AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

The DiaSorin LIAISON® Calprotectin assay is an in vitro diagnostic chemiluminescent immunoassay (CLIA) intended for the quantitative measurement, in human stool, of fecal calprotectin, a neutrophilic protein that is a marker of mucosal inflammation. The LIAISON® Calprotectin assay can be used 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 (IBS). Test results are to be used in conjunction with information obtained from the patients' clinical evaluation and other diagnostic procedures.

The test has to be performed on the LIAISON® XL Analyzer.

The DiaSorin LIAISON® O.S.E.T. Device (Quantitative Stool Extraction and Test) is intended for use in the preparation of human stool specimens for testing in the LIAISON® Calprotectin assay.

Device Description

The LIAISON® Calprotectin assay is a sandwich assay that uses 2 monoclonal antibodies for capture and detection of calprotectin. The LIAISON® Calprotectin assay must be run on the LIAISON® XL Analyzer, a fully automated system with continuous loading.

Calprotectin is first extracted from human stool samples with LIAISON® Q.S.E.T. Buffer using either the weigh method or the LIAISON® Q.S.E.T. Device. The assay incubates extracted sample, calibrator, control, or calibration verifiers with assay buffer and paramagnetic particles coated with a monoclonal antibody that specifically recognizes the calprotectin heterocomplex. Following incubation, a wash cycle is performed to remove any unbound material. An isoluminol conjugated monoclonal antibody that recognizes calprotectin is then added to the reaction and incubated. The unbound conjugate is removed with a second wash step. Starter reagents are then added and a flash chemiluminescent reaction is initiated. The light signal is measured by a photomultiplier as relative light units (RLU) and is proportional to the concentration of calprotectin present in the calibrators, controls or samples.

All assay steps and incubations are performed by the LIAISON® XL Analyzer. The analyzer software automatically calculates the concentration of calprotectin in the sample. This concentration is expressed in ug/q.

AI/ML Overview

Here's an analysis of the acceptance criteria and supporting studies for the DiaSorin LIAISON® Calprotectin assay, LIAISON® Q.S.E.T. Device, and associated controls/calibrators, based on the provided FDA 510(k) summary:

1. Table of Acceptance Criteria and Reported Device Performance

The FDA 510(k) summary doesn't explicitly state "acceptance criteria" as a separate section with pass/fail thresholds for each performance metric. Instead, the performance studies themselves demonstrate the device's capabilities. I've extracted key performance metrics and their reported results from the document. The implicit "acceptance criteria" here would be that these performance characteristics are adequate for the intended use and comparable to predicate devices, demonstrating substantial equivalence.

Performance MetricImplicit Acceptance Criterion (Comparison to Predicate/Clinical Utility)Reported Device Performance (LIAISON® Calprotectin Assay)
Method Comparison (vs. Commercial Calprotectin Assay)
Slope (Passing & Bablok)Close to 1.00.97 (95% CI: 0.91 to 1.00)
Intercept (Passing & Bablok)Close to 0.0 µg/g1.50 µg/g (95% CI: -2.26 to 6.46)
R-value (Linear Regression)High (close to 1.0)0.961
Positive Agreement (Borderline Elevated)High96.9% (95% CI: 92.2% - 99.1%)
Negative Agreement (Borderline Elevated)High88.9% (95% CI: 73.9% - 96.9%)
Positive Agreement (Borderline Normal)High97.8% (95% CI: 92.4% - 99.7%)
Negative Agreement (Borderline Normal)High94.4% (95% CI: 86.3% - 98.5%)
Comparative Clinical Studies (IBD vs. non-IBD)
Clinical Sensitivity (Borderline Elevated)High (for IBD diagnosis)98.0% (95% CI: 93.1 - 99.8%)
Clinical Specificity (Borderline Elevated)Adequate66.8% (95% CI: 60.4 - 76.7%) -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
Clinical Sensitivity (Borderline Normal)Adequate88.2% (95% CI: 80.4 - 93.8%)
Clinical Specificity (Borderline Normal)High (for non-IBD diagnosis)90.6% (95% CI: 84.4 - 94.9%)
Comparative Clinical Studies (IBD vs. IBS)
Clinical Sensitivity (Borderline Elevated)High (for IBD differentiation)98.0% (95% CI: 93.1 - 99.8%)
Clinical Specificity (Borderline Elevated)Adequate (for differentiating from IBS)65.7% (95% CI: 53.1 - 76.9%)
Clinical Sensitivity (Borderline Normal)Adequate88.2% (95% CI: 80.4 - 93.8%)
Clinical Specificity (Borderline Normal)High (for differentiating from IBS)88.1% (95% CI: 77.8 - 94.7%)
Analytical Measuring Range (AMR)Established range of accurate measurement5 - 800 µg/g
Limit of Blank (LoB)Low0.107 µg/g
Limit of Detection (LoD)Low0.395 µg/g
Limit of Quantitation (LoQ)Low0.400 µg/g
Accuracy/RecoveryMean recovery close to 100%Overall mean recovery: 103% (range 98-108%)
Q.S.E.T. Device Method Comparison (vs. Weigh Method)
Slope (Passing-Bablok)Close to 1.00.96 (95% CI: 0.92 to 1.02)
Correlation rHigh0.970
Overall Agreement (Borderline Elevated)High93.0% (95% CI: 87.1% - 96.7%)
Overall Agreement (Borderline Normal)High95.3% (95% CI: 90.1% - 98.3%)

