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 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.

AI/ML Overview

The provided document describes the EliA Ro52 and EliA Ro60 Immunoassays, which are intended for the semi-quantitative measurement of IgG antibodies directed to Ro52 and Ro60 in human serum, respectively. These devices aid in the diagnosis of specific autoimmune diseases. The document includes detailed analytical and clinical performance data to support the substantial equivalence claim.

Here's an analysis of the acceptance criteria and the study that proves the device meets them:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state formal "acceptance criteria" in a tabulated format. Instead, it presents various analytical and clinical performance metrics, implying that these metrics, when compared to the predicate device and established clinical utility, are deemed acceptable for the device's intended use. Based on the provided performance data, the implicit acceptance criteria would likely be related to aspects like precision, linearity, detection limits, specificity (interference), and clinical sensitivity/specificity.

Below is a table summarizing key performance indicators reported in the document:

Table 1: Key Performance Metrics for EliA Ro52 and EliA Ro60

Performance Metric	EliA Ro52 Reported Performance	EliA Ro60 Reported Performance	Implied Acceptance Criterion (General, not prescriptive)
Precision (Total Imprecision)	On Phadia 250: %CV 4.1% - 6.9% On Phadia 2500/5000: %CV 4.2% - 6.4%	On Phadia 250: %CV 4.4% - 9.1% On Phadia 2500/5000: %CV 4.5% - 7.1%	Total imprecision (Total %CV) should be within acceptable limits for diagnostic assays, demonstrating consistency and reliability across runs, instruments, and days.
Linearity (R² value)	Phadia 250: 0.9899 - 0.9994 (single), 0.9928 (combined) Phadia 2500E: 0.9875 - 0.9980 (single)	Phadia 250: 0.9961 - 0.9996 (single), 0.9979 (combined) Phadia 2500E: 0.9949 - 0.9992 (single)	The assay should demonstrate linearity across its entire measuring range, indicated by a high R² value (close to 1) for regression analysis of serially diluted samples.
Detection Limit (LoD)	0.3 EliA U/mL	DfU states 0.4 EliA U/mL (harmonized)	The Limit of Detection (LoD) should be sufficiently low to detect clinically relevant concentrations of the analyte.
Analytical Specificity	No interference observed from listed endogenous/exogenous substances.	No interference observed from listed endogenous/exogenous substances.	The assay should not be significantly affected by common interfering substances (e.g., bilirubin, hemoglobin, rheumatoid factor, common medications) at clinically relevant concentrations.
Method Comparison (vs. Predicate)	EliA Ro52 vs. QUANTA Flash Ro52: PPA 80.8% - 92.3%, NPA 90.7% - 98.4%, TPA 91.2% - 93.4%	EliA Ro60 vs. QUANTA Flash Ro60: PPA 93.9% - 97.0%, NPA 81.6% - 92.1%, TPA 91.3% - 93.3%	Agreement with the legally marketed predicate device (expressed as Positive Percent Agreement (PPA), Negative Percent Agreement (NPA), and Total Percent Agreement (TPA)) should be high, demonstrating comparable performance.
Clinical Sensitivity (EliA Ro52)	SLE: 47.5% - 50.8% SS: 50.0% - 55.0% IIM: 36.2% - 37.2% SSc: 20.9% - 26.4%	N/A (Ro52 specific)	Clinical sensitivity for the specified autoimmune diseases (Sjögren's syndrome, systemic lupus erythematosus, systemic sclerosis, idiopathic inflammatory myopathies) should be clinically acceptable and comparable to established diagnostic benchmarks for similar assays. Specific numerical cut-offs are implied by the data presented.
Clinical Specificity (EliA Ro52)	SLE control: 95.6% - 96.9% SS control: 95.6% - 96.9% IIM control: 95.6% - 96.9% SSc control: 95.6% - 96.9%	N/A (Ro52 specific)	Clinical specificity against control groups (other autoimmune diseases, infectious diseases) should be high to minimize false positives.
Clinical Sensitivity (EliA Ro60)	N/A (Ro60 specific)	SLE: 48.3% - 50.8% SS: 68.3% - 71.7%	Clinical sensitivity for the specified autoimmune diseases (Sjögren's syndrome, systemic lupus erythematosus) should be clinically acceptable and comparable to established diagnostic benchmarks for similar assays. Specific numerical cut-offs are implied by the data presented.
Clinical Specificity (EliA Ro60)	N/A (Ro60 specific)	SLE control: 98.4% - 98.6% SS control: 98.4% - 98.6%	Clinical specificity against control groups (other autoimmune diseases, infectious diseases) should be high to minimize false positives.

