EliA Celikey IgG Immunoassay; EliA GliadinDP IgA Immunoassay; EliA GliadinDP IgG Immunoassay

K181871 · Phadia AB · MVM · Mar 1, 2019 · Immunology

Device Facts

• Record ID: K181871
• Device Name: EliA Celikey IgG Immunoassay; EliA GliadinDP IgA Immunoassay; EliA GliadinDP IgG Immunoassay
• Applicant: Phadia AB
• Product Code: MVM · Immunology
• Decision Date: Mar 1, 2019
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5660
• Device Class: Class 2

Intended Use

EliA Celikey IgG is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to tissue transglutaminase (tTG) in human serum and EDTA-plasma. EliA Celikey IgG is based on recombinant human tissue transglutaminase as antigen and is useful as an aid in the clinical diagnosis of patients with celiac disease in conjunction with other laboratory and clinical findings. EliA Celikey IgG uses the EliA IgG method on the instrument Phadia 2500/5000. EliA GliadinDP IgA is intended for the in vitro semi-quantitative measurement of IgA antibodies directed to gliadin in human serum or plasma (Li-heparin, EDTA) to aid in the diagnosis of celiac disease in conjunction with other laboratory and clinical findings. EliA GliadinDP IgA uses the EliA IgA method on the instrument Phadia 2500/5000. EliA GliadinDP IgG is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to gliadin in human serum or plasma (Li-heparin, EDTA) to aid in the diagnosis of celiac disease in conjunction with other laboratory and clinical findings. EliA GliadinDP IgG uses the EliA IgG method on the instrument Phadia 2500/5000.

Device Story

Automated semi-quantitative solid-phase fluoroimmunoassays for detection of celiac disease-associated autoantibodies (tTG IgG, gliadin IgA/IgG). Input: human serum or plasma samples. Process: samples incubated in polystyrene wells coated with recombinant tTG or synthetic deamidated gliadin peptides; bound antibodies detected via enzyme-labeled anti-IgG or anti-IgA conjugate; fluorescence measured after development/stop solution addition. Operation: performed on Phadia 2500/5000 instruments; random access; automated pipetting using disposable tips. Output: fluorescence response compared to calibrators to determine antibody concentration. Clinical use: aids diagnosis of celiac disease; results interpreted by clinicians alongside other laboratory/clinical findings.

Clinical Evidence

Bench testing only. Precision, linearity, LoD/LoQ, and instrument comparison studies performed. Instrument comparison (n=100-105 samples per assay) showed high agreement with predicate (R > 0.99). Reference range established using 400 healthy donors. Clinical performance data was previously reviewed in K062583 and K093459.

Technological Characteristics

Polystyrene wells coated with recombinant human tTG or synthetic deamidated gliadin peptides. Detection via beta-galactosidase labeled anti-IgG/IgA conjugate. Fluorescence-based measurement. Automated system (Phadia 2500/5000) with disposable pipette tips. Standards: CLSI EP05-A3, EP6-A, EP17-A2.

Indications for Use

Indicated for the in vitro semi-quantitative measurement of IgG antibodies to tissue transglutaminase (tTG) or IgA/IgG antibodies to gliadin in human serum or plasma to aid in the diagnosis of celiac disease in conjunction with other clinical findings.

Regulatory Classification

Identification

A multiple autoantibodies immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoantibodies (antibodies produced against the body's own tissues) in serum and other body fluids. Measurement of multiple autoantibodies aids in the diagnosis of autoimmune disorders (disease produced when the body's own tissues are injured by autoantibodies).

Predicate Devices

• EliA Celikey IgG on Phadia 250 instrument (K062583)
• EliA GliadinDP IgA on Phadia 250 instrument (K093459)
• EliA GliadinDP IgG on Phadia 250 instrument (K093459)

Submission Summary (Full Text)

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March 1, 2019

Phadia AB % Sheryl Skinner Regulatory Director, RA/QA Phadia US Inc. 4169 Commercial Avenue Portage, Michigan 49002

Re: K181871

Trade/Device Name: EliA Celikey IgG Immunoassay; EliA GliadinDP IgA Immunoassay; EliA GliadinDP IgG Immunoassay Regulation Number: 21 CFR 866.5660 Regulation Name: Multiple autoantibodies immunological test system Regulatory Class: Class II Product Code: MVM, MST Dated: July 10, 2018 Received: July 12, 2018

Dear Sheryl Skinner:

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.