2. Sample Sizes Used for the Test Set and Data Provenance

  • Method Comparison (LIAISON Calprotectin vs. Commercial Calprotectin Assay):
    • Sample Size: 164 stool samples.
    • Data Provenance: Not explicitly stated, but implies clinical samples spanning the assay range. It's retrospective in the sense that these samples were then tested on two devices.
  • Comparative Clinical Studies (IBD vs. non-IBD, IBD vs. IBS):
    • Sample Size: 240 prospectively collected human stool specimens.
    • Data Provenance: Prospectively collected from subjects with signs and symptoms suggestive of IBD or IBS. Geographic origin is not specified.
  • Expected Values (Reference Ranges):
    • Sample Size:
      • Apparently Healthy: 127 subjects (15 pediatric, 112 adult)
      • IBD: 102 subjects (19 pediatric, 333 adults - Note: The document states 19 pediatric and 333 adults for IBD/IBS subjects, but the IBD specific count is 102 total in the table, suggesting a subset. Clarification needed if this were a definitive study report.).
      • IBS: 67 subjects (subset of the 19 pediatric and 333 adults mentioned for IBD/IBS).
      • Other GI: 71 subjects.
    • Data Provenance: Stool samples from apparently healthy donors and subjects with physician-diagnosed IBS and IBD. Not explicitly stated if these were the same 240 samples from the clinical studies or an additional set.
  • LIAISON® Q.S.E.T. Device Accuracy (Method Comparison of Extraction):
    • Sample Size: 128 human stool samples.
    • Data Provenance: Not explicitly stated, but human stool samples spanning the measuring range.

3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications

  • Method Comparison (LIAISON Calprotectin vs. Commercial Calprotectin Assay): Ground truth was established by another commercial calprotectin assay, not human experts.
  • Comparative Clinical Studies (IBD vs. non-IBD, IBD vs. IBS):
    • Ground Truth Establishment: "Diagnosis of IBD, IBS, or other GI disorder was determined based on the results of colonoscopy, as well as other clinical findings. IBD diagnosis was confirmed by histological assessment of biopsy."
    • Number of Experts/Qualifications: Not specified. This typically would involve gastroenterologists, pathologists (for histological assessment), and other clinicians. The document does not provide details on the number or their specific qualifications (e.g., years of experience, board certification).
  • Expected Values: Ground truth was based on "physician diagnosed" health status (apparently healthy, IBS, IBD). Number of physicians or their qualifications are not specified.

4. Adjudication Method for the Test Set

  • Adjudication method for clinical diagnosis (ground truth): The document states that IBD diagnosis was confirmed by histological assessment of biopsy in addition to colonoscopy and other clinical findings. This implies a comprehensive clinical evaluation, but it doesn't specify an explicit adjudication process like a 2+1 or 3+1 reader consensus for borderline cases or disagreements among diagnostic methods/experts. It's a "clinical diagnosis" approach.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

No, an MRMC comparative effectiveness study involving human readers with and without AI assistance was not done. This device is an in-vitro diagnostic (IVD) assay that directly measures a biomarker, not an imaging analysis tool or a decision support system for human readers. Therefore, the concept of human readers improving with AI assistance is not applicable in this context.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done

Yes, the studies presented (method comparison, clinical sensitivity/specificity, analytical performance) represent the standalone performance of the LIAISON® Calprotectin assay. It is an automated chemiluminescent immunoassay run on the LIAISON® XL Analyzer, directly providing quantitative results without human interpretation as part of the primary diagnostic output.

7. The Type of Ground Truth Used

  • Clinical Studies:
    • Expert Consensus/Clinical Diagnosis: For IBD, IBS, and other GI disorders, the ground truth was "clinical diagnosis by colonoscopy, as well as other clinical findings," with "histological assessment of biopsy" confirming IBD. This is a form of expert consensus based on established clinical and pathological methods.
  • Method Comparison:
    • Comparator Device: The ground truth was established by a "commercial calprotectin assay."
  • Expected Values:
    • Physician Diagnosis: "Physician diagnosed" health status.