The acceptance of these performance metrics is implicitly based on comparison with the predicate devices (QUANTA Flash Ro52 and QUANTA Flash Ro60) and the established clinical utility in diagnosing the mentioned autoimmune diseases. The FDA's substantial equivalence determination (K210902) confirms that the submitted data supports the claim that the new devices perform comparably to the predicate devices.

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

The document describes several test sets for different performance characteristics:

Precision/Reproducibility:
- Phadia 250: 5 samples, each with 252 replicate determinations (across 3 instruments, 7 days, 21 runs).
- Within-lab Imprecision: Samples in 80 replicates on 1 instrument over 20 days (40 runs).
- Phadia 2500 and Phadia 5000 series (E-module): 5 samples, each with 84 replicates (across 3 instruments, 7 days, 21 runs).
Linearity/Assay Reportable Range: 3 patient serum samples serially diluted in at least 9 dilution steps, tested in quadruplicates.
Detection Limit: 4 blank and 4 low-level samples, tested in 5-fold determination, across 2 different reagent sets (totaling 120 combined observations for blank and low-level per instrument type).
Analytical Specificity (Interference): 3 serum samples (negative, equivocal, high positive), spiked with interfering substances, tested in triplicates.
Method Comparison with Predicate Device:
- EliA Ro52: 208 patient serum samples (only 181 used in agreement calculations due to samples outside measuring range).
- EliA Ro60: 208 patient serum samples (only 104 used in agreement calculations due to samples outside measuring range).
Clinical Sensitivity and Specificity:
- EliA Ro52: 755 clinically and ethnically defined serum samples. These include:
  - Diagnostic group: SLE (n=120), Sjögren's syndrome (n=60), IIM (n=94), SSc (n=91) - total 365.
  - Disease control group: 390 samples (various autoimmune and infectious diseases).
- EliA Ro60: 713 clinically and ethnically defined serum samples. These include:
  - Diagnostic group: SLE (n=120), Sjögren's syndrome (n=60) - total 180.
  - Disease control group: 533 samples (various autoimmune and infectious diseases, excluding SSc).

Data Provenance:
The document states that the clinical samples used for sensitivity and specificity determination were from "clinically and ethnically defined serum samples, including those of US origin." This indicates a mix of retrospective (clinically defined, likely banked samples) and potentially prospective (if US origin implies some form of fresh collection for the study) data. It is not explicitly stated whether it was purely retrospective or involved prospective collections, but the description "clinically and ethnically defined serum samples" strongly suggests retrospective use of samples with established diagnoses.

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

The document does not provide information on the number of experts used to establish the ground truth for the test sets (e.g., for diagnoses of SLE, SS, IIM, SSc). It only states that samples were "clinically and ethnically defined." This typically implies that patient diagnoses were established through standard clinical practice by medical professionals, but the specifics of these experts' qualifications or the consensus process are not detailed in this submission summary.

4. Adjudication Method for the Test Set

The document does not specify an adjudication method for establishing the ground truth for the test set. It refers to diagnoses as "clinically defined," which generally suggests diagnosis was made by treating physicians following clinical guidelines, but no adjudication process like 2+1 or 3+1 by independent experts is mentioned for the study.

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

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This document describes an in vitro diagnostic (IVD) device, specifically an immunoassay for measuring antibody levels. MRMC studies are typically performed for imaging devices or other diagnostic tools where human interpretation is involved and needs to be compared with and/or augmented by AI. This device is an automated laboratory test, and its performance is evaluated directly through analytical measurements and comparison to a predicate device, as well as clinical sensitivity and specificity against established clinical diagnoses.

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

Yes, this is a standalone device. The EliA Ro52 and EliA Ro60 Immunoassays are "automated semi-quantitative solid phase fluoroenzymeimmunoassays" run on "Phadia 250 instrument and the Phadia 5000 instrument series (E-modules)." The results are automatically converted to EliA U/mL. The performance characteristics described (precision, linearity, detection limit, analytical specificity, clinical sensitivity, specificity) are all based on the output of the automated instrument and reagents, independent of human interpretation during the actual test process itself. The interpretation of results (negative, equivocal, positive) is based on defined cut-off values.

7. The Type of Ground Truth Used (Expert Consensus, Pathology, Outcomes Data, etc.)

For the clinical sensitivity and specificity studies, the ground truth was clinical diagnosis. The samples were "clinically and ethnically defined serum samples" with established diagnoses of various autoimmune diseases (SLE, SS, IIM, SSc) and control conditions. This implies that the diagnosis was based on the standard clinical criteria and medical records established by healthcare professionals, rather than a separate expert consensus panel or specific pathology results for the study.

