EliA Ro52, EliA Ro60

K210902 · Phadia AB · LKJ · Jul 27, 2022 · Immunology

Device Facts

• Record ID: K210902
• Device Name: EliA Ro52, EliA Ro60
• Applicant: Phadia AB
• Product Code: LKJ · Immunology
• Decision Date: Jul 27, 2022
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5100
• Device Class: Class 2

Intended Use

EliA Ro52 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Ro52 in human serum as an aid in the diagnosis of Sjögren's syndrome (SS), systemic lupus erythematosus (SLE), systemic sclerosis (SSc) and idiopathic inflammatory myopathies (IIM) in conjunction with other laboratory and clinical findings. EliA Ro52 uses the EliA IgG method. EliA Ro60 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Ro60 in human serum as an aid in the diagnosis of Sjögren's syndrome (SS) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA Ro60 uses the EliA IgG method.

Device Story

EliA Ro52 and EliA Ro60 are automated, solid-phase, fluoroenzyme immunoassays for detecting IgG autoantibodies against SS-A/Ro 52 kDa and 60 kDa proteins. The system uses polystyrene wells coated with human recombinant antigens. Patient serum is added; specific antibodies bind to the antigen. After washing, enzyme-labeled anti-human IgG conjugate is added, followed by a development solution. The instrument measures fluorescence, which is proportional to the amount of bound antibody. Results are compared to a calibration curve. The device is used in clinical laboratories on Phadia 250, 2500, and 5000 series instruments. Healthcare providers use these results alongside clinical findings to diagnose autoimmune conditions. The device benefits patients by providing standardized, automated, and reproducible antibody quantification to support clinical diagnosis.

Clinical Evidence

Clinical performance was evaluated using 755 (Ro52) and 713 (Ro60) serum samples from target disease groups (SLE, SS, IIM, SSc) and control groups (autoimmune/infectious diseases). For EliA Ro52, sensitivity ranged from 20.9% to 55.0% and specificity from 95.6% to 96.9% depending on the target disease. For EliA Ro60, sensitivity ranged from 48.3% to 71.7% and specificity from 98.4% to 98.6%. Analytical performance included precision (CVs <10%), linearity, and interference testing.

Technological Characteristics

Solid-phase fluoroenzymeimmunoassay (FEIA). Wells coated with human recombinant SS-A/Ro 52 kDa or 60 kDa protein. Detection via β-Galactosidase labeled anti-human IgG conjugate and 4-Methylumbelliferyl-β-D-galactoside substrate. Fluorescence measurement. Automated system (Phadia 250/2500/5000). Calibrated to WHO IRP 67/86. Analytical measuring range: 0.7–240 EliA U/mL (Ro52) and 0.5–240 EliA U/mL (Ro60).

Indications for Use

Indicated for the semi-quantitative measurement of IgG antibodies to Ro52 or Ro60 in human serum to aid in the diagnosis of Sjögren's syndrome (SS), systemic lupus erythematosus (SLE), systemic sclerosis (SSc), and idiopathic inflammatory myopathies (IIM) in conjunction with other clinical and laboratory findings. For prescription use only.

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

• Quanta Flash Ro52 (K141655)
• Quanta Flash Ro60 (K141328)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: a symbol on the left and the FDA name and title on the right. The symbol on the left is a stylized representation of a human figure, while the text on the right reads "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue letters.

July 27, 2022

Phadia AB % Jane Anthony Senior Manager, Quality Systems and Compliance Phadia US Inc. 4169 Commercial Avenue Portage, Michigan 49002

Re: K210902

Trade/Device Name: EliA Ro52 EliA Ro60 Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear Antibody Immunological Test System Regulatory Class: Class II Product Code: LKJ Dated: April 4, 2022 Received: April 6, 2022

Dear Jane Anthony:

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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542 of the Act); 21 CFR 1000-1050.

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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); 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-

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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Ying Mao, Ph.D. Branch Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known)

K210902

Device Name

EliA Ro52

#### Indications for Use (Describe)

EliA Ro52 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Ro52 in human serum as an aid in the diagnosis of Sjögren's syndrome (SS), systemic lupus erythematosus (SLE), systemic sclerosis (SSc) and idiopathic inflammatory myopathies (IIM) in conjunction with other laboratory and clinical findings. EliA Ro52 uses the EliA IgG method.

| Type of Use (Select one or both, as applicable) | |
|-------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------|
| <span style="text-decoration: underline;">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) | <span style="text-decoration: underline;">☐</span> Over-The-Counter Use (21 CFR 801 Subpart C) |

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

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# Indications for Use

510(k) Number (if known)

K210902

Device Name

EliA Ro60

#### Indications for Use (Describe)

EliA Ro60 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Ro60 in human serum as an aid in the diagnosis of Sjögren's syndrome (SS) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA Ro60 uses the EliA IgG method.