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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 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 ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Douglas A. Jeffery -S

Doug Jeffery, Ph.D. Branch Chief Immunology and Flow Cytometry Branch 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) K181871

#### Device Name

EliA Celikey IgG Immunoassay, EliA GliadinDP IgA Immunoassay, EliA GliadinDP IgG Immunoassay

#### Indications for Use (Describe)

EliA Celikey IgG is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to tissue transglutaminase (tTG) in human serum and EDTA-plasma. EliA Celikey IgG is based on recombinant human tissue transglutaminase as antigen and is useful as an aid in the clinical diagnosis of patients with celiac disease in conjunction with other laboratory and clinical findings. EliA Celikey IgG uses the EliA IgG method on the instrument Pladia 2500/5000.

EliA GliadinDP IgA is intended for the in vitro semi-quantitative measurement of IgA antibodies directed to gliadin in human serum or plasma (Li-heparin, EDTA) to aid in the diagnosis of celiac disease in conjunction with other laboratory and clinical findings. EliA GliadinDP IgA uses the EliA IgA method on the instrument Phadia 2500/5000.

EliA GliadinDP IgG is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to gliadin in human serum or plasma (Li-heparin, EDTA) to aid in the diagnosis of celiac disease in conjunction with other laboratory and clinical findings. EliA GliadinDP IgG uses the EliA IgG method on the instrument Phadia 2500/5000.

Type of Use (Select one or both, as applicable)

| <div> <span> <span style="font-size: 16px;">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) </span> </div> |
|--------------------------------------------------------------------------------------------------------------------------|
| <div> <span> <span style="font-size: 16px;">☐</span> Over-The-Counter Use (21 CFR 801 Subpart C) </span> </div> |

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# A.6 510(k) Summary of Safety and Effectiveness per 21CFR 807.92(c).

This summary of safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR Part 807.92.

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY AND INSTRUMENT COMBINATION TEMPLATE

## A. 510(k) Number: K181871

## B. Purpose for Submission:

Adding previously cleared assays on a new instrument platform (Phadia® 2500/5000)

#### C. Measurands:

Anti-tissue transglutaminase (tTG) IgG antibody Anti-gliadin IgA and anti-gliadin IgG autoantibodies

#### D. Type of Test:

Semi-quantitative measurement immunoassays

## E. Applicant:

Phadia AB Rapsgatan 7P P.O. Box 6460 SE-751 37 Uppsala, Sweden Tel: [phone]

510(k) Contact Person: Sheryl Skinner Associate Director, RA/QA ImmunoDiagnostics US Thermo Fisher Scientific Phadia US Inc. 4169 Commercial Avenue Portage, Mi 49002, USA Tel. [phone] [e-mail]

Date of Summary Preparation:

February 28, 2019

## F. Proprietary and Established Names:

EliA Celikey IgG Immunoassay EliA Gliadin®º IgA Immunoassay EliA Gliadin®º IgG Immunoassay

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#### G. Regulatory Information:

- 1. Requlation section:
- 21 CFR §866.5660 Multiple autoantibodies immunological test system 21 CFR §866.5750 – Radioallergosorbent (RAST) Immunological Test System
- 2. Classification:

Class II

- 3. Product code:
MVM Autoantibodies, endomysial (tissue transqlutaminase) autoantibodies, gliadin MST

- 4. Panel: Immunology (82)
## H. Intended use(s):

1. Intended use(s):

EliA Celikey IqG is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to tissue transglutaminase (tTG) in human serum and EDTAplasma. EliA Celikey IgG is based on recombinant human tissue transqlutaminase as antigen and is useful as an aid in the clinical diagnosis of patients with celiac disease in conjunction with other laboratory and clinical findings. EliA Celikey IqG uses the EliA IgG method on the instrument Phadia 2500/5000.

EliA Gliadin®° IgA is intended for the in vitro semi-quantitative measurement of IgA antibodies directed to gliadin in human serum or plasma (Li-heparin, EDTA) to aid in the diagnosis of celiac disease in conjunction with other laboratory and clinical findings. EliA Gliadin® IgA uses the EliA IgA method on the instrument Phadia 2500/5000.

EliA Gliadin®P IgG is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to gliadin in human serum or plasma (Li-heparin, EDTA) to aid in the diagnosis of celiac disease in conjunction with other laboratory and clinical findings. EliA Gliadin® IgG uses the EliA IgG method on the instrument Phadia 2500/5000.

2. Indication(s) for use: Same as intended use

3. Special conditions for use statement(s):

For prescription use only

- Special instrument requirements: ধ
Performance studies were obtained from the Phadia® 2500/5000 instrument This device is not for point-of-care use.