8. The Sample Size for the Training Set

The document does not describe a "training set" in the context of machine learning or AI models, as this is an immunoassay and not an AI/ML-based device.

However, if "training set" is generally interpreted as samples used for assay development, optimization, or establishing parameters like cut-offs, the relevant information is:

Cut-off definition: A cohort of 69 healthy blood donors, 19 SLE patients, and 9 Sjögren's syndrome (SS) patients for EliA Ro52, and 70 healthy blood donors, 22 SLE patients, and 6 Sjögren's syndrome patients for EliA Ro60, were used to define the cut-off values. This can be considered the 'development' or 'calibration' set for determining the interpretive thresholds.

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

As noted above, this device does not utilize a machine learning "training set" in the conventional sense. For the samples used to establish the cut-off values (which could be considered analogous to determining a 'decision boundary' in an ML context), the ground truth was established based on the clinical diagnoses of the patient samples (healthy blood donors, SLE patients, Sjögren's syndrome patients). The document states that the initial cut-off for EliA Ro52, set using SLE and SS patients, was later verified with additional IIM and SSc patient sera. This indicates clinical diagnosis as the gold standard.

Summary

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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 (DICE@fda.hhs.gov) or phone (1-800-638-2041 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)

Type of Use (Select one or both, as applicable)
☑ Prescription Use (Part 21 CFR 801 Subpart D)	☐ Over-The-Counter Use (21 CFR 801 Subpart C)

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 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."

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Indications for Use

510(k) Number (if known)

K210902

Device Name

EliA Ro60

Indications for Use (Describe)

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

Prescription Use (Part 21 CFR 801 Subpart D)	☑
Over-The-Counter Use (21 CFR 801 Subpart C)	☐

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.

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."

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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 ABRapsgatan 7PP.O. Box 6460751 37 Uppsala, Sweden
Distributor:	Phadia US Inc.4169 Commercial AvenuePortage, MI 49002
Company Contact Person:	Jane AnthonySenior Manager, Quality Systems and CompliancePhadia US Inc.4169 Commercial Avenue, Portage, MI 49002269-254-6833jane.anthony@thermofisher.com

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 TestSystem
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:

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:

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(Proposed Device)	QUANTA Flash Ro52