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

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

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

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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 [e-mail]

"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."

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# 510(K) DECISION SUMMARY

This 510(k) Summary is prepared in accordance with the requirements of 21 CFR Part 807.92.

### Premarket Notification 510(k) No: K210902

| Date of Summary Preparation: | July 25, 2022 |
|------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Manufacturer: | Phadia AB<br>Rapsgatan 7P<br>P.O. Box 6460<br>751 37 Uppsala, Sweden |
| Distributor: | Phadia US Inc.<br>4169 Commercial Avenue<br>Portage, MI 49002 |
| Company Contact Person: | Jane Anthony<br>Senior Manager, Quality Systems and Compliance<br>Phadia US Inc.<br>4169 Commercial Avenue, Portage, MI 49002<br>269-254-6833<br>[e-mail] |

### Proprietary and Established Device Name:

EliA Ro52 EliA Ro60

# Regulatory Information:

| Product Code: | LKJ |
|-----------------|---------------------------------------------------------------------|
| Classification: | Class II |
| Regulation: | 21 CFR 866.5100 – Antinuclear Antibody Immunological Test<br>System |
| Panel: | Immunology |

# Purpose of Submission:

New Device

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### Measurand:

IgG autoantibodies specific to SS-A/Ro proteins (52 kDa and 60 kDa)

# Type of Test:

Automated semi-quantitative solid phase fluoroenzymeimmunoassay.

# Intended Use:

EliA Ro52 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Ro52 in human serum as an aid in the diagnosis of Sjögren's syndrome (SS), systemic lupus erythematosus (SLE), systemic sclerosis (SSc) and idiopathic inflammatory myopathies (IIM) in conjunction with other laboratory and clinical findings. EliA Ro52 uses the EliA IgG method.

EliA Ro60 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Ro60 in human serum as an aid in the diagnosis of Sjögren's syndrome (SS) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA Ro60 uses the EliA IgG method.

# Indication(s) for Use:

Same as intended use

### Special Conditions for Use:

Rx - For Prescription Use Only

# Special Instrument Requirements:

For use on the Phadia 250 instrument and the Phadia 5000 instrument series (E-modules).

# Device Description:

The EliA Ro52 and EliA Ro60 Immunoassays are semi-quantitative solid-phase fluoroenzyme immunoassays, for the determination of autoantibodies against SS-A/Ro 52 kDa and 60 kDa proteins. The EliA Ro52 and EliA Ro60 test System is a fully integrated and automated system composed of assay-specific reagents, EliA method-specific reagents, and general reagents.

### Assay-Specific Reagents include:

- . EliA Ro52 Wells: coated with human recombinant SS-A/Ro (52 kDa) protein - 2 carriers (12 wells each), ready to use.
- EliA Ro60 Wells: coated with human recombinant SS-A/Ro (60 kDa) protein ■ - 4 carriers (12 wells each), ready to use.

Phadia AB, Rapsgatan 7P, P.O. Box 6460, 751 37 Uppsala, Sweden

Confidential Page 2 (26)

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- . EliA ANA Positive Control 250 or 2500/5000: Human serum in PBS containing IgG antibodies to dsDNA, RNP, Sm, Ro. La, Scl-70, CENP, and Jo-1 - 6 single use vials, 0.3 mL each, ready to use.
- . EliA ANA 3 Positive Control 250 or 2500/5000: Human monoclonal antibodies in PBS containing BSA, detergent and sodium azide (0.095% (w/v)) containing IgG antibodies to Ro52, Rib-P and RNA Pol III - 6 single use vials, 0.3 mL each, ready to use.
- . EliA IgG/IgM/IgA Negative Control 250 or 2500/5000: Human blood preparation from healthy donors in PBS containing BSA, detergent and 0.095% sodium azide - 6 singleuse vials, 0.3 mL each, ready to use.

# EliA Method-Specific Reagents include:

- I EliA Sample Diluent: PBS containing BSA. detergent and 0.095% sodium azide - 6 bottles, 48 mL each, ready to use; or 6 bottles, 400 mL each, ready to use.
- I EliA IgG Conjugate 50 or 200: ß-Galactosidase labeled anti-IgG (mouse monoclonal antibodies) in PBS containing BSA and 0.06% sodium azide - 6 wedge shaped bottles, 5 mL each, ready to use; or 6 wedge shaped bottles, 19 mL each, ready to use.
- 트 EliA IgG Calibrator Strips: Human IgG (0, 4, 10, 20, 100, 600 µg/L) in PBS containing BSA, detergent and 0.095% sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use.
- I EliA IgG Curve Control Strips: Human IgG (20 µg/L) in PBS containing BSA, detergent and 0.095% sodium azide - 5 strips, 6 single-use vials per strip, 0.3 mL each, ready to use.
- I EliA IgG Calibrator Well: coated with mouse monoclonal antibodies - 4 carriers (12 wells each), ready to use.