8. The Sample Size for the Training Set

The provided document describes a 510(k) premarket notification for an IVD kit, which typically involves analytical validation and clinical performance studies, not a "training set" in the context of machine learning model development. For IVDs, the kit components (reagents, calibrators, controls) are manufactured and their performance characteristics are validated extensively. The studies described are validation studies, not training studies. Therefore, there is no "training set" in the machine learning sense for this device. The development of the assay itself would have involved internal optimization and development work, but specific sample sizes for "training" are not typically reported in 510(k) submissions for such assays.

9. How the Ground Truth for the Training Set Was Established

As noted above, there is no "training set" in the machine learning sense. The ground truth for the validation studies (clinical and method comparison) are described in point 7.

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December 26, 2018

DiaSorin Inc. Sandra Zimniewicz Regulatory Affairs Specialist 1951 Northwestern Ave. Stillwater, Minnesota 55082-0285

Re: K182698

Trade/Device Name: LIAISON Calprotectin, LIAISON Calprotectin Control Set, LIAISON Calprotectin Calibration Verifiers, LIAISON Q.S.E.T. Buffer, LIAISON Q.S.E.T. Device Regulation Number: 21 CFR 866.5180 Regulation Name: Fecal calprotectin immunological test system Regulatory Class: Class II Product Code: NXO Dated: September 26, 2018 Received: September 27, 2018

Dear Sandra Zimniewicz:

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

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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 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) 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 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,

Douglas A. Jeffery -S

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

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Indications for Use

510(k) Number (if known) K182698

Device Name LIAISON® Calprotectin LIAISON® Q.S.E.T. Device

Indications for Use (Describe)

The DiaSorin LIAISON® Calprotectin assay is an in vitro diagnostic chemiluminescent immunoassay (CLIA) intended for the quantitative measurement, in human stool, of fecal calprotectin, a neutrophilic protein that is a marker of mucosal inflammation. The LIAISON® Calprotectin assay can be used 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 (IBS). Test results are to be used in conjunction with information obtained from the patients' clinical evaluation and other diagnostic procedures.

The test has to be performed on the LIAISON® XL Analyzer.

The DiaSorin LIAISON® O.S.E.T. Device (Quantitative Stool Extraction and Test) is intended for use in the preparation of human stool specimens for testing in the LIAISON® Calprotectin assay.

Type of Use (Select one or both, as applicable)
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X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) SUMMARY 5.0

SUBMITTED BY:Sandra ZimniewiczRegulatory Affairs SpecialistDiaSorin Inc.1951 Northwestern AvenueP.O. Box 285Stillwater, MN 55082-0285Phone (651) 351-5711Fax (651) 351-5669Email: sandra.zimniewicz@diasorin.com
DATE PREPARED:September 27, 2018
NAME OF DEVICE:
Trade Name:LIAISON® CalprotectinLIAISON® Calprotectin Control SetLIAISON® Calprotectin Calibration VerifiersLIAISON® Q.S.E.T. BufferLIAISON® Q.S.E.T. Device
Common Names/Descriptions:Calprotectin assay, Calprotectin controls, andCalprotectin calibration verifiers
Classification Names:Fecal calprotectin immunological test system:Class II, 21 CFR: 866.5180; Immunology (82)
Single (Specified) analyte controls (assayedand unassayed): Class I,21 CFR 862.1660; Clinical Chemistry (75)
Product Code:NXOJJX
PREDICATE DEVICES :PhiCal™ Test Fecal Calprotectin ImmunoassayDEN060001

DEVICE DESCRIPTION:

INTENDED USE:

The DiaSorin LIAISON® Calprotectin assay is an in vitro diagnostic chemiluminescent immunoassay (CLIA) intended for the quantitative measurement, in human stool, of fecal calprotectin, a neutrophilic protein that is a marker of mucosal inflammation. The LIAISON® Calprotectin assay can be used 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 (IBS). Test results are to be used in

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conjunction with information obtained from the patients' clinical evaluation and other diagnostic procedures. The test has to be performed on the LIAISON® XL Analyzer.

The DiaSorin LIAISON® Calprotectin Control Set is intended for use as assayed quality control samples to monitor the performance of the LIAISON® Calprotectin assay. The performance characteristics of the LIAISON® Calprotectin Control Set have not been established for any other assay or instrument platforms different from the LIAISON® XL Analyzer.

The DiaSorin LIAISON® Calprotectin Calibration Verifiers are assayed quality control materials intended for in vitro diagnostic use in the quantitative verification of calibration and reportable range of the LIAISON® Calprotectin assay when performed on the LIAISON® XL Analyzer.

The LIAISON® Q.S.E.T. Buffer (Quantitative Stool Extraction and Test) is intended for use in the preparation of human stool specimens for testing in designated DiaSorin stool assays.

The DiaSorin LIAISON® Q.S.E.T. (Quantitative Stool Extraction and Test) is intended for use in the preparation of human stool specimens for testing in the LIAISON® Calprotectin assay.

INDICATIONS FOR USE

Same as intended use.