Similarities
Assay Intended Use	EliA Ro52 is intended for the invitro semi-quantitativemeasurement of IgG antibodiesdirected to Ro52 in humanserum as an aid in the diagnosisof Sjögren's syndrome (SS),systemic lupus erythematosus(SLE), systemic sclerosis (SSc)and idiopathic inflammatorymyopathies (IIM) in conjunctionwith other laboratory andclinical findings. EliA Ro52uses the EliA IgG method.	QUANTA Flash® Ro52 is achemiluminescentimmunoassay for the semi-quantitative determination ofIgG anti-Ro52 autoantibodiesin human serum. The presenceof anti-Ro52 autoantibodies, inconjunction with clinicalfindings and other laboratorytests, is an aid in the diagnosisof Systemic LupusErythematosus (SLE),Sjögren's Syndrome, SystemicSclerosis, IdiopathicInflammatory Myopathies.
Classification	Class II	Class II
Regulation Number	866.5100	866.5100
Internal Controls	Positive and negative Controlprovided with EliA ANA 3Positive Control 250 /2500/5000 and EliAIgG/IgM/IgA Negative Control250/2500/5000, respectively.	QUANTA Flash® Ro52 Controls(Negative and Positive Control)
Assay technique	ELISA	ELISA
Type of test	Semi-quantitative	Semi-quantitative
Reactiontemperature	37°C controlled	37°C controlled
Differences
Antigen	Human recombinant SS-A/Ro52 (kDa) protein	Purified recombinant Ro52antigen
Product Code	LKJ	OBE
Sample Dilution(taking a 1%pipettingimprecision intoconsideration, thissample dilution isregarded as a	1:100	1:23
	EliA Ro52(Proposed Device)	QUANTA Flash Ro52
Reporting of results	EliA U/mL (arbitrary)	Chemiluminescent units (CU)
Instrumentation	EliA Ro52 uses the EliA IgGmethod on the instrumentsPhadia 250 and the E-Modulesof the Phadia 2500 and Phadia5000 series.	Quanta Flash Ro52 is run onthe BIO-FLASH® instrument.
Detection antibody(conjugate)	IgG conjugate: anti-human IgGß-Galactosidase (mousemonoclonal antibodies)	Isoluminol conjugated anti-human IgG
Signal	Fluorescence	Relative Light Units (RLU)
Calibration	6-point total IgG Calibration6 vials of human IgG atconcentrations of0–4–10–20–100–600 µg/L	Lot specific Master Curve andtwo Calibrators(Sold separately)
Calibration curve	Option to store curve for up to28 days and run curve controlsin each assay for calibration	These calibrators are designedfor 4 calibrations. The totaltime the calibrator tubes can beuncapped onboard theinstrument is 8 hours. If thecalibrators are left uncapped,onboard, for any longer periodof time, they should bediscarded. Using the samecalibrator tubes for more than 8hours can result in impropercalibration of the assay, whichin turn could give erroneousresults.
Interpretation ofresults	Negative < 7 EliA U/mLEquivocal 7-10 EliA U/mLPositive > 10 EliA U/mL	Negative: < 20 CU Positive: > 20CU
Substrate	Development Solution 0.01 %4-Methylumbelliferyl-ß-D-galactoside,<0.0010% preservative**Preservative: mixture of 5-chloro-2-methyl-2H-isothiazol-3-one [EC no. 247-500-7] and 2-methyl-2H-isothiazol-3-one [ECno. 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(Proposed Device)	QUANTA Flash Ro60
Similarities
Assay Intended Use	EliA Ro60 is intended for the in vitro semi-quantitativemeasurement of IgG antibodiesdirected to Ro60 in humanserum as an aid in the diagnosisof Sjögren's syndrome (SS) andsystemic lupus erythematosus(SLE) in conjunction with otherlaboratory and clinical findings.EliA Ro60 uses the EliA IgGmethod.	QUANTA Flash Ro60 is achemiluminescentimmunoassay for the semi-quantitative determination ofIgG anti-Ro60 autoantibodiesin human serum. The presenceof anti-Ro60 autoantibodies, inconjunction with clinicalfindings and other laboratorytests, is an aid in the diagnosisof Systemic LupusErythematosus and Sjögren'sSyndrome.
Classification	Class II	Class II
Regulation Number	866.5100	866.5100
Internal Controls	Positive and negative Controlprovided with EliA ANA 3Positive Control 250 /2500/5000 and EliAIgG/IgM/IgA Negative Control250 / 2500/5000, resp.	QUANTA Flash® Ro60Controls (Negative and PositiveControl)
Assay technique	ELISA	ELISA
Type of test	Semi-quantitative	Semi-quantitative
Reactiontemperature	37°C controlled	37°C controlled
Differences
Antigen	Human recombinant SS-A/Ro60 (kDa) protein	Purified recombinant Ro60antigen
Product Code	LKJ	LLL
Sample Dilution(taking a 1%pipettingimprecision intoconsideration, thissample dilution isregarded as asimilarity)	1:100	1:23
	EliA Ro60(Proposed Device)	QUANTA Flash Ro60
Instrumentation	EliA Ro60 uses the EliA IgGmethod on the instrumentsPhadia 250 and the E-Modulesof the Phadia 2500 and Phadia5000 series.	Quanta Flash Ro60 is run onthe BIO-FLASH® instrument.
Detection antibody(conjugate)	IgG conjugate: anti-human IgGβ-Galactosidase (mousemonoclonal antibodies)	Isoluminol conjugated anti-human IgG
Signal	Fluorescence	Relative Light Units (RLU)
Calibration	6-point total IgG Calibration6 vials of human IgG atconcentrations of$0 - 4 - 10 - 20 - 100 - 600$ µg/L	Lot specific Master Curve andtwo Calibrators(Sold separately)
Calibration curve	Option to store curve for up to28 days and run curve controlsin each assay for calibration	These calibrators are designedfor 4 calibrations. The totaltime the calibrator tubes can beuncapped onboard theinstrument is 8 hours. If thecalibrators are left uncapped,onboard, for any longer periodof time, they should bediscarded. Using the samecalibrator tubes for more than 8hours can result in impropercalibration of the assay, whichin turn could give erroneousresults.
Interpretation ofresults	Negative < 7 EliA U/mLEquivocal 7-10 EliA U/mLPositive > 10 EliA U/mL	Negative: < 20 CU Positive: > 20 CU
Substrate	Development Solution 0.01 %4-Methylumbelliferyl-β-D-galactoside,<0.0010% preservative**Preservative: mixture of 5-chloro-2-methyl-2H-isothiazol-3-one [EC no. 247-500-7] and 2-methyl-2H-isothiazol-3-one [ECno. 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 EliAU/mL	n	Within-Run		Between-Run		Between-Instrument		TotalImprecision
		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 EliAU/mL	n	Within-Run		Between-Run		Between-Instrument		TotalImprecision
		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(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(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-Run		Between-Instrument		TotalImprecision
EliAU/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	Lower95% CI	Upper95% 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 1-3combined	Intercept	0.025	-0.080	0.130	0.9928
Sample 1-3combined	slope	1.014	0.983	1.045	0.9928

EliA Ro52 on Phadia 250:

EliA Ro52 on Phadia 2500E:

Sample	Parameter	Estimate	Lower95% CI	Upper95% 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	Lower95% CI	Upper95% 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 1-3combined	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	Lower95% CI	Upper95% 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 1-3combined	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-

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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 andPhadia 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	LoBEliA U/mL	LoDEliA U/mL	LoQEliA U/mL
Phadia 250	0.1	0.2	0.4
E-module of the Phadia 2500 andPhadia 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.