# General Reagents include:

- Development Solution: 0.01% 4-Methylumbelliferyl-B-D-galactoside, <0.0010% . preservative - 6 bottles (11 mL, 17 mL, or 112 mL each), sufficient for 6x >110, 6x >170, or 6x >1165 determinations.
- I Stop Solution: 4% Sodium Carbonate - 6 bottles (65 mL, 119 mL, or 2800 mL each), sufficient for 6x >292, 6x >560, or 6x >13100 determinations.
- I Washing Solution Additive: detergent, preservative <0.13% - 6x 17.2 mL, 2x 86mL, or 4x 850 mL.
- 트 Washing Solution Concentrate: phosphate buffer - 6x 80 mL, 2x 400 mL, or 1x 2800 mL.

### Instrument System

The EliA Ro52 and EliA Ro60 Immunoassays are run on the Phadia 250 instrument and the Phadia 2500 and 5000 instrument series. The instruments are automated platforms for EliA test procedures from sample and reagent handling to the processing of results.

# Substantial Equivalence

QUANTA Flash Ro52, Inova Diagnostics Inc. (K141655) QUANTA Flash Ro60, Inova Diagnostics Inc. (K141328)

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# Comparison with Predicate Device EliA Ro52:

| | EliA Ro52<br>(Proposed Device) | QUANTA Flash Ro52 |
|--------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | | |
| Similarities | | |
| Assay Intended Use | EliA Ro52 is intended for the in<br>vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to Ro52 in human<br>serum as an aid in the diagnosis<br>of Sjögren's syndrome (SS),<br>systemic lupus erythematosus<br>(SLE), systemic sclerosis (SSc)<br>and idiopathic inflammatory<br>myopathies (IIM) in conjunction<br>with other laboratory and<br>clinical findings. EliA Ro52<br>uses the EliA IgG method. | QUANTA Flash® Ro52 is a<br>chemiluminescent<br>immunoassay for the semi-<br>quantitative determination of<br>IgG anti-Ro52 autoantibodies<br>in human serum. The presence<br>of anti-Ro52 autoantibodies, in<br>conjunction with clinical<br>findings and other laboratory<br>tests, is an aid in the diagnosis<br>of Systemic Lupus<br>Erythematosus (SLE),<br>Sjögren's Syndrome, Systemic<br>Sclerosis, Idiopathic<br>Inflammatory Myopathies. |
| Classification | Class II | Class II |
| Regulation Number | 866.5100 | 866.5100 |
| Internal Controls | Positive and negative Control<br>provided with EliA ANA 3<br>Positive Control 250 /<br>2500/5000 and EliA<br>IgG/IgM/IgA Negative Control<br>250/2500/5000, respectively. | QUANTA Flash® Ro52 Controls<br>(Negative and Positive Control) |
| Assay technique | ELISA | ELISA |
| Type of test | Semi-quantitative | Semi-quantitative |
| Reaction<br>temperature | 37°C controlled | 37°C controlled |
| Differences | | |
| Antigen | Human recombinant SS-A/Ro<br>52 (kDa) protein | Purified recombinant Ro52<br>antigen |
| Product Code | LKJ | OBE |
| Sample Dilution<br>(taking a 1%<br>pipetting<br>imprecision into<br>consideration, this<br>sample dilution is<br>regarded as a | 1:100 | 1:23 |
| | EliA Ro52<br>(Proposed Device) | QUANTA Flash Ro52 |
| Reporting of results | EliA U/mL (arbitrary) | Chemiluminescent units (CU) |
| Instrumentation | EliA Ro52 uses the EliA IgG<br>method on the instruments<br>Phadia 250 and the E-Modules<br>of the Phadia 2500 and Phadia<br>5000 series. | Quanta Flash Ro52 is run on<br>the BIO-FLASH® instrument. |
| Detection antibody<br>(conjugate) | IgG conjugate: anti-human IgG<br>ß-Galactosidase (mouse<br>monoclonal antibodies) | Isoluminol conjugated anti-<br>human IgG |
| Signal | Fluorescence | Relative Light Units (RLU) |
| Calibration | 6-point total IgG Calibration<br>6 vials of human IgG at<br>concentrations of<br>0–4–10–20–100–600 µg/L | Lot specific Master Curve and<br>two Calibrators<br>(Sold separately) |
| Calibration curve | Option to store curve for up to<br>28 days and run curve controls<br>in each assay for calibration | These calibrators are designed<br>for 4 calibrations. The total<br>time the calibrator tubes can be<br>uncapped onboard the<br>instrument is 8 hours. If the<br>calibrators are left uncapped,<br>onboard, for any longer period<br>of time, they should be<br>discarded. Using the same<br>calibrator tubes for more than 8<br>hours can result in improper<br>calibration of the assay, which<br>in turn could give erroneous<br>results. |
| Interpretation of<br>results | Negative < 7 EliA U/mL<br>Equivocal 7-10 EliA U/mL<br>Positive > 10 EliA U/mL | Negative: < 20 CU Positive: > 20<br>CU |
| Substrate | Development Solution 0.01 %<br>4-Methylumbelliferyl-ß-D-<br>galactoside,<br><0.0010% preservative*<br>*Preservative: mixture of 5-<br>chloro-2-methyl-2H-isothiazol-<br>3-one [EC no. 247-500-7] and 2-<br>methyl-2H-isothiazol-3-one [EC<br>no. 220-239-6] (3:1) | "Trigger" reagents |