KIT DESCRIPTION:

The LIAISON® Calprotectin assay is a sandwich assay that uses 2 monoclonal antibodies for capture and detection of calprotectin. The LIAISON® Calprotectin assay must be run on the LIAISON® XL Analyzer, a fully automated system with continuous loading.

Calprotectin is first extracted from human stool samples with LIAISON® Q.S.E.T. Buffer using either the weigh method or the LIAISON® Q.S.E.T. Device. The assay incubates extracted sample, calibrator, control, or calibration verifiers with assay buffer and paramagnetic particles coated with a monoclonal antibody that specifically recognizes the calprotectin heterocomplex. Following incubation, a wash cycle is performed to remove any unbound material. An isoluminol conjugated monoclonal antibody that recognizes calprotectin is then added to the reaction and incubated. The unbound conjugate is removed with a second wash step. Starter reagents are then added and a flash chemiluminescent reaction is initiated. The light signal is measured by a photomultiplier as relative light units (RLU) and is proportional to the concentration of calprotectin present in the calibrators, controls or samples.

All assay steps and incubations are performed by the LIAISON® XL Analyzer. The analyzer software automatically calculates the concentration of calprotectin in the sample. This concentration is expressed in ug/q.

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COMPARISON WITH PREDICATE

The LIAISON® Calprotectin, LIAISON® Calprotecin Controls and LIAISON® Calprotecin Calibratin Verifiers are prepackaged reagents for use on an automated clinical chemistry analyzer. The PhiCal™ Test consists of prepackaged reagents for non/semiautomated manual use. A comparison of the similarities and differences between the devices are provided in the following table:

Table 1: Table of Similarities - LIAISON® Calprotectin
CharacteristicNew DeviceDiaSorinLIAISON® CalprotectinPredicate DevicePhiCal™ TestFecal CalprotectinImmunoassay(DEN060001)
Intended UseThe DiaSorin LIAISON® Calprotectinassay is an in vitro diagnosticchemiluminescent immunoassay (CLIA)intended for the quantitativemeasurement, in human stool, of fecalcalprotectin, a neutrophilic protein that isa marker of mucosal inflammation. TheLIAISON® Calprotectin assay can beused as an aid in the diagnosis ofinflammatory bowel diseases (IBD),specifically Crohn's disease andulcerative colitis, and as an aid indifferentiation of IBD from irritable bowelsyndrome (IBS). Test results are to beused in conjunction with informationobtained from the patients' clinicalevaluation and other diagnosticprocedures. The test has to beperformed on the LIAISON® XLAnalyzer.The PhiCal™ test is aquantitative ELISA formeasuring, in human stool,concentrations of fecalcalprotectin, a neutrophilicprotein that is a marker ofmucosal inflammation. TheThe PhiCal ™ test can beused as an in vitro diagnosticaid in the diagnosis ofinflammatory bowel diseases(IBD), Crohn's disease andulcerative colitis, and todifferentiate IBD from irritablebowel syndrome.
Measured AnalyteCalprotectinSame
Assay TypeSolid phase immunoassaySame
ResultsQuantitativeSame
Sample TypeExtracted Human StoolSame
Amount ofSpecimen requiredfor ManualExtraction$50 - 100$ mg$80 - 120$ mg
LIAISON®Q.S.E.T.DeviceStoolSampling Quantity$10.5$ mg $\pm 0.1$ mgNA
Table 1 - Continued
Interpretation< 50 = Normal50 - 120 µg/g = Borderline> 120 µg/g = Elevated< 15.625 – 50 µg/g = Normal50 - 120 µg/g = Borderline> 120 µg/g = Abnormal
Kit Storage2-8°CSame

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Table 2: Table of Differences - LIAISON® Calprotectin
CharacteristicNew DeviceDiaSorinLIAISON® CalprotectinPredicate DevicePhiCal™ TestFecal CalprotectinImmunoassay (DEN060001)
Operating PrincipleAutomated ChemiluminescentImmunoassay (CLIA)Manual/Semi-automated ELISA
Solid PhaseMagnetic particles coated mousemonoclonal antibodies tocalprotectinMicrowells coated with rabbitpolyclonal antibody tocalprotectin
ConjugateMouse monoclonal antibodyagainst calprotectin conjugated toan isoluminol derivativeAlkaline phosphatase labelledrabbit IgG antibodies againstcalprotectin
AnalyticalMeasuring Range5 - 800 µg/g15.6 - 250 µg/g
ExtractionProcedureWeigh Method orLIAISON® Q.S.E.T. Device methodExtraction Buffer providedseparatelyWeigh MethodExtraction Buffer included with kit
Calibrators2 Levels, Lyophilized5 Levels, Liquid, Ready to Use
Table 3: Table of Similarities and Differences - LIAISON® Calprotectin Control Set
CharacteristicNew DeviceDiaSorinLIAISON® CalprotectinPredicate DevicePhiCal™ TestFecal CalprotectinImmunoassay(DEN060001)
Intended UseThe DiaSorin LIAISON® CalprotectinControl Set is intended for use asassayed quality control samples tomonitor the performance of theLIAISON® Calprotectin assay. Theperformance characteristics of theLIAISON® Calprotectin Control Set havenot been established for any otherassay or instrument platforms differentfrom the LIAISON® XL Analyzer.No separate intended use,controls are included as partof the kit.

