EliA CENP Immunoassay, EliA U1RNP Immunoassay, EliA RNP70 Immunoassay

K182353 · Phadia AB · LJM · Nov 27, 2018 · Immunology

Device Facts

• Record ID: K182353
• Device Name: EliA CENP Immunoassay, EliA U1RNP Immunoassay, EliA RNP70 Immunoassay
• Applicant: Phadia AB
• Product Code: LJM · Immunology
• Decision Date: Nov 27, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5100
• Device Class: Class 2

Intended Use

EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA CENP uses the EliA IgG method on the instrument Phadia 2500/5000. EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument Phadia 2500/5000. EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 uses the EliA IgG method on the instrument Phadia 2500/5000.

Device Story

EliA CENP, U1RNP, and RNP70 immunoassays are semi-quantitative in vitro diagnostic tests for IgG antibodies in human serum/plasma. The device uses polystyrene wells coated with specific recombinant antigens (CENP, U1RNP, or RNP70). Patient samples are incubated in these wells; specific antibodies bind to the antigen. After washing, enzyme-labeled anti-IgG conjugate is added, followed by a development solution. The Phadia 2500/5000 instrument measures the resulting fluorescence, which is proportional to the antibody concentration. The system is operated by laboratory personnel in a clinical setting. Results are compared to calibrators to determine antibody levels. The output aids clinicians in diagnosing connective tissue diseases. The device is not for point-of-care use.

Clinical Evidence

Bench testing only. Precision/reproducibility studies (21 runs, 84 replicates per sample) showed total imprecision (%CV) ranging from 6.0% to 24.4%. Linearity was confirmed with R2 values of 0.99-1.00. Method comparison studies (n > 100 samples) between Phadia 250 and Phadia 2500/5000 met acceptance criteria for slope (0.9-1.1) and intercept. LoD/LoQ were established per CLSI EP17-A. No new clinical diagnostic sensitivity/specificity studies were required as these were previously reviewed for the predicate assays.

Technological Characteristics

Solid-phase fluoroimmunoassay; recombinant protein-coated wells; β-Galactosidase labeled anti-IgG conjugate; 4-Methylumbelliferyl-β-D-galactoside development solution. Automated platform (Phadia 2500/5000) with disposable pipette tips. Incubation at 37°C. Connectivity via IDM software. Sterilization not applicable (reagents).

Indications for Use

Indicated for patients suspected of having scleroderma (CREST Syndrome), mixed connective tissue disease (MCTD), or systemic lupus erythematosus (SLE). Used as an aid in clinical diagnosis in conjunction with other laboratory and clinical findings. Not for point-of-care use.

Regulatory Classification

Identification

An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).

Predicate Devices

• EliA CENP on Phadia 250 (K082759)
• EliA U1RNP on Phadia 250 (K082759)
• EliA RNP70 on Phadia 250 (K083117)

Submission Summary (Full Text)

{0}------------------------------------------------

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the FDA logo is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

November 27, 2018

Phadia AB Sheryl Skinner Associate Director Phadia US Inc. 4169 Commercial Avenue Portage, Michigan 49002

Re: K182353

Trade/Device Name: EliA CENP Immunoassay EliA U1RNP Immunoassay EliA RNP70 Immunoassay Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear antibody immunological test system Regulatory Class: Class II Product Code: LJM, LKO Dated: August 28, 2018 Received: August 29, 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.

{1}------------------------------------------------

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,

# Leonthena R. Carrington -S

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) K182353

#### Device Name

EliA(TM) CENP Immunoassay, EliA(TM) U1RNP Immunoassay, EliA(TM) RNP70 Immunoassay

#### Indications for Use (Describe)

EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA IgG method on the instrument Phadia 2500/5000.

EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument Phadia 2500/5000.

EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 uses the EliA IgG method on the instrument Phadia 2500/5000.

| Type of Use (Select one or both, as applicable) | |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------|
| <span style="font-family: Arial, sans-serif;"> <span style="font-size: 10pt;"> <span style="color: black;"> <span style="font-weight: normal;"> <span style="font-style: normal;"> <span style="text-decoration: none;"> <span style="vertical-align: baseline;"> <span style="white-space: nowrap;"> <span style="word-spacing: 0px;"> <span style="letter-spacing: normal;"> <span style="text-transform: none;"> <span style="unicode-bidi: embed;"> </span> </span> </span> </span> </span> </span> </span> </span> </span> </span> </span> </span> Prescription Use (Part 21 CFR 801 Subpart D) | |
| | Over-The-Counter Use (21 CFR 801 Subpart C) |

#### CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

#### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.*

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

> Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

{3}------------------------------------------------

# 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:

K182353

#### B. Purpose for Submission:

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

#### C. Measurands:

CENP, U1RNP and RNP70 anti-nuclear antibodies

#### 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: Martin Mann Regulatory Affairs Manager Phadia US Inc. 4169 Commercial Avenue Portage, Mi 49002, USA +1 (-269-492) -1957 (Phone) +1 (-269-492) -7541 (Fax) [e-mail]

Date of Summary Preparation:

November 5, 2018

#### F. Proprietary and Established Names:

EliA CENP Immunoassay EliA U1RNP Immunoassay EliA RNP70 Immunoassay

{4}------------------------------------------------

#### G. Regulatory Information:

- 1. Regulation section: 21 CFR§ 866.5100, Antinuclear Antibody Immunological Test System
- 2. Classification: Class II

#### Product code: ဒေ

LJM Antinuclear antibody (enzyme-labeled), antigen, controls LKO Anti-RNP antibody, antigen and control

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

1. Intended use(s):

EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA CENP uses the EliA IgG method on the instrument Phadia 2500/5000.

EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument Phadia 2500/5000.

EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 uses the EliA IgG method on the instrument Phadia 2500/5000.

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

Special conditions for use statement(s): 3. For prescription use only

Special instrument requirements: ব

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

{5}------------------------------------------------

#### -Device Description:

The method-specific reagents are identical with K082759 (EliA CENP and EliA U1RNP) and K083117 (EliA RNP70), but are filled in containers specific for the Phadia 2500/5000 instrument. Each device consists of: Test Wells:

- EliA CENP Wells are coated with human recombinant centromere protein B 2 carriers (12 wells each), ready to use;
- -EliA U1RNP Wells are coated with human recombinant RNP (RNP70, A. C) proteins- 4 carriers (12 wells each), ready to use;
- -EliA RNP70 Wells are coated with human recombinant RNP (70 kDa) protein-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:

- 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.

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 CENP, EliA U1RNP and EliA RNP70 tests.

#### J. Substantial Equivalence Information:

1. Predicate device name(s) and 510(k) number(s): EliA CENP on Phadia 250 instrument, K082759 EliA U1RNP on Phadia 250 instrument, K082759 EliA RNP70 on Phadia 250 instrument, K083117