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# SCIENTIF

Traditional 510(k) for EliA Ro52 and EliA Ro60, Amendment to K210902 A.7 510(k) Decision Summary_V4

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# Comparison with Predicate Device EliA Ro60:

| | EliA Ro60<br>(Proposed Device) | QUANTA Flash Ro60 |
|-----------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Similarities | | |
| Assay Intended Use | EliA Ro60 is intended for the in vitro semi-quantitative<br>measurement of IgG antibodies<br>directed to Ro60 in human<br>serum as an aid in the diagnosis<br>of Sjögren's syndrome (SS) and<br>systemic lupus erythematosus<br>(SLE) in conjunction with other<br>laboratory and clinical findings.<br>EliA Ro60 uses the EliA IgG<br>method. | QUANTA Flash Ro60 is a<br>chemiluminescent<br>immunoassay for the semi-<br>quantitative determination of<br>IgG anti-Ro60 autoantibodies<br>in human serum. The presence<br>of anti-Ro60 autoantibodies, in<br>conjunction with clinical<br>findings and other laboratory<br>tests, is an aid in the diagnosis<br>of Systemic Lupus<br>Erythematosus and Sjögren's<br>Syndrome. |
| Classification | Class II | Class II |
| Regulation Number | 866.5100 | 866.5100 |
| Internal Controls | Positive and negative Control<br>provided with EliA ANA 3<br>Positive Control 250 /<br>2500/5000 and EliA<br>IgG/IgM/IgA Negative Control<br>250 / 2500/5000, resp. | QUANTA Flash® Ro60<br>Controls (Negative and Positive<br>Control) |
| Assay technique | ELISA | ELISA |
| Type of test | Semi-quantitative | Semi-quantitative |
| Reaction<br>temperature | 37°C controlled | 37°C controlled |
| Differences | | |
| Antigen | Human recombinant SS-A/Ro<br>60 (kDa) protein | Purified recombinant Ro60<br>antigen |
| Product Code | LKJ | LLL |
| Sample Dilution<br>(taking a 1%<br>pipetting<br>imprecision into<br>consideration, this<br>sample dilution is<br>regarded as a<br>similarity) | 1:100 | 1:23 |
| | EliA Ro60<br>(Proposed Device) | QUANTA Flash Ro60 |
| Instrumentation | EliA Ro60 uses the EliA IgG<br>method on the instruments<br>Phadia 250 and the E-Modules<br>of the Phadia 2500 and Phadia<br>5000 series. | Quanta Flash Ro60 is run on<br>the BIO-FLASH® instrument. |
| Detection antibody<br>(conjugate) | IgG conjugate: anti-human IgG<br>β-Galactosidase (mouse<br>monoclonal antibodies) | Isoluminol conjugated anti-<br>human IgG |
| Signal | Fluorescence | Relative Light Units (RLU) |
| Calibration | 6-point total IgG Calibration<br>6 vials of human IgG at<br>concentrations of<br>$0 - 4 - 10 - 20 - 100 - 600$ µg/L | Lot specific Master Curve and<br>two Calibrators<br>(Sold separately) |
| Calibration curve | Option to store curve for up to<br>28 days and run curve controls<br>in each assay for calibration | These calibrators are designed<br>for 4 calibrations. The total<br>time the calibrator tubes can be<br>uncapped onboard the<br>instrument is 8 hours. If the<br>calibrators are left uncapped,<br>onboard, for any longer period<br>of time, they should be<br>discarded. Using the same<br>calibrator tubes for more than 8<br>hours can result in improper<br>calibration of the assay, which<br>in turn could give erroneous<br>results. |
| Interpretation of<br>results | Negative < 7 EliA U/mL<br>Equivocal 7-10 EliA U/mL<br>Positive > 10 EliA U/mL | Negative: < 20 CU Positive: > 20 CU |
| Substrate | Development Solution 0.01 %<br>4-Methylumbelliferyl-β-D-<br>galactoside,<br><0.0010% preservative*<br>*Preservative: mixture of 5-<br>chloro-2-methyl-2H-isothiazol-<br>3-one [EC no. 247-500-7] and 2-<br>methyl-2H-isothiazol-3-one [EC<br>no. 220-239-6] (3:1) | "Trigger" reagents |