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Table 3 - Continued
AntigenRecombinant CalprotectinSame
Levels2 Levels (Low and High)Same
CharacteristicsLyophilizedProvided separatelyLiquid, Ready to UseIncluded with kit
Storage2-8°CSame
Table 4: Table of Similarities and Differences - LIAISON® Calprotectin CalibrationVerfiers
CharacteristicNew DeviceDiaSorinLIAISON® CalprotectinPredicate DeviceLIAISON® XL 1,25Dihydroxyvitamin DCalibration Verifiers(K141463)
Intended UseThe DiaSorin LIAISON® CalprotectinCalibration Verifiers are assayed qualitycontrol materials intended for in vitrodiagnostic use in the quantitativeverification of calibration and reportablerange of the LIAISON® Calprotectinassay when performed on the LIAISON®XL Analyzer.Assayed quality controlmaterials intended thequantitative verification ofcalibration and reportablerange of the LIAISON® XL1,25 Dihydroxyvitamin D.
Storage2-8°CSame
Levels4 Levels, LyophilizedProvided separatelySame
Table 5: Table of Similarities and Differences - LIAISON®Q.S.E.T. Buffer
CharacteristicNew DeviceDiaSorinLIAISON® CalprotectinPredicate DevicePhiCal™ TestFecal CalprotectinImmunoassay(DEN060001)
Intended UseThe LIAISON® Q.S.E.T. Buffer(Quantitative Stool Extraction and Test)is intended for use in the preparation ofhuman stool specimens for testing indesignated DiaSorin stool assays.No separate intended use,extraction buffer is includedas part of the kit.
Storage2-8°CSame
Concentration5X (Requires dilution to 1X workingconcentration)2.5X (Requires dilution to 1Xworking concentration)
Characteristics3 bottles x 100 mL eachLiquidProvided separately, not included with kit2 bottles x 50 mL eachLiquidIncluded with kit
Stability of 1XSolution8 weeks at 2-8°C7 days at 2-8°C

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PERFORMANCE DATA:

METHOD COMPARISON

A total of 164 stool samples spanning the assay range were tested by the LIAISON® Calprotectin assay and a commercial calprotectin assay following CLSA EP09-A3. The results were analyzed by the method of Passing & Bablok, returning a slope of 0.97 (95% Cl: 0.91 to 1.00), an intercept of 1.50 µg/q (95% Cl: -2.26 to 6.46) and an R value by linear regression of 0.961

An agreement analysis was also performed relative to the respective assay cut-off values for relevant medical decision points. Positive and negative agreement were calculated with borderline results considered as normal and as elevated/abnormaand. Results are summarized in the tables below.

Comparator Calprotectin AssayConcentration
LIAISON® Calprotectin AssayConcentration< 50 µg/g(Normal)50 – 120 µg/g(Borderline)>120 µg/g(Abnormal)Total
< 50 µg/g (Normal)324036
50 - 120 µg/g (Borderline)428234
>120 µg/g (Elevated)049094
Total363692164
Borderline considered Elevated (Abnormal)95% Confidence Interval
Positive Agreement(124/128)96.9%(92.2% - 99.1%)
Negative Agreement(32/36)88.9%(73.9% - 96.9%)
Borderline considered Normal95% Confidence Interval
Positive Agreement(90/92)97.8%(92.4% - 99.7%)
Negative Agreement(68/72)94.4%(86.3% - 98.5%)

COMPARATIVE CLINICAL STUDIES:

Clinical specificity and clinical sensitivity of the LIAISON® Calprotectin assay was determined against the clinical diagnosis by testing a total of 240 prospectively collected human stool specimens from subjects with signs and symptoms suggestive of IBD or IBS. Diagnosis of IBD, IBS, or other GI disorder was determined based on the results of colonoscopy, as well as other clinical findings. IBD diagnosis was confirmed by histological assessment of biopsy. The final diagnoses of the subjects were as follows: 102 IBD (85 adult and 17 pediatric), 67 IBS (65 adult and 2 pediatric), and 71 other gastrointestinal disorders other than IBD or IBS

IBD vs non-IBD comparison:

Sensitivity and specificity were calculated for the LIAISON Calprotectin assay as an aid in the diagnosis of IBD in all subjects. Borderline results were calculated as both normal and elevated and 95% confidence intervals were reported.