{6}------------------------------------------------

#### 2. Comparison with predicate device:

#### EliA CENP, EliA U1RNP and EliA RNP70 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 CENP | EliA CENP is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to CENP in human serum<br>and plasma (heparin, EDTA,<br>citrate) as an aid in the clinical<br>diagnosis of scleroderma (CREST<br>Syndrome) in conjunction with<br>other laboratory and clinical<br>findings. EliA CENP uses the EliA<br>IgG method on the instrument<br>ImmunoCAP 250. | EliA CENP is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to CENP in human<br>serum and plasma (Li-heparin,<br>EDTA) as an aid in the clinical<br>diagnosis of scleroderma<br>(CREST Syndrome) in<br>conjunction with other laboratory<br>and clinical findings. EliA CENP<br>uses the EliA IgG method on the<br>instrument Phadia 2500/5000. |
| Intended Use<br>EliA U1RNP | EliA U1RNP is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to U1RNP in human<br>serum and plasma (heparin,<br>EDTA, citrate) as an aid in the<br>clinical diagnosis of mixed<br>connective tissue disease (MCTD)<br>and systemic lupus<br>erythematosus (SLE) in<br>conjunction with other laboratory<br>and clinical findings. EliA U1RNP<br>uses the EliA IgG method on the<br>instrument ImmunoCAP 250. | EliA U1RNP is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to U1RNP in human<br>serum and plasma (Li-heparin,<br>EDTA) as an aid in the clinical<br>diagnosis of mixed connective<br>tissue disease (MCTD) and<br>systemic lupus erythematosus<br>(SLE) in conjunction with other<br>laboratory and clinical findings.<br>EliA U1RNP uses the EliA IgG<br>method on the instrument<br>Phadia 2500/5000. |
| Intended Use<br>EliA RNP70 | EliA RNP70 is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to RNP70 in human<br>serum and plasma (heparin,<br>EDTA) as an aid in the clinical<br>diagnosis of mixed connective<br>tissue disease (MCTD) and<br>systemic lupus erythematosus<br>(SLE) in conjunction with other<br>laboratory and clinical findings.<br>EliA RNP70 is to be used together<br>with the EliA IgG method on the<br>instrument ImmunoCAP 250 | EliA RNP70 is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to RNP70 in human<br>serum and plasma (Li-heparin,<br>EDTA) as an aid in the clinical<br>diagnosis of mixed connective<br>tissue disease (MCTD) and<br>systemic lupus erythematosus<br>(SLE) in conjunction with other<br>laboratory and clinical findings.<br>EliA RNP70 uses the EliA IgG<br>method on the instrument<br>Phadia 2500/5000 |
| Analytical | Same | Same |
| technology: | | |
| Immuno- | | |
| fluorescence | | |
| measurement | | |
| Assay process | Same | Same |
| Common,<br>dedicated Phadia | Same | Same<br>Introduction of new article |
| reagents | | numbers for Development<br>Solution, Stop Solution and |
| | | Washing Solution is only due to<br>larger filling volumes which are |
| | | required for the bigger<br>instruments Phadia 2500/5000 |
| Result calculation | Same | Same |
| software; Phadia | | |
| Information Data | | |
| Manager (IDM) | | |
| 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 | 90 µL | 90 µL |
| Solution Volume | | |
| Stop Solution<br>Volume | 200 µL | 200 µL |
| Assay set-up | Random access | Random access |
| Reagent<br>packaging size | Various/Common | Various/Common<br>Introduction of new article |
| | | number for EliA Sample Diluent |
| | | (83-1071-01) is only due to<br>larger filling volume. |
| Onboard storage | Yes | Yes |
| of reagents | | |
| | | |
| 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 | Human serum or plasma<br>(heparin, EDTA, citrate) | Human serum or plasma (Li-<br>heparin, EDTA); i.e. citrate<br>plasma is omitted for all tests |
| 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. |

{7}------------------------------------------------

{8}------------------------------------------------

## EliA CENP, EliA U1RNP and EliA RNP70 Immunoassays on Phadia 250 and Phadia 2500/5000 instruments – Differences to predicate device

{9}------------------------------------------------

# 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 centromere protein B, human recombinant RNP (RNP70, A, C) or with human recombinant RNP (70 kDa) protein. If present in the patient's specimen, antibodies to these proteins bind to the specific antigen. After washing away non-bound antibodies, enzyme-labeled antibodies against human IgG antibodies (EliA IgG Conjugate) are added to form an antibodyconjugate 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 assay directly measures the amount of antibody of interest bound to the antigen coating the EliA well, therefore the higher the value of fluorescent signal detected by the instrument, the higher the amount of antibody bound and detected in the sample tested. 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/day 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.

{10}------------------------------------------------

The results are summarized in the table below:

| Mean | Within-Run | | Between-<br>Run | | Between-<br>Instrument | | Total<br>Imprecision | |
|----------------|------------|-----|-----------------|-----|------------------------|------|----------------------|------|
| (EliA<br>U/mL) | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 1.1 | 0.1 | 9.9 | 0.1 | 6.1 | 0.2 | 15.8 | 0.2 | 19.6 |
| 7.1 | 0.4 | 5.8 | 0.3 | 4.9 | 0.3 | 3.9 | 0.6 | 8.6 |
| 11.4 | 0.4 | 3.5 | 0.4 | 3.9 | 0.5 | 4.2 | 0.8 | 6.7 |
| 129.8 | 4.3 | 3.3 | 5.1 | 3.9 | 3.9 | 3.0 | 7.7 | 6.0 |
| 229.7 | 9.8 | 4.3 | 15.7 | 6.8 | 10.1 | 4.4 | 21.1 | 9.2 |