Phadia AB, Rapsgatan 7P, P.O. Box 6460, 751 37 Uppsala, Sweden

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# SCIENTIF

#### Traditional 510(k) for EliA Ro52 and EliA Ro60, Amendment to K210902 A.7 510(k) Decision Summary_V4

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### Standard/Guidance Document Referenced

- CLSI EP25-A, Evaluation of Stability of In Vitro Diagnostic Reagents, September 2009
- 트 CLSI EP05-A3, Evaluation of Precision of Quantitative Measurement Procedures, September 2014
- 트 CLSI EP06-ED2, Evaluation of the Linearity of Quantitative Measurement Procedures, November 2020
- I CLSI EP07-ED3, Interference Testing in Clinical Chemistry, September 2018
- I CLSI EP37-ED1, Supplemental Tables for Interference Testing, September 2018
- I CLSI EP09c-ED3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples, June 2018
- I CLSI EP17-A2, Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures, June 2012
- 트 CLSI EP28-A3c, Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory, October 2010

# Test Principle

The EliA tests are fluorescence enzyme immunoassays for the detection and measurement of human antibodies based on EliA solid-phase components, which contain specific antigens for the antibodies to be measured.

The specific antigen for the antibodies to be detected is bound to the EliA solid phase component (EliA Well). The EliA wells are molded cups comparable to excised wells from a microtiter plate. They are made of polystyrene and 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.

If present in the patient's specimen, antibodies to these proteins bind to their specific antigen. After washing away non-bound antibodies, enzyme-labeled antibodies against human IgG antibodies (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 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.

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### Performance Characteristics

#### 1. Analytical performance:

- a) Precision/Reproducibility:
To determine the precision of the assay on the Phadia 250 instrument and the Phadia 2500 and Phadia 5000 instrument series, the variability was assessed on 5 samples.

Three lots were used to determine the precision of the assay on Phadia 250 (totaling 252 replicate determinations per sample).

One lot was used to determine the precision of the assay on Phadia 2500E, which is a representative of the Phadia 2500 and Phadia 5000 instrument series.

#### On Phadia 250:

To determine the precision of the assay on the Phadia 250 instrument, the variability was assessed in a study with 21 runs by examining the samples in 252 replicates on 3 Phadia 250 instruments over 7 days. The data was calculated against the calibration curve from Day 1. The statistical evaluation was performed by Analysis of Variance. The results are given in the tables below.

| Mean EliA<br>U/mL | n | Within-Run | | Between-Run | | Between-<br>Instrument | | Total<br>Imprecision | |
|-------------------|-----|------------|-----|-------------|-----|------------------------|-----|----------------------|-----|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 4.4 | 252 | 0.2 | 3.7 | 0.3 | 5.8 | 0.1 | 1.3 | 0.3 | 6.9 |
| 8.3 | 252 | 0.3 | 3.0 | 0.3 | 3.2 | 0.0 | 0.0 | 0.4 | 5.0 |
| 10.6 | 252 | 0.3 | 2.5 | 0.3 | 2.9 | 0.1 | 0.7 | 0.5 | 4.1 |
| 32.8 | 252 | 0.8 | 2.4 | 1.2 | 3.6 | 0.6 | 1.7 | 2.0 | 5.9 |
| 215.4 | 252 | 8.3 | 3.9 | 7.9 | 3.7 | 8.2 | 3.8 | 14.7 | 5.7 |

EliA Ro52 on Phadia 250:

EliA Ro60 on Phadia 250:

| Mean EliA<br>U/mL | n | Within-Run | | Between-Run | | Between-<br>Instrument | | Total<br>Imprecision | |
|-------------------|-----|------------|-----|-------------|-----|------------------------|-----|----------------------|-----|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 3.5 | 252 | 0.1 | 3.6 | 0.3 | 7.2 | 0.1 | 1.5 | 0.3 | 8.2 |
| 6.1 | 252 | 0.2 | 3.0 | 0.3 | 4.5 | 0.0 | 0.0 | 0.3 | 5.4 |
| 10.2 | 252 | 0.2 | 2.3 | 0.3 | 3.2 | 0.1 | 0.7 | 0.5 | 4.4 |
| 32.2 | 252 | 1.1 | 3.3 | 1.3 | 3.9 | 0.0 | 0.0 | 1.9 | 5.8 |
| 201.0 | 252 | 9.6 | 4.8 | 11.6 | 5.7 | 2.9 | 1.4 | 18.6 | 9.1 |

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Within-lab Imprecision:

To determine the within-lab precision of the assay, the variability was assessed in a study with 40 runs by examining the samples in 80 replicates on 1 instrument over 20 days. The data was calculated against the calibration curve from Day 1. The statistical evaluation was performed by Analysis of Variance. The results are given in the table below.