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Calprotectin(µg/g)IBDNon-IBD
Normal<50295
Borderline50-1201030
Elevated>1209013
All102138
Borderline consideredElevated95% Confidence Interval
Clinical Sensitivity100/10298.0%93.1 – 99.8%
Clinical Specificity95/13866.8%60.4 – 76.7%
Borderline considered Normal95% Confidence Interval
Clinical Sensitivity90/10288.2%80.4 - 93.8%
Clinical Specificity125/13890.6%84.4 - 94.9%

IBD vs IBS comparison:

Sensitivity and specificity were calculated for the LIAISON® Calprotectin assay to aid in the differentiation of IBD and IBS. Borderline results were calculated as both normal and elevated and 95% confidence intervals were reported.

Calprotectin(µg/g)IBDIBS
Normal<50244
Borderline50-1201015
Elevated>120908
All10267
Borderline as Elevated95% Confidence Interval
Clinical Sensitivity100/10298.0%93.1 - 99.8%
Clinical Specificity44/6765.7%53.1 - 76.9%
Borderline as Normal95% Confidence Interval
Clinical Sensitivity90/10288.2%80.4 - 93.8%
Clinical Specificity59/6788.1%77.8 - 94.7%

EXPECTED VAUES:

Stool samples from apparently healthy donors (15 subjects with ages ranging from 3-21 and 112 adults) and subjects with physician diagnosed IBS and IBD (19 subjects with ages ranging from 8-21 and 333 adults) were tested with the LIAISON® Calprotectin assay. Results were evaluated using the medical decision points of 50 µg/g and 120 µg/g which corresponds to the consensus threshold for distinguishing between IBD and IBS.

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DiagnosisNumber of Subjects and percent in LIAISON® Calprotectin
Range (µg/g)
<50.0Normal50.0-120Borderline>120ElevatedTotal
Apparently Healthy112 (88.2%)15 (11.8%)0 (0%)127 (100%)
IBD2 (2.0 %)10 (9.8%)90 (88.2%)102 (100%)
IBS44 (65.7%)15 (22.4%)8 (11.9%)67 (100%)
OTHER GI51 (71.8%)15 (21.1%)5 (7.1%)71 (100%)

Test results are to be used in conjunction with information obtained from the patients' clinical evaluation and other diagnostic procedures.

PRECISION/REPRODUCIBILITY:

A twelve (12) day study was conducted at DiaSorin Inc. A panel of six (6) human stool samples containing concentrations of analyte prepared to span the measuring range of the assay were extracted by each of three (3) operators using the manual extraction method each day over 12 operating days and tested in two (2) replicates in two (2) runs per day using one (1) lot of LIAISON® Calprotectin reagent. CLSI quideline EP5-A3 was consulted in the preparation of the testing protocol.

Sample IDnMeanµg/gWithin RunBetweenRunBetween DayBetweenOperatorTotal
SD%CVSD%CVSD%CVSD%CVSD%CV
Sample #114424.90.743.0%0.662.6%2.9111.7%1.044.2%3.2513.0%
Sample #214439.90.882.2%0.761.9%3.448.6%0.000.0%3.639.1%
Sample #31441553.992.6%3.472.2%11.267.2%8.275.3%14.949.6%
Sample #41442536.972.8%8.183.2%20.178.0%2.871.1%23.039.1%
Sample #514421.60.673.1%0.371.7%2.9313.5%0.622.9%3.0914.3%
Sample #614463915.862.5%25.314.0%39.216.1%21.033.3%53.598.4%

A five (5) day study was conducted at two (2) external laboratories and at DiaSorin Inc. A panel of six (6) human stool samples containing concentrations of analyte prepared to span the measuring range of the assay were extracted each day at each site using the manual extraction method. Kit controls were also included in the study. Each site utilized at least two (2) operators to perform the extraction and testing. The samples were tested in six (6) replicates in one (1) run each day using one (1) lot of LIAISON® Calprotectin reagent. CLSI document EP15-A3 was consulted in the preparation of the testing protocol.

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Sample IDMeanµg/gWithin RunBetween DayWithin SiteSite to SiteTotal
SD%CVSD%CVSD%CVSD%CVSD%CV
Control 151.21.192.3%3.677.2%3.837.5%0.150.3%3.837.5%
Control 22547.593.0%26.1110.3%27.0210.6%0.000.0%25.9410.2%
Sample #125.90.682.6%2.168.3%2.258.7%0.000.0%2.178.4%
Sample #242.10.781.8%5.2612.5%5.3112.6%1.874.4%5.6313.4%
Sample #31734.652.7%20.4711.9%20.9112.1%20.5511.9%29.3217.0%
Sample #42815.622.0%26.929.6%27.419.7%40.1014.3%48.5717.3%
Sample #523.40.602.6%2.6211.2%2.6811.5%0.0000.0%2.4510.5%
Sample #669518.852.7%66.069.5%68.269.8%39.615.7%78.9211.3%

LIAISON® Q.S.E.T. Device Reproducibility

To study precision of the Calprotectin dose using the Q.S.E.T. device to extract the stool sample, eight (8) stool samples spanning the analytical measuring range (AMR) were extracted using the Q.S.E.T. Device and tested once (1) per day using five (5) replicates over five (5) days by three (3) operators, for a total of 75 measurements per Each Q.S.E.T. device extraction was performed daily by each operator samples. independently. The precision study was based on CLSI EP05-A3.