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#### . Device Description:

The method-specific reagents are identical with K062583 (EliA Celikey IgG) and K093459 (EliA Gliadin® IgA and EliA Gliadin® IgG), but are filled in containers specific for the Phadia 2500/5000 instrument. Each device consists of: Test Wells:

- EliA Celikey IgG Wells are coated with human recombinant tissue transglutaminase (tTG) - 2 carriers (12 wells each), ready to use;
- EliA Gliadin®° IgA Wells are coated with synthetic deamidated gliadin peptides -- 4 carriers (12 wells each), ready to use;
- -EliA Gliadin®° IgG Wells are coated with synthetic deamidated gliadin peptides - 4 carriers (12 wells each), ready to use;

EliA Sample Diluent:

- EliA Sample Diluent: PBS containing BSA, detergent and 0.095% (w/v) sodium azide - 6 bottles, 48 mL each, ready to use; or 6 bottles, 400 mL each, ready to use;
EliA IgG reagents (required for EliA Celikey IgG and EliA Gliadin®º IgG):

- EliA IgG Coniugate 50 or 200: ß-Galactosidase labeled anti-laG (mouse monoclonal antibodies) in PBS containing BSA and 0.06% (w/v) sodium azide – 6 wedge shaped bottles, 5 mL each, ready to use; or 6 wedge shaped bottles, 19 mL each, ready to use
- EliA IgG Calibrator Strips: Human IgG (0, 4, 10, 20, 100, 600 µg/L) in PBS containing BSA, detergent and 0.095% (w/v) sodium azide - 5 strips, 6 singleuse vials per strip, 0.3 mL each, ready to use;
- EliA IgG Curve Control Strips: Human IgG (20 µg/L) in PBS containing BSA, detergent and 0.095% (w/v) sodium azide – 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use;
- EliA IgG Calibrator Well: Coated with mouse monoclonal antibodies 4 carriers (12 wells each), ready to use.

EliA IgA reagents (required for EliA Gliadin®º IgA):

- EliA IgA Conjugate 50 or 200: ß-Galactosidase labeled anti-IgA (mouse monoclonal antibodies) in PBS containing BSA and 0.06% (w/v) sodium azide -6 wedge shaped bottles, 5 mL each, ready to use; or 6 wedge shaped bottles, 19 mL each, ready to use
- -EliA IgA Calibrator Strips: Human IgA (0, 0.3, 1.5, 5, 15, 80 µg/L) in PBS containing BSA, detergent and 0.095% (w/v) sodium azide - 5 strips, 6 singleuse vials per strip, 0.3 mL each, ready to use;
- -EliA IgA Curve Control Strips: Human IgA (20 µg/L) in PBS containing BSA, detergent and 0.095% (w/v) sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use;
- -EliA IgA Calibrator Well: Coated with mouse monoclonal antibodies - 4 carriers (12 wells each), ready to use.

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The Phadia EliA Immunodiagnostic System is an automated system for immunodiagnostic testing. The EliA reagents are available as modular packages, each purchased separately. All packages are required to carry out EliA Celikey IgG and EliA Gliadin®º IgA and EliA Gliadinºº IgG tests.

#### J. Substantial Equivalence Information:

1. Predicate device name(s) and 510(k) number(s): EliA Celikey IgG on Phadia 250 instrument, K062583 EliA Gliadin®P IgA on Phadia 250 instrument, K093459 EliA Gliadin®° IgG on Phadia 250 instrument, K093459

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#### 2. Comparison with predicate device:

#### EliA Celiac Immunoassays on Phadia 250 and Phadia 2500/5000 instruments – Similarities to predicate devices

| Feature | Predicate Device<br>Phadia 250 | New Device<br>Phadia 2500/5000 |
|--------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use<br>EliA Celikey IgG | EliA Celikey IgG is intended for<br>the in vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to tissue<br>transglutaminase (tTG) in human<br>serum and plasma. EliA Celikey<br>IgG is based on recombinant<br>human tissue transglutaminase<br>as antigen and is useful as an<br>aid in the clinical diagnosis of<br>patients with Celiac disease. EliA<br>Celikey IgG uses the EliA IgG<br>method on the instrument<br>ImmunoCAP 250*.<br>(* Former name of Phadia 250) | EliA Celikey IgG is intended for<br>the in vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to tissue<br>transglutaminase (tTG) in human<br>serum and EDTA-plasma. EliA<br>Celikey IgG is based on<br>recombinant human tissue<br>transglutaminase as antigen and<br>is useful as an aid in the clinical<br>diagnosis of patients with celiac<br>disease in conjunction with other<br>laboratory and clinical findings.<br>EliA Celikey IgG uses the EliA<br>IgG method on the instrument<br>Phadia 2500/5000. |
| Intended Use<br>EliA GliadinDP IgA | EliA GliadinDP IgA is intended for<br>the in vitro semi-quantitative<br>measurement of IgA antibodies<br>directed to gliadin in human<br>serum and plasma (heparin,<br>EDTA, citrate) to aid in the<br>diagnosis of celiac disease in<br>conjunction with other laboratory<br>and clinical findings. EliA<br>GliadinDP IgA uses the EliA IgA<br>method on the instrument<br>Phadia 250. | EliA GliadinDP IgA is intended for<br>the in vitro semi-quantitative<br>measurement of IgA antibodies<br>directed to gliadin in human<br>serum or plasma (Li-heparin,<br>EDTA) to aid in the diagnosis of<br>celiac disease in conjunction<br>with other laboratory and clinical<br>findings. EliA GliadinDP IgA uses<br>the EliA IgA method on the<br>instrument Phadia 2500/5000. |
| Intended Use<br>EliA GliadinDP IgG | EliA GliadinDP IgG is intended for<br>the in vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to gliadin in human<br>serum and plasma (heparin,<br>EDTA, citrate) to aid in the<br>diagnosis of celiac disease in<br>conjunction with other laboratory<br>and clinical findings. EliA<br>GliadinDP IgG uses the EliA IgG<br>method on the instrument<br>Phadia 250. | EliA GliadinDP IgG is intended for<br>the in vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to gliadin in human<br>serum or plasma (Li-heparin,<br>EDTA) to aid in the diagnosis of<br>celiac disease in conjunction<br>with other laboratory and clinical<br>findings. EliA GliadinDP IgG uses<br>the EliA IgG method on the<br>instrument Phadia 2500/5000. |
| Analytical<br>technology:<br>Immuno-<br>fluorescence<br>measurement | Same | Same |
| Assay process | Same | Same |
| Common, dedicated<br>Phadia reagents | Same | Same<br>Introduction of new article<br>numbers for Development<br>Solution, Stop Solution and<br>Washing Solution is only due to<br>larger filling volumes which are<br>required for the bigger<br>instruments Phadia 2500/5000 |
| Result calculation<br>software; Phadia<br>Information Data<br>Manager (IDM) | Same | Same |
| Sample volume | 90 µL (20 µL of non-diluted<br>sample) | 90 µL (20 µL of non-diluted<br>sample) |
| Incubation<br>temperature | 37°C | 37°C |
| Conjugate volume | 90 µL | 90 µL |
| Development<br>Solution Volume | 90 µL | 90 µL |
| Stop Solution<br>Volume | 200 µL | 200 µL |
| Assay set-up | Random access | Random access |
| Reagent packaging<br>size | Various/Common | Various/Common<br>Introduction of new article<br>number for EliA Sample Diluent<br>(83-1071-01) is only due to<br>larger filling volume. |
| Onboard storage of<br>reagents | Yes | Yes |
| Feature | Predicate Device<br>Phadia 250 | New Device<br>Phadia 2500/5000 |
| Sample matrix;<br>Serum or plasma<br>type as indicated in<br>the DFU dependent<br>on assay | For EliA Celikey IgG: Serum or<br>plasma (EDTA, citrate)<br>For EliA GliadinDP IgA and IgG:<br>Serum or plasma (heparin,<br>EDTA, citrate) | For EliA Celikey IgG: serum or<br>EDTA-plasma<br>For EliA GliadinDP IgA and IgG:<br>Serum or plasma (Li-heparin,<br>EDTA) |
| Daily throughput | ~250 tests | ~2500/5000 tests |
| Sample Dilution | Phadia 250 uses a steel pipette<br>to dilute the samples in Dilution<br>Plates (Art.No. 12-3907-08) | Phadia 2500/5000 uses<br>disposable Pipette Tips in Racks<br>(Art No. 12-3805-04) for<br>pipetting samples in Dilution<br>Well (Art.No. 12-4005-69) |
| Risk for carry-over | The warning “DO NOT REUSE”<br>in the Phadia 250 DFU for EliA<br>Conjugates is due to the fact that<br>a low risk of conjugate<br>contamination by carry-over from<br>samples was identified. In order<br>to reduce the risk, the single use<br>statement for the conjugate was<br>included in the Phadia 250 DFU. | When running EliA tests on the<br>Phadia 2500/5000 instruments,<br>there is no need for this warning<br>statement because these<br>instruments use disposable tips<br>for pipetting samples and a<br>separate pipette for the<br>conjugate, and carry-over from<br>samples to conjugate is<br>impossible. |
| Loading of EliA<br>Carriers | EliA carriers are loaded manually<br>on the Loading Tray from where<br>they can be processed directly or<br>transferred to the cooled storage<br>compartment. | The Phadia 2500/5000<br>instruments do not have such a<br>Loading Tray. The EliA carriers<br>are loaded into racks which are<br>directly transferred to the cooled<br>storage compartment |
| Barcode reader | The Phadia 250 instrument has<br>a built-in barcode reader at the<br>front of the instrument, but the<br>operator needs to scan the<br>barcodes manually by showing<br>the reagents to the barcode<br>reader. Alternatively, the<br>operator can also enter the<br>characters below the barcode<br>manually. | The Phadia 2500/5000<br>instruments dispose of a built-in<br>barcode reader, and the<br>reagents are on a moving belt<br>which conveys them past the<br>barcode reader. The lot-specific<br>information will be read<br>automatically by the instrument<br>during loading. |
| Process time / Time<br>to patient result | Phadia 250 needs 1 minute to<br>process one Well.<br>Phadia 250 provides the results<br>at a one minute interval. | Phadia 2500/5000 instruments<br>process two Wells in parallel in<br>48 seconds.<br>Phadia 2500/5000 provides the<br>results at a 24 seconds interval. |