#### EliA CENP on Phadia 2500/5000

#### EliA U1RNP on Phadia 2500/5000

| Mean<br>(EliA<br>U/mL) | Within-Run | | Between-Run | | Between-Instrument | | Total<br>Imprecision | |
|------------------------|------------|------|-------------|------|--------------------|------|----------------------|------|
| | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 2.2 | 0.4 | 17.9 | 0.3 | 12.5 | 0.2 | 11.0 | 0.5 | 24.4 |
| 5.7 | 0.3 | 5.4 | 0.2 | 2.8 | 0.1 | 1.8 | 0.4 | 6.4 |
| 10.4 | 0.5 | 4.7 | 0.4 | 4.2 | 0.6 | 5.6 | 0.9 | 8.5 |
| 135.4 | 5.2 | 3.8 | 7.2 | 5.3 | 6.5 | 4.8 | 11.0 | 8.1 |
| 231.7 | 9.1 | 3.9 | 9.0 | 3.9 | 10.7 | 4.6 | 16.7 | 7.2 |

#### EliA RNP70 on Phadia 2500/5000

| Mean<br>(EliA<br>U/mL) | Within-Run | | Between-<br>Run | | Between-<br>Instrument | | Total<br>Imprecision | |
|------------------------|------------|-----|-----------------|-----|------------------------|------|----------------------|------|
| | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 1.4 | 0.1 | 5.4 | 0.1 | 4.4 | 0.2 | 17.0 | 0.3 | 18.4 |
| 7.9 | 0.2 | 2.9 | 0.3 | 4.4 | 0.5 | 6.2 | 0.6 | 8.1 |
| 8.9 | 0.5 | 5.2 | 0.5 | 5.5 | 0.5 | 5.2 | 0.8 | 9.2 |
| 93.1 | 5.1 | 5.5 | 5.5 | 5.9 | 2.9 | 3.1 | 8.0 | 8.6 |
| 190.2 | 9.4 | 5.0 | 10.3 | 5.4 | 10.6 | 5.6 | 17.5 | 9.2 |

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

Four patient serum samples were diluted in sample diluent and tested with one batch of EliA CENP, EliA U1RNP and EliA RNP70 Immunoassays and one set of system reagents on Phadia 2500/5000. The results of the regression analysis are summarized below:

{11}------------------------------------------------

| Dilution range<br>(EliA U/mL) | Slope | 95% CI | Intercept | 95% CI | R2 |
|-------------------------------|-------|-------------|-----------|---------------|------|
| 0.5 – 24.3 | 1.00 | 0.99 – 1.01 | 0.17 | 0.02 – 0.31 | 1.00 |
| 2.1 – 180.3 | 1.01 | 0.99 – 1.02 | 0.10 | -0.83 – 1.03 | 1.00 |
| 3.8 – 305.0 | 1.04 | 0.99 – 1.10 | 2.70 | -4.21 – 9.62 | 0.99 |
| 0.4 – 18.8 | 0.99 | 0.97 – 1.02 | -0.22 | -0.44 – -0.00 | 1.00 |

#### EliA CENP on Phadia 2500/5000

The reportable range (Limit of Detection, upper limit) for EliA CENP is from 0.4 to 240 EliA U/mL. The measuring range (Limit of Quantitation, upper limit) is from 0.7 to 240 EliA U/mL.

#### EliA U1RNP on Phadia 2500/5000

| Dilution range<br>(EliA U/mL) | Slope | 95% CI | Intercept | 95% CI | R2 |
|-------------------------------|-------|-------------|-----------|--------------|------|
| 0.5 – 18.0 | 1.01 | 0.99 - 1.02 | 0.08 | -0.04 - 0.20 | 1.00 |
| 0.6 – 51.3 | 1.01 | 0.99 – 1.02 | 0.04 | -0.20 - 0.27 | 1.00 |
| 0.6 – 15.8 | 1.00 | 0.96 - 1.04 | 0.28 | 0.01 - 0.56 | 1.00 |
| 4.7 – 284.7 | 1.04 | 0.98 - 1.10 | 1.86 | -4.75 - 8.47 | 0.99 |

The reportable range (Limit of Detection, upper limit) for EliA U1RNP is from 0.5 to 240 EliA U/mL. The measuring range (Limit of Quantitation, upper limit) is from 1.1 to 240 EliA U/mL.