### EliA Ro52:

| Mean<br>(EliA U/mL) | Within-Run | | Within-Lab Imprecision | |
|---------------------|------------|-----|------------------------|-----|
| | SD | %CV | SD | %CV |
| 3.0 | 0.1 | 2.9 | 0.1 | 4.6 |
| 7.9 | 0.2 | 2.9 | 0.3 | 3.9 |
| 10.5 | 0.3 | 2.5 | 0.6 | 5.3 |
| 69.1 | 1.9 | 2.7 | 4.9 | 7.1 |

EliA Ro60:

| Mean<br>(EliA U/mL) | Within-Run | | Within-Lab Imprecision | |
|---------------------|------------|-----|------------------------|-----|
| | SD | %CV | SD | %CV |
| 2.6 | 0.1 | 5.4 | 0.2 | 8.9 |
| 6.3 | 0.2 | 2.6 | 0.6 | 9.7 |
| 10.0 | 0.2 | 2.0 | 0.7 | 6.9 |
| 33.3 | 1.3 | 3.8 | 2.6 | 7.7 |

### Phadia 2500 and Phadia 5000 instrument series:

To determine the precision of the assay on the of the Phadia 2500 and Phadia 5000 instrument series (E-module), the variability was assessed in a study with 21 runs by examining the samples in 84 replicates on 3 Phadia 2500E instruments over 7 days. The data was calculated against the calibration curve from Day 1. The statistical evaluation was performed by Analysis of Variance. The results are given in the table below.

| | | | EliA Ro52 on Phadia 2500 and Phadia 5000 instrument series: | |
|--|--|--|-------------------------------------------------------------|--|
| | | | | |

| Mean | | Within-Run | | Between- | | Between- | | Total | |
|-------|----|------------|-----|----------|-----|------------|-----|-------------|-----|
| EliA | n | | | Run | | Instrument | | Imprecision | |
| U/mL | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 4.7 | 84 | 0.2 | 4.0 | 0.1 | 3.0 | 0.1 | 2.2 | 0.3 | 5.0 |
| 8.6 | 84 | 0.3 | 3.5 | 0.2 | 2.3 | 0.1 | 0.7 | 0.4 | 4.2 |
| 11.0 | 84 | 0.3 | 2.5 | 0.5 | 4.7 | 0.2 | 1.9 | 0.6 | 5.4 |
| 36.3 | 84 | 1.5 | 4.0 | 1.0 | 2.8 | 0.0 | 0.0 | 1.8 | 4.9 |
| 212.9 | 84 | 10.3 | 4.8 | 9.1 | 4.3 | 0.0 | 0.0 | 13.7 | 6.4 |

Phadia AB, Rapsgatan 7P, P.O. Box 6460, 751 37 Uppsala, Sweden

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| Mean | | Within-Run | | Between-<br>Run | | Between-<br>Instrument | | Total<br>Imprecision | |
|--------------|----|------------|-----|-----------------|-----|------------------------|-----|----------------------|-----|
| EliA<br>U/mL | n | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 3.6 | 84 | 0.2 | 5.3 | 0.3 | 6.9 | 0.1 | 1.4 | 0.3 | 8.7 |
| 6.6 | 84 | 0.2 | 2.9 | 0.3 | 4.3 | 0.2 | 2.9 | 0.4 | 5.2 |
| 10.5 | 84 | 0.3 | 2.9 | 0.4 | 3.4 | 0.1 | 0.8 | 0.5 | 4.5 |
| 35.8 | 84 | 1.2 | 3.3 | 1.3 | 3.7 | 0.0 | 0.0 | 1.8 | 4.9 |
| 219.9 | 84 | 11.7 | 5.3 | 10.3 | 4.7 | 10.7 | 4.9 | 18.9 | 7.1 |

EliA Ro60 on Phadia 2500 and Phadia 5000 instrument series:

### b) Linearity/Assay Reportable Range:

The linearity of EliA Ro52 and EliA Ro60 was evaluated by a study performed according to CLSI EP06-ED2. Three patient serum samples were serially diluted in at least 9 dilution steps using a blood donor serum sample and tested in quadruplicates on one batch EliA Ro52 Well and one batch EliA Ro60 Well with one set of system reagents on the Phadia 250 instrument and the Phadia 2500E instrument, respectively. Results were analyzed according to the guideline performing a weighted linear regression with intercept. All sample sets showed dilution linearity for the entire measuring range.

| Sample | Parameter | Estimate | Lower<br>95% CI | Upper<br>95% CI | R2 |
|------------------------|-----------|----------|-----------------|-----------------|--------|
| 1 | Intercept | 1.226 | 0.865 | 1.587 | 0.9989 |
| 1 | slope | 0.987 | 0.961 | 1.012 | 0.9989 |
| 2 | Intercept | 0.435 | 0.320 | 0.551 | 0.9994 |
| 2 | slope | 0.994 | 0.977 | 1.011 | 0.9994 |
| 3 | Intercept | 0.091 | -0.053 | 0.235 | 0.9899 |
| 3 | slope | 0.948 | 0.884 | 1.013 | 0.9899 |
| Sample 1-3<br>combined | Intercept | 0.025 | -0.080 | 0.130 | 0.9928 |
| Sample 1-3<br>combined | slope | 1.014 | 0.983 | 1.045 | 0.9928 |

EliA Ro52 on Phadia 250:

EliA Ro52 on Phadia 2500E:

| Sample | Parameter | Estimate | Lower<br>95% CI | Upper<br>95% CI | R2 |
|--------|-----------|----------|-----------------|-----------------|--------|
| 1 | Intercept | 0.776 | -0.291 | 1.842 | 0.9941 |
| | slope | 0.862 | 0.811 | 0.913 | |
| 2 | Intercept | 0.490 | 0.265 | 0.715 | 0.9980 |
| | slope | 0.911 | 0.884 | 0.938 | |

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| 3 | Intercept | -0.045 | -0.163 | 0.073 | 0.9875 |
|------------|-----------|--------|--------|-------|--------|
| | slope | 0.904 | 0.831 | 0.976 | |
| Sample 1-3 | Intercept | -0.010 | -0.114 | 0.094 | |
| combined | slope | 0.928 | 0.902 | 0.953 | 0.9942 |

For EliA Ro52, linearity was shown for the entire measuring range.

EliA Ro60 on Phadia 250:

| Sample | Parameter | Estimate | Lower<br>95% CI | Upper<br>95% CI | R2 |
|------------------------|-----------|----------|-----------------|-----------------|--------|
| 1 | Intercept | 0.674 | 0.397 | 0.951 | 0.9961 |
| | slope | 1.016 | 0.967 | 1.064 | 0.9961 |
| 2 | Intercept | 1.282 | 0.515 | 2.048 | 0.9996 |
| | slope | 1.047 | 1.030 | 1.064 | 0.9996 |
| 3 | Intercept | 0.053 | 0.020 | 0.086 | 0.9994 |
| | slope | 0.988 | 0.971 | 1.005 | 0.9994 |
| Sample 1-3<br>combined | Intercept | -0.004 | -0.076 | 0.069 | 0.9979 |
| | slope | 1.063 | 1.044 | 1.081 | 0.9979 |

EliA Ro60 on Phadia 2500E:

| Sample | Parameter | Estimate | Lower<br>95% CI | Upper<br>95% CI | R2 |
|------------------------|-----------|----------|-----------------|-----------------|--------|
| 1 | Intercept | 0.136 | -0.125 | 0.396 | 0.9967 |
| | slope | 1.013 | 0.968 | 1.058 | 0.9967 |
| 2 | Intercept | 0.910 | 0.171 | 1.649 | 0.9949 |
| | slope | 1.033 | 0.980 | 1.086 | 0.9949 |
| 3 | Intercept | 0.040 | 0.009 | 0.071 | 0.9992 |
| | slope | 0.963 | 0.942 | 0.984 | 0.9992 |
| Sample 1-3<br>combined | Intercept | -0.039 | -0.131 | 0.053 | 0.9964 |
| | slope | 1.069 | 1.045 | 1.093 | 0.9964 |

For EliA Ro60, linearity was shown for the entire measuring range.

### Hook Effect/Over the Range Results:

Not applicable. Results above the upper limit of the measuring range are reported as ">240" for both assays. No recommendations are made for dilution of samples outside measuring range in the Directions for Use.

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#### c) Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

#### Traceability:

The IgG calibrators are traceable (via unbroken chain of calibrations) to the International Reference Preparation (IRP) 67/86 of Human Serum Immunoglobulins A. G and M from WHO. New batches of IgG calibrators are compared to a secondary standard (standardized with the IRP) or the IRP directly and adjusted accordingly to meet the correct concentration.