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# EliA Celiac Immunoassays on Phadia 250 and Phadia 2500/5000 instruments – Differences to predicate device

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#### K. Standard/Guidance Document Referenced (if applicable):

CLSI EP05-A3; Evaluation of Precision Performance of Quantitative Measurement Methods: September 2014

CLSI EP06-A Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach: April 2003

CLSI EP17-A, Protocols for Determination of Limits of Detection and Limits of Quantification: October 2004.

CLSI EP09-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples

#### L. Test Principle:

The EliA wells are molded cups comparable to excised wells from a microtiter plate. They are made of polystyrene and are coated with the respective antigen. The wells are at the same time a holder of the coupled antigen for convenient automation and a reaction chamber with reaction/washing solution handling based on pipetting to add and aspiration to remove liquids.

The EliA wells are coated with human recombinant tissue transglutaminase, or with synthetic deamidated gliadin peptides. If present in the patient's specimen, antibodies to these proteins bind to the specific antigen. After washing away nonbound antibodies, enzyme-labeled antibodies against human IgA or IgG antibodies (EliA IgA Conjugate or EliA IgG Conjugate) are added to form an antibody-conjugate complex. After incubation, non-bound conjugate is washed away and the bound complex is incubated with a Development Solution. After stopping the reaction, the fluorescence in the reaction mixture is measured. The higher the response value, the more specific IgA or IgG is present in the specimen. To evaluate test results, the response for patient samples is compared directly to the response for calibrators.

#### M. Performance Characteristics (if/when applicable):

1. Analytical performance:

- a. Precision/Reproducibility:
To determine the precision of the assay, the variability was assessed in a study with a total of 21 runs (3 instruments x 7 runs).

The study was performed with 1 run/dav over a period of 7 days. Each sample was tested in four replicates/run giving in total 84 replicates per sample. The data was calculated against the calibration curve from Day 1.

The results are summarized in the table below:

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| Mean<br>(EliA U/mL) | Within-Run | | Between-Run | | Between-Instrument | | Total Imprecision | |
|---------------------|------------|------|-------------|-----|--------------------|------|-------------------|------|
| | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 1.6 | 0.2 | 10.0 | 0.1 | 7.2 | 0.4 | 25.0 | 0.4 | 27.9 |
| 7.6 | 0.2 | 2.8 | 0.2 | 3.1 | 0.3 | 4.2 | 0.4 | 5.9 |
| 9.6 | 0.3 | 2.7 | 0.3 | 2.6 | 0.5 | 5.4 | 0.6 | 6.6 |
| 104.4 | 2.7 | 2.6 | 3.1 | 3.0 | 3.4 | 3.3 | 5.3 | 5.1 |
| 274.6 | 8.4 | 3.1 | 6.9 | 2.5 | 9.5 | 3.5 | 14.5 | 5.3 |

## EliA Celikey IgG on Phadia 2500/5000

#### EliA Gliadin®º IgA on Phadia 2500/5000

| Mean<br>(EliA<br>U/mL) | Within-<br>Run | Between-<br>Run | | Between-<br>Instrument | | Total<br>Imprecision | | |
|------------------------|----------------|-----------------|-----|------------------------|------|----------------------|------|------|
| | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 0.8 | 0.1 | 8.9 | 0.0 | 5.7 | 0.1 | 14.9 | 0.1 | 18.2 |
| 7.4 | 0.2 | 3.0 | 0.1 | 1.5 | 0.1 | 1.3 | 0.3 | 3.6 |
| 8.7 | 0.2 | 2.8 | 0.2 | 2.6 | 0.2 | 2.3 | 0.4 | 4.5 |
| 42.8 | 1.5 | 3.6 | 0.9 | 2.1 | 1.2 | 2.8 | 2.1 | 5.0 |
| 135.3 | 5.8 | 4.3 | 3.8 | 2.8 | 10.4 | 7.7 | 12.5 | 9.3 |