| Dilution range<br>(EliA U/mL) | Slope | 95% CI | Intercept | 95% CI | R2 |
|-------------------------------|-------|-------------|-----------|--------------|------|
| 0.6 - 62.5 | 1.02 | 1.00 – 1.04 | 0.07 | -0.47 - 0.49 | 1.00 |
| 0.4 - 38.7 | 1.01 | 0.98 – 1.04 | -0.42 | -0.87 - 0.02 | 1.00 |
| 6.9 - 251.8 | 1.00 | 0.97 – 1.03 | 3.07 | 0.05 - 6.09 | 1.00 |
| 3.7 - 294.6 | 1.01 | 0.98 – 1.03 | 3.38 | 0.39 - 6.38 | 1.00 |

#### EliA RNP70 on Phadia 2500/5000

The reportable range (Limit of Detection, upper limit) for EliA RNP70 is from 0.3 to 240 EliA U/mL. The measuring range (Limit of Quantitation, upper limit) is from 0.9 to 240 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.

{12}------------------------------------------------

#### d. Detection limit:

The limit of blank (LoB) and limit of detection (LoD) 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.

#### EliA CENP:

The LoD for EliA CENP is 0.4 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A 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 0.2 EliA U/mL.

The LoQ for EliA CENP is 0.7 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A, based on 66 determinations, and a target uncertainty goal of 20%.

The results are summarized in the table below:

| EliA CENP (EliA U/mL) | LoB | LoD | LoQ |
|-----------------------|-----|-----|-----|
| Phadia 2500/5000 | 0.2 | 0.4 | 0.7 |

#### EIiA U1RNP:

The LoD for EliA U1RNP is 0.5 EliA U/mL, determined consistent with the quidelines in CLSI document EP17-A 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 0.2 EliA U/mL.

The LoQ for EliA U1RNP is 1.1 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A, based on 66 determinations, and a target uncertainty goal of 20%.

The results are summarized in the table below:

| EliA U1RNP (EliA U/mL) | LoB | LoD | LoQ |
|------------------------|-----|-----|-----|
| Phadia 2500/5000 | 0.2 | 0.5 | 1.1 |

#### EliA RNP70:

The LoD for EliA RNP70 is 0.3 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A 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 0.2 EliA U/mL.

The LoQ for EliA RNP70 is 0.9 EliA U/mL, determined consistent with the guidelines in CLSI document EP17-A, based on 66 determinations, and a target uncertainty goal of 20%.

{13}------------------------------------------------

The results are summarized in the table below:

| EliA RNP70 (EliA U/mL) | LoB | LoD | LoQ |
|------------------------|-----|-----|-----|
| Phadia 2500/5000 | 0.2 | 0.3 | 0.9 |

- e. Analytical specificity:
Interference: Previously reviewed in K082759 and K083117.

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. K082759 and K083117).

#### EliA CENP Well

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

#### EliA U1RNP Well

| < 5 EliA U/mL | Negative |
|------------------|-----------|
| 5 - 10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |

#### EliA RNP70 Well

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

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

Previously reviewed under K082759 and K083117.

- 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 CENP, EliA U1RNP and EliA RNP70.

{14}------------------------------------------------

### EliA CENP:

| Instrument | Intercept | 95% Cl | Slope | 95% Cl |
|---------------|-----------|----------------|-------|--------------|
| PH2500/5000 A | -0.30 | -0.55 to -0.07 | 1.02 | 1.00 to 1.03 |
| PH2500/5000 B | 0.44 | 0.07 to 0.78 | 0.98 | 0.95 to 1.00 |
| PH2500/5000 C | -0.40 | -0.91 to -0.09 | 1.04 | 1.03 to 1.07 |

#### 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% | 95.5% - 100.0% |
| NPA | 94.1% | 88.2% | 88.2% |
| 95% CI | 71.3% - 99.9% | 63.6% - 98.5% | 63.6% - 98.5% |
| TPA | 99.0% | 98.0% | 97.9% |
| 95% CI | 94.6% - 100.0% | 93.0% - 99.8% | 92.7% - 99.7% |

### equivocal results considered negative

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 98.7% | 98.7% | 98.6% |
| 95% CI | 92.8% - 100.0% | 92.8% - 100.0% | 92.5% - 100.0% |
| NPA | 100.0% | 92.0% | 96.0% |
| 95% CI | 86.3% - 100.0% | 74.0% - 99.0% | 79.6% - 99.9% |
| TPA | 99.0% | 97.0% | 97.9% |
| 95% CI | 94.6% - 100.0% | 91.5% - 99.4% | 92.7% - 99.7% |

