left uncapped, onboard the instrument for 15±1 minutes per run. When not in use, the controls were capped, and stored at 5±3°C.
Percent recovery of each value was calculated compared to the baseline value. Controls are considered stable when all values run within their established range, and the linear regression line obtained by plotting percent recovery values against the number of runs stays between 85% and 15.
All controls ran within their respective acceptable ranges for all runs. Moreover, the regression line remained between 85% and 115% at run 15 for both Controls. These results support the claim that controls can be used for up to 15 times, at 10 minutes per use.
Reagent Cartridge
To establish the in-use stability of the QUANTA Flash Calprotectin reagent cartridge, one lot of reagent cartridge was tested with 4 extracted specimens (with different reactivity levels) along with the 2 controls made of recombinant antigen for a total of 6 samples. The specimens were tested periodically for of 97 days. Percent recoveries were calculated compared to the day zero average values, and linear regression analysis was performed by plotting percent 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 proceeding the day when the 95% confidence interval of the regression line reaches 85% or 115% recovery, or
20
- At the actual measurement day preceding the day when ≥2% of the recovery data, (3 data points) is