#### EliA Gliadin®º IgG on Phadia 2500/5000

| Mean | | Within-<br>Run | Between-<br>Run | | Between-<br>Instrument | | Total<br>Imprecision | |
|----------------|-----|----------------|-----------------|-----|------------------------|-----|----------------------|------|
| (EliA<br>U/mL) | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 3.6 | 0.3 | 8.2 | 0.2 | 5.2 | 0.3 | 8.6 | 0.5 | 13.0 |
| 7.2 | 0.2 | 3.3 | 0.4 | 5.4 | 0.2 | 3.1 | 0.5 | 7.0 |
| 9.3 | 0.3 | 2.8 | 0.5 | 5.2 | 0.0 | 0.0 | 0.5 | 5.9 |
| 73.7 | 2.6 | 3.5 | 4.9 | 6.7 | 2.2 | 3.0 | 6.0 | 8.1 |
| 219.6 | 8.2 | 3.7 | 13.2 | 6.0 | 6.8 | 3.1 | 17.0 | 7.7 |

#### b. Linearity/assay reportable range:

Four patient serum samples (five for Celikey IgG) were diluted in sample diluent and tested with one batch of EliA Celikey IgG, EliA Gliadin®° IgA and EliA Gliadin®° IgG Immunoassays and one set of system reagents on Phadia 2500/5000. The results of the regression analysis are summarized below:

| Dilution range<br>(EliA U/mL) | Slope | Intercept | R2 |
|-------------------------------|-------|-----------|------|
| <b>1.3</b> – 35.5 | 1.03 | 0.49 | 0.99 |
| 1.8 – 32.9 | 1.02 | 0.69 | 0.99 |
| 2.2 – 196.6 | 1.02 | 1.68 | 1.00 |
| 7.1 – 174.0 | 1.01 | 2.26 | 1.00 |
| 2.8 – <b>227.8</b> | 1.04 | 2.48 | 0.99 |

#### EliA Celikev IgG on Phadia 2500/5000

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The reportable range (Limit of Detection, upper limit) for EliA Celikey IgG is from 0.6 to 227 EliA U/mL. The measuring range (Limit of Quantitation, upper limit) is from 1.7 to 227 EliA U/mL.

| Dilution range<br>(EliA U/mL) | Slope | Intercept | R2 |
|-------------------------------|-------|-----------|------|
| 0.4 – 28.5 | 1.00 | -0.04 | 1.00 |
| 1.3 – 137.8 | 0.99 | -1.69 | 1.00 |
| 0.6 – 26.9 | 1.00 | -0.30 | 1.00 |
| 2.1 – 162.0 | 0.99 | 0.79 | 1.00 |

#### EliA Gliadin®º IgA on Phadia 2500/5000

The reportable range (Limit of Detection, upper limit) for EliA Gliadin® IgA is from 0.2 to 142 EliA U/mL. The measuring range (Limit of Quantitation, upper limit) is from 0.4 to 142 EliA U/mL. Please note that concentration values between LoD and LoQ may show a higher uncertainty.

#### EliA Gliadin®º IgG on Phadia 2500/5000

| Dilution range<br>(EliA U/mL) | Slope | Intercept | R2 |
|-------------------------------|-------|-----------|------|
| 0.6 - 26.3 | 0.99 | -0.32 | 1.00 |
| 0.8 – 54.4 | 1.00 | -0.50 | 1.00 |
| 7.0 - 334.9 | 0.98 | -5.65 | 0.99 |
| 7.3 – 288.3 | 1.00 | 1.02 | 1.00 |

The reportable range (Limit of Detection, upper limit) for EliA Gliadin® IgG is from 0.6 to 302 EliA U/mL. The measuring range (Limit of Quantitation, upper limit) is from 1.4 to 302 EliA U/mL.

Statements included in the package inserts: "Please note that concentration values between LoD and LoQ may show a higher uncertainty." "Please note that due to differing binding characteristics of the antibodies in patient samples, not all sera can be diluted linearly within the measuring range."

- Traceability, Stability, Expected values (controls, calibrators, or methods): C. The EliA IgG method was previously reviewed in K061165. The EliA IgA method was previously reviewed in K062787.
- d. Detection limit:

The limit of blank (LoB), limit of detection (LoD), and limit of quantitation studies were performed on the Phadia 2500/5000 instrument. One blank sample and three low level samples were measured in thirty-three and eleven replicates, respectively, in each of two runs.

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#### EliA Celikey IgG:

The LoD for EliA Celikey IgG is 0.6 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A2 and with proportions of false positives (a) less than 5% and false negatives (β) less than 5%; based on 132 determinations with 66 blank and 66 low level replicates; and LoB of 0.0 EliA U/mL. Since none of those low level samples met the target level for the CV% of 20% a requirement for the LoQ estimation - 3 additional low level samples were measured.

All low level samples were included in the LoD estimation (6 low level samples and a total of 132 determinations).

The LoQ for EliA Celikey IgG is 1.7 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A2, based on 66 determinations of 3 low level samples that are different from the ones for the determination of the LoD, and a target uncertainty goal of 20%.