Sample IDMeanµg/gRepeatabilityBetween DayWithinOperatorBetweenOperatorTotal
SD%CVSD%CVSD%CVSD%CVSD%CV
121.90.6362.9%3.17914.5%3.23214.8%0.9454.3%3.05213.9%
226.20.8073.1%1.9517.4%2.0857.9%1.8437.0%2.64310.1%
338.61.4283.7%4.72712.2%4.90312.7%1.3353.5%4.62112.0%
41666.1373.7%9.1105.5%10.696.5%8.1414.9%12.817.7%
529711.293.8%28.629.6%30.4210.2%6.5942.2%28.379.6%
650518.593.7%59.3911.8%61.7712.2%34.356.8%65.4913.0%
7238162.562.6%410.417.2%414.417.4%254.710.7%450.518.9%
84140126.83.1%309.27.5%330.18.0%228.95.5%377.19.1%

TRACEABILITY/STANDARDIZATION

The LIAISON® Calprotectin calibrator concentrations (µg/g) are referenced to an inhouse standard preparation made from recombinant calprotectin.

Each Reagent Integral requires calibration. Calibration is performed using the two (2) calibrators included with each LIAISON® Calprotectin kit. The two point kit calibrators are used to establish specific working curves based on 10 point assay master curves stored on the analyzer.

LINEARITY

Linearity is demostrated for the analytical measuring range of 5 - 800 µg/g.

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ACCURACY/RECOVERY

The mean recovery results of 5 samples tested ranged from 98 to 108% with an overall mean recovery of 103%.

SamplesDefinedConcentration(µg/g)ExpectedConcentrationn (µg/g)ObservedConcentration(µg/g)%Recovery
High Sample 1 (HS1)493
2 HS1 : 1 LS1399405101%
1 HS1 : 1 LS1351358102%
1 HS1 : 2 LS1303320106%
Low Sample 1 (LS1)209
High Sample 2 (HS2)599
2 HS2 : 1 LS2410418102%
1 HS2 : 1 LS2313330106%
1 HS2 : 2 LS2216226105%
Low Sample 2 (LS2)26.8
High Sample 3 (HS3)408
2 HS3 : 1 LS3281292104%
1 HS3 : 1 LS3215217101%
1 HS3 : 2 LS314914698%
Low Sample 3 (LS3)21.6
High Sample 4 (HS4)724
2 HS4 : 1 LS4523537103%
1 HS4 : 1 LS4420435104%
1 HS4 : 2 LS4317344108%
Low Sample 4 (LS4)117
High Sample 5 (HS5)413
2 HS5 : 1 LS5308315102%
1 HS5 : 1 LS5253271107%
1 HS5 : 2 LS5199199100%
Low Sample 5 (LS5)93.6
Mean Recovery103%

ANALYTICAL SENSITIVITY

Limit of Blank (LoB) is 0.107 µg/g.

Limit of Detection (LoD) is 0.395 µg/g.

Limit of Quantitation (LoQ) is 0.400 µg/g.

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INTERFERING SUBSTANCES

Controlled studies of potentially interfering substances and microorganisms performed in a human stool sample extract at a calprotectin level of approximately 50 µg/g showed no interference (< 10% change in test results) in the LIAISON® Calprotectin assay at the highest concentration for each substance or microorganisms listed below. The testing was based on CLSI-EP7-A2.

Drug/SubstanceConcentrationTested
Stearic acid2.65 mg/mL
Palmitic acid1.3 mg/mL
Hemoglobin6.7 µg/mL
S100A12 protein21.0 µg/mL
Barium sulfate5.0 mg/mL
Imodium AD®6.67 µg/mL
Kaopectate®0.87 mg/mL
Metronidazole12.5 mg/mL
Mucin3.33 mg/mL
Mylanta®4.2 mg/mL
Pepto Bismol®0.87 mg/mL
Polyethylene glycol 335079.05 mg/mL
Prilosec®0.5 mg/mL
Simethicone0.625 mg/mL
Tagamet®0.5 mg/mL
Tums®0.5 mg/mL
Vancomycin hydrochloride2.5 mg/mL
Mesalamine5.0 mg/mL
Prednisone0.3 mg/mL
Lansoprazole0.2 mg/mL
Sulfamethoxazole1.6 mg/mL
Ciprofloxacin1.25 mg/mL
Vitamin E0.3 mg/mL
Vitamin D30.1 µg/mL
Provitamin A5 mg/mL
Azathioprine0.2 mg/mL
Vitamin C0.1 mg/mL
MicroorganismFinal concentrationof variant in sample
Citrobacter freundii1.2 x 108 CFU/mL
Escherichia coli1.2 x 108 CFU/mL
Klebsiella pneumonia1.2 x 108 CFU/mL
Salmonella enterica1.2 x 108 CFU/mL
Shigella boydii1.2 x 108 CFU/mL
Yersinia enterolitica1.2 x 108 CFU/mL