{15}------------------------------------------------

# EliA U1RNP:

| Instrument | Intercept | 95% Cl | Slope | 95% Cl |
|---------------|-----------|---------------|-------|--------------|
| PH2500/5000 A | 0.68 | 0.34 to 1.02 | 0.92 | 0.90 to 0.95 |
| PH2500/5000 B | 0.03 | -0.36 to 0.65 | 1.02 | 0.98 to 1.05 |
| PH2500/5000 C | 0.98 | 0.43 to 1.26 | 0.92 | 0.90 to 0.94 |

#### equivocal results considered positive

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 100.0% | 98.9% | 96.8% |
| 95% CI | 96.2% - 100.0% | 94.3% - 100.0% | 91.0% - 99.3% |
| NPA | 100.0% | 94.1% | 100.0% |
| 95% CI | 80.5% - 100.0% | 71.3% - 99.9% | 80.5% - 100.0% |
| TPA | 100.0% | 98.2% | 97.3% |
| 95% CI | 96.8% - 100.0% | 93.7% - 99.8% | 92.4%- 99.4% |

### equivocal results considered negative

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|---------------|----------------|
| PPA | 98.7% | 97.4% | 100.0% |
| 95% CI | 93.0% - 100.0% | 90.9% - 99.7% | 95.3% - 100.0% |
| NPA | 97.1% | 97.1% | 97.1% |
| 95% CI | 85.1% - 99.9% | 85.1% - 99.9% | 85.1% - 99.9% |
| TPA | 98.2% | 97.3% | 99.1% |
| 95% CI | 93.7% - 99.8% | 92.4% - 99.4% | 95.1% - 100.0% |

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

#### EliA RNP70:

| Instrument | Intercept | 95% Cl | Slope | 95% Cl |
|---------------|-----------|--------------|-------|--------------|
| PH2500/5000 A | 0.35 | 0.17 to 0.52 | 1.00 | 0.98 to 1.02 |
| PH2500/5000 B | 0.29 | 0.17 to 0.48 | 1.04 | 1.01 to 1.06 |
| PH2500/5000 C | 0.33 | 0.16 to 0.69 | 1.03 | 1.01 to 1.06 |

#### equivocal results considered positive

| 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 | 88.5% | 80.8% | 84.6% |
| 95% CI | 69.8% - 97.6% | 60.6% - 93.4% | 65.1% - 95.6% |
| TPA | 97.3% | 95.5% | 96.4% |
| 95% CI | 92.3% - 99.4% | 89.8% - 98.5% | 91.1% - 99.0% |

#### equivocal results considered negative

| criteria | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
|----------|----------------|----------------|----------------|
| PPA | 98.6% | 100.0% | 98.6% |
| 95% CI | 92.6% - 100.0% | 95.1% - 100.0% | 92.7% - 100.0% |
| NPA | 97.4% | 92.1% | 92.1% |
| 95% CI | 86.2% - 99.9% | 78.6% - 98.3% | 78.6% - 98.3% |
| TPA | 98.2% | 97.3% | 96.4% |
| 95% CI | 93.6% - 99.8% | 92.3% - 99.4% | 91.1% - 99.0% |

#### 3. Clinical studies:

- Clinical sensitivity: a. Not applicable.
- b. Clinical specificity: Not applicable.
- c. Other clinical supportive data (when a. and b. are not applicable): Clinical performance values were reviewed in K082759 and K083117.
- 4. Clinical cut-off:

Same as assay cut-off.

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

#### 5. Expected values/Reference range:

The frequency distribution for CENP, U1RNP and RNP70 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.

| Test | n = | Median<br>(EliA U/mL) | 95th<br>percentile | 99th<br>percentile |
|-----------------------------------|-----|-----------------------|--------------------|--------------------|
| EliA CENP on Phadia<br>2500/5000 | 400 | 0.7 | 1.0 | 1.3 |
| EliA U1RNP on<br>Phadia 2500/5000 | 400 | 1.5 | 2.2 | 3.0 |
| EliA RNP70 on<br>Phadia 2500/5000 | 400 | 1.4 | 2.2 | 2.5 |

The results are given in the table below:

#### 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 CENP, EliA U1RNP and EliA RNP70 immunoassays.

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