The results are summarized in the table below:

| EliA Celikey IgG (EliA U/mL) | LoB | LoD | LoQ |
|------------------------------|-----|-----|-----|
| Phadia 2500/5000 | 0.0 | 0.6 | 1.7 |

#### EliA Gliadin®º IqA:

The LoD for EliA Gliadin®° IgA is 0.2 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A2 and with proportions of false positives (α) less than 5% and false negatives (β) less than 5%; based on 132 determinations with 66 blank and 66 low level replicates; and LoB of 0.0 ELiA U/mL.

The LoQ for Gliadin®º IgA is 0.4 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A2, based on 66 determinations, and a target uncertainty goal of 20%.

The results are summarized in the table below:

| EliA GliadinDP IgA (EliA U/mL) | LoB | LoD | LoQ |
|--------------------------------|-----|-----|-----|
| Phadia 2500/5000 | 0.0 | 0.2 | 0.4 |

#### EliA Gliadin®º IgG:

The LoD for EliA Gliadinºº IgG is 0.6 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A2 and with proportions of false positives (α) less than 5% and false negatives (β) less than 5%; based on 132 determinations with 66 blank and 66 low level replicates; and LoB of 0.3 EliA U/mL.

The LoQ for EliA Gliadinºº IgG is 1.4 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A2, based on 66 determinations, and a target uncertainty goal of 20%.

The results are summarized in the table below:

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| EliA GliadinDP IgG (EliA U/mL) | LoB | LoD | LoQ |
|--------------------------------|-----|-----|-----|
| Phadia 2500/5000 | 0.3 | 0.6 | 1.4 |

#### e. Analytical specificity:

Interference: Previously reviewed in K062583 and K093459. Carry-over: Phadia 2500/5000 instruments use disposable tips for pipetting samples and a separate pipette for the conjugate, therefore carry-over from samples to conjugate is impossible.

#### f. Assay cut-off:

The ranges (negative, equivocal, positive) recommended for the evaluation of the test results were derived from the clinical studies (s. K062583 and K093459).

#### EliA Celikey IqG Well

| < 7 EliA U/mL | Negative |
|------------------|-----------|
| 7 – 10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |

#### EliA Gliadin®º IgA Well

| < 7 EliA U/mL | Negative |
|------------------|-----------|
| 7 – 10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |

#### EliA Gliadin®º IgG Well

| < 7 EliA U/mL | Negative |
|------------------|-----------|
| 7 – 10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |

#### 2. Comparison studies:

- Method comparison with predicate device (Instrument comparison): a. See 2c Instrument Comparison below
- b. Matrix comparison:

Previously reviewed under K062583 and K093459.

#### Instrument comparison C.

In the Method Comparison studies for the three EliA tests included in this submission, more than 100 samples (≥20% of the samples within ±25% of the medical decision point) were run in single replicates on one Phadia 250 and three Phadia 2500/5000 instruments. The acceptance criteria for the method comparison (the slope for the regression lines should be 0.9 - 1.1 for single replicate to single replicate and intercept close to 0) were met for EliA Celikey IgG, EliA Gliadin®º IgA and EliA Gliadin®º IgG.

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## EliA Celikey lgG:

| Instrument | Intercept | 95% Cl | Slope | 95% Cl |
|---------------|-----------|----------------|-------|--------------|
| PH2500/5000 A | 0.49 | 0.26 to 0.73 | 0.93 | 0.91 to 0.94 |
| PH2500/5000 B | 0.14 | -0.19 to 0.46 | 0.96 | 0.93 to 1.00 |
| PH2500/5000 C | -0.28 | -0.55 to -0.00 | 0.98 | 0.97 to 1.00 |

# equivocal results considered positive

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|---------------|---------------|---------------|
| PPA | 97.4% | 97.4% | 96.1% |
| 95% CI | 90.9% - 99.7% | 90.9% - 99.7% | 89.0% - 99.2% |
| NPA | 82.6% | 87.0% | 91.3% |
| 95% CI | 61.2% - 95.0% | 66.4% - 97.2% | 72.0% - 98.9% |
| TPA | 94.0% | 95.0% | 95.0% |
| 95% CI | 87.4% - 97.8% | 88.7% - 98.4% | 88.7% - 98.4% |

#### equivocal results considered negative

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 95.5% | 97.0% | 94.0% |
| 95% CI | 87.5% - 99.1% | 89.6% - 99.6% | 85.4% - 98.3% |
| NPA | 100.0% | 100.0% | 100.0% |
| 95% CI | 89.4% - 100.0% | 89.4% - 100.0% | 89.4% - 100.0% |
| TPA | 97.0% | 98.0% | 96.0% |
| 95% CI | 91.5% - 99.4% | 93.0% - 99.8% | 90.1% - 98.9% |