The instrument measures specific IgG concentrations in ug/L. By using a conversion factor given by the lot-specific code of the EliA test well. the results are automatically converted to EliA U/mL.

#### Stabilitv:

Data for open and closed real-time stability and on-board stability of EliA IgG reagents and general EliA reagents on Phadia 250 as well as on the E-module of the Phadia 2500 and Phadia 5000 series were already cleared with several other EliA tests, e.g., under K141375 (EliA M2 on Phadia 250). For the Phadia 2500 and Phadia 5000 instrument series, they were already cleared under K061165/A003 (EliA CCP).

### Shelf-life:

The stability of EliA Ro52 and EliA Ro60 Wells was evaluated with a real-time study. The results support stability of the test under the recommended storage of 2 – 8°C for up to 24 months for EliA Ro52 and 18 months for EliA Ro60.

### On-board stability:

The on-board stability EliA Ro52 and EliA Ro60 carriers (containing the antigen coated wells) was tested over 8 weeks using 3 positive and 2 negative samples only on the Phadia 250 instrument. As the storage conditions in the E-module of the Phadia 2500 and Phadia 5000 series are similar to the Phadia 250, the results can also be used for stability claims for these instruments. The on-board stability for the Phadia 250 was determined to be 28 days at 2-8°C.

### Open Stability:

Stability after first opening of the foil bag containing the EliA Ro52 and EliA Ro60 wells was tested with a real-time study. According to the accelerated stability study, a shelf-life of 9 months at 2-8°C after first opening can be assigned to EliA Ro52 and EliA Ro60 wells.

### d) Detection Limit:

Four blank and four low level samples were measured with two different reagent sets (two lots of antigen wells). The four blank samples were created from depleted IgG sera, each diluted with EliA Sample Diluent. The blank samples and the low-

{19}------------------------------------------------

> level samples were assayed in three runs using two different sets of EliA Ro52 and EliA Ro60 Well lots over three different days on a Phadia 250 and Phadia 2500E each in 5-fold determination. For each instrument type, the total number of combined observations for blank and low-level samples is 120 (60 per reagent set, 15 per sample and reagent set).

The results are summarized in the tables below.

EliA Ro52:

| Instrument | LoB | LoD | LoQ |
|-------------------------------------------------------|-----------|-----------|-----------|
| | EliA U/mL | EliA U/mL | EliA U/mL |
| Phadia 250 | 0.0 | 0.2 | 0.6 |
| E-module of the Phadia 2500 and<br>Phadia 5000 series | 0.1 | 0.3 | 0.7 |

The LoD for EliA Ro52 is 0.3 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 (B) less than 5%; based on 240 determinations with 120 blank and 120 low-level replicates per instrument type; and LoB of 0.1 EliA U/mL.

A harmonized LoB of 0.1 EliA U/mL, LoD of 0.3 EliA U/mL, and LoQ of 0.7 EliA U/mL for the immunoassay was used.

EliA Ro60:

| Instrument | LoB<br>EliA U/mL | LoD<br>EliA U/mL | LoQ<br>EliA U/mL |
|-------------------------------------------------------|------------------|------------------|------------------|
| Phadia 250 | 0.1 | 0.2 | 0.4 |
| E-module of the Phadia 2500 and<br>Phadia 5000 series | 0.1 | 0.2 | 0.5 |

The LoD for EliA Ro60 is 0.4 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 (ß) less than 5%; based on 240 determinations with 120 blank and 120 low-level replicates per instrument type; and LoB of 0.1 EliA U/mL.

As the LoB, LoD and LoQ were harmonized over both instrument types, the LoB was set to 0.1 EliA U/mL, the LoD to 0.2 EliA U/mL, and the LoQ to 0.5 EliA U/mL. The DfU states our current worldwide harmonized LoD of 0.4 EliA U/mL for both instrument types as the lower limit of the measuring range.

{20}------------------------------------------------

#### e) Analytical specificity:

#### Endogenous and Exogenous Interference:

A study was run to investigate whether high concentrations of potentially interfering substances in serum, like bilirubin, hemoglobin, lipemic factor, rheumatoid factor, human IgG, Ibuprofen, Losartan, Hydroxychloroquine, Azathioprine, Prednisone, Rituximab, Infliximab, Diltiazem and Omeprazole adversely affect the results of the new device.

Three serum samples (one negative sample, one sample with a concentration within the equivocal range, and one high positive sample) were spiked with the different interfering substances or blank solution. The samples were tested in triplicates. A calibration curve was run in duplicate. The runs were repeated twice. One batch of EliA antigen wells and one batch of system reagents were used throughout the studies.

The ratio of blank/spiked sample ranged from 0.90 – 1.10 for EliA Ro52 and EliA Ro60. No interference was observed up to the concen…