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LIAISON® Q.S.E.T. DEVICE ACCURACY

One hundred twenty-eight (128) human stool samples spanning the measuring range of the LIAISON® Calprotectin assay were extracted using the LIAISON® Q.S.E.T. Device and the weigh method to determine if these extraction methods provide equivalent results. Each human stool extract was tested in singlicate in the same run using one (1) LIAISON® Calprotectin assay reagent lot. The results were analyzed by Passing-Bablok regression analysis of manual weigh method versus device method.

y- intercept(95% CI)Slope(95% CI)Bias at 50 µg/g(95% CI)Bias at 120 µg/g(95% CI)Correlation r
-1.12 µg/g(-2.81 to 0.60)0.96(0.92 to 1.02)-2.914 µg/g(-5.154 to -0.529)-5.428 µg/g(-9.575 to -0.450)0.970

Percent agreement analysis performed between the two methods of extraction obtained the following results.

Manual Extraction (weight)
LIAISON® Q.S.E.T.DeviceElevatedBorderlineNormalTotal
Elevated360036
Borderline623332
Normal075360
Total423056128
Borderline considered Elevated95% Confidence IntervalBorderline considered Normal95% Confidence Interval
Positive Agreement(65/72)90.3%(81.0% - 96.0%)Positive Agreement(36/42)85.7%(71.5% - 94.6%)
Negative Agreement(53/56)94.6%(85.1% - 99.0%)Negative Agreement(86/86)100%(95.8% - 100.0%)
Overall(119/128)93.0%(87.1% - 96.7%)Overall(122/128)95.3%(90.1% - 98.3%)

LIAISON® Q.S.E.T. SAMPLE COLLECTION REPRODUCIBILITY

LIAISON® Q.S.E.T. Device weight reproducibility was tested using seven (7) human stool samples ranging from 2 - 6 on the Bristol Stool Form Scale (BSFS) were sampled by three (3) operators with five (5) replicates per sample per operator. Samples on the Q.S.E.T. device were weighed against the empty Q.S.E.T device.

Repeatability of the sample collection weight was assessed across all seven (7) samples, replicates and operators.

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Mean Sample Weight10.5 mg
Median Sample Weight10.5 mg
Range9.0-12.0 mg
95% CI10.4-10.6 mg
SD0.511 mg
%CV4.9%

Sample collection performance of LIAISON® Q.S.E.T. Device

REAGENT STABILITY

LIAISON® CalprotectinStability
Calibration Curve28 days
Reagent Integral Open Use storage On-board Analyzer56 days
Reagent Integral Open Use storage at 2-8°C56 days
Reconstituted Calibrator open use at 18-25°C (room temperature)6 hours
Reconstituted Calibrator open use at 2 - 8°C28 days
LIAISON® Calprotectin Control SetStability
Reconstituted Calibrator open use at 18-25°C (roomtemperature)6 hours
Reconstituted Controls open use at 2 - 8°C28 days
Reconstituted Controls Open Use at -20°C8 weeks
Freeze/Thaw4 cycles
LIAISON® Calprotectin Calibration VerifiersStability
Reconstituted Calibrator open use at 18-25°C (roomtemperature)6 hours
Reconstituted Controls open use at 2 - 8°C28 days
Reconstituted Controls Open Use at -20°C8 weeks
Freeze/Thaw4 cycles
LIAISON® Q.S.E.T. BufferStability
Diluted (1X) at 2 - 8°C8 weeks

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SPECIMEN STABILITY

Studies were performed to determine the stability of sample at different storage conditions. The results are provided in the tables below.

Raw Stool Specimen
Storage ConditionStability
Refrigerated at 2- 8°C72 hours
Store samples frozen at -20°C if not tested within 72 hours.
Freeze/Thaw cycles3 cycles
Sample Extract - Weigh Method
Storage ConditionStability
Room temperature (18 - 25°C)8 hours
Refrigerated at 2- 8°C72 hours
Sample Extract - LIAISON® Q.S.E.T. Device Method
Storage ConditionStability
Room temperature (18 - 25°C)4 hours
Refrigerated at 2- 8°C (without centrifugation)6 hours
Refrigerated at 2- 8°C (with centrifugation)7 days

CONCLUSION:

The material submitted in this premarket notification is complete and supports a substantial equivalence decision. The labeling is sufficient and it satisfies the requirements of 21CFR 809.10.

§ 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).