{16}------------------------------------------------

# EliA Gliadin®° IgA:

| Instrument | Intercept | 95% Cl | Slope | 95% Cl |
|---------------|-----------|----------------|-------|--------------|
| PH2500/5000 A | 0.60 | 0.40 to 0.93 | 0.95 | 0.92 to 0.98 |
| PH2500/5000 B | 0.55 | 0.28 to 0.83 | 0.97 | 0.95 to 1.00 |
| PH2500/5000 C | -0.24 | -0.71 to -0.06 | 1.09 | 1.06 to 1.11 |

#### equivocal results considered positive

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 100.0% | 100.0% | 100.0% |
| 95% CI | 95.7% - 100.0% | 95.7% - 100.0% | 95.7% - 100.0% |
| NPA | 90.5% | 85.7% | 90.5% |
| 95% CI | 69.6% - 98.8% | 63.7% - 97.0% | 69.6% - 98.8% |
| TPA | 98.1% | 97.1% | 98.1% |
| 95% CI | 93.3% - 99.8% | 91.9% - 99.4% | 93.3% - 99.8% |

# equivocal results considered negative

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 98.7% | 100.0% | 98.7% |
| 95% CI | 93.2% - 100.0% | 95.4% - 100.0% | 93.2% - 100.0% |
| NPA | 96.2% | 96.2% | 96.2% |
| 95% CI | 80.4% - 99.9% | 80.4% - 99.9% | 80.4% - 99.9% |
| TPA | 98.1% | 99.0% | 98.1% |
| 95% CI | 93.3% - 99.8% | 94.8% - 100.0% | 93.3% - 99.8% |

# EliA Gliadin® IgG:

| Instrument | Intercept | 95% Cl | Slope | 95% Cl |
|---------------|-----------|---------------|-------|--------------|
| PH2500/5000 A | -0.14 | -0.41 to 0.13 | 1.06 | 1.04 to 1.09 |
| PH2500/5000 B | 0.43 | 0.13 to 1.16 | 0.99 | 0.94 to 1.02 |
| PH2500/5000 C | -0.06 | -0.20 to 0.20 | 1.08 | 1.07 to 1.10 |

{17}------------------------------------------------

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 100.0% | 100.0% | 100.0% |
| 95% CI | 95.8% - 100.0% | 95.8% - 100.0% | 95.8% - 100.0% |
| NPA | 89.5% | 100.0% | 94.7% |
| 95% CI | 66.9% - 98.7% | 82.4% - 100.0% | 74.0% - 99.9% |
| TPA | 98.1% | 100.0% | 99.0% |
| 95% CI | 93.2% - 99.8% | 96.5% - 100.0% | 94.8% - 100.0% |

equivocal results considered positive

equivocal results considered negative

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 98.8% | 100.0% | 100.0% |
| 95% CI | 93.2% - 100.0% | 95.5% - 100.0% | 95.5% - 100.0% |
| NPA | 100.0% | 95.8% | 91.7% |
| 95% CI | 85.8% - 100.0% | 78.9% - 99.9% | 73.0% - 99.0% |
| TPA | 99.0% | 99.0% | 98.1% |
| 95% CI | 94.8% - 100.0% | 94.8% - 100.0% | 93.2% - 99.8% |

- 3. Clinical studies:
- a. Clinical sensitivity: Not applicable.
- Clinical specificity: b. Not applicable.
- c. Other clinical supportive data (when a. and b. are not applicable): Clinical performance values were reviewed in K062583 and K093459.
- 4. Clinical cut-off: Same as assay cut-off.
- Expected values/Reference range: 5.

The frequency distribution for anti-tTG IgG and anti-gliadin IgA and IqG antibodies was investigated in a group of apparently healthy subjects equally distributed by age and gender, using sera from a Caucasian population obtained from a blood bank.

The results are given in the table below:

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| Test | n = | Median<br>(EliA U/mL) | 95th<br>percentile | 99th<br>percentile |
|-------------------------------------------|-----|-----------------------|--------------------|--------------------|
| EliA Celikey IgG on<br>Phadia 2500/5000 | 400 | 2.3 | 3.3 | 3.9 |
| EliA GliadinDP IgA on<br>Phadia 2500/5000 | 400 | 1.4 | 3.8 | 7.1 |
| EliA GliadinDP IgG on<br>Phadia 2500/5000 | 400 | 0.9 | 2.7 | 13.3 |

#### N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

#### O. Conclusion:

All available data support that both instrument platforms, Phadia 250 and Phadia 2500/5000 perform substantially equivalent when using the EliA Celikey IgG, EliA Gliadin®º IgA and EliA Gliadinºº IgG immunoassays.

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.