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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 concentrations listed in the table below:

Potential Interfering	Concentration in
Compound	undiluted sample
Bilirubin F	40 mg/dL
Bilirubin C	40 mg/dL
Hemoglobin	1000 mg/dL
Lipemic factor	2000 mg/dL
Rheumatoid factor	500 IU/mL
Human IgG	3500 mg/dL
Ibuprofen	21.9 mg/dL
Losartan	1.14 mg/dL
Hydroxychloroquine	0.23 mg/dL
Azathioprine	0.26 mg/dL
Prednisone	0.01 mg/dL
Rituximab	109 mg/dL
Infliximab	26.4 mg/dL
Omeprazole	0.84 mg/dL
Diltiazem	0.09 mg/dL

Reference Sera:

The panel of CDC ANA human reference sera #1 - #12 was tested with EliA Ro52 and EliA Ro60.

Confidential Page 15 (26)

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In the EliA Ro52 test, CDC ANA #2 (speckled pattern in FANA), CDC ANA #7 (SS-A/Ro) and CDC ANA #10 (Jo-1) showed positive results with 13.9 EliA U/mL, 19.5 EliA U/mL and 247.4 EliA U/mL, respectively. CDC ANA #3 (RNP-Sm, SS-A/Ro, SS-B/La) showed an equivocal result with 7.4 EliA U/mL. All other CDC ANA samples remained negative.

In the EliA Ro60 test, CDC ANA #2 (speckled pattern in FANA), CDC ANA #3 (RNP-Sm, SS-A/Ro, SS-B/La), and CDC ANA #7 (SS-A/Ro) showed positive results with 143.0 EliA U/mL, 146.2 EliA U/mL and 207.2 EliA U/mL, respectively. All other CDC ANA samples remained negative.

f) Assay Cut-Off:

EliA Ro52:

To define the cut-off, a study was performed using a cohort consisting of 69 apparently healthy blood donors, 19 samples from SLE patients and 9 samples from Sjögren's syndrome (SS) patients. The samples were measured on a Phadia 250 instrument.

The cut-off value of EliA Ro52 that was initially set with the intended use target diseases SLE and SS could be verified for the additional intended use groups using 10 IIM and 14 SSc patient sera.

EliA Ro60:

To define the cut-off, a study was performed using a cohort consisting of 70 apparently healthy blood donors, 22 samples from SLE patients and 6 samples from Sjögren's syndrome patients. The samples were measured on a Phadia 250 instrument

The cut-off was set as follows for EliA Ro52 and EliA Ro60:

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

In case of equivocal results, it is recommended to retest the patient after 8-12 weeks.

2. Comparison Studies:

a) Method Comparison with Predicate Device:
A total of 208 patient serum samples with concentrations covering the measuring range were tested with EliA Ro52, EliA Ro60 and QUANTA Flash Ro52 and QUANTA Flash Ro60 assays. Observed concentrations of samples outside the measuring intervals of the EliA and/or QUANTA Flash tests were disregarded in the agreement calculations.

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The tests were run in single determination and evaluated according to their Directions for Use. The results are summarized in the tables below:

n = 181	QUANTA Flash Ro52positive: ≥ 20 CU	QUANTA Flash Ro52negative: < 20 CU	Total
EliA Ro52positive:> 10 EliA U/mL	42	2	44
EliA Ro52negative:< 10 EliA U/mL	10	127	137
Total	52	129	181

EliA Ro52: equivocal results considered negative

	Calculation	Agreement (%)	95% CI
PPA	$100% x 42 / 52$	80.8	67.5 - 90.4
NPA	$100% x 127 / 129$	98.4	94.5 - 99.8
TPA	$100% x (42+127) / 181$	93.4	88.8 - 96.5

EliA Ro52: equivocal results considered positive

n = 181	QUANTA Flash Ro52positive: ≥ 20 CU	QUANTA Flash Ro52negative: < 20 CU	Total
EliA Ro52positive: > 7 EliA U/mL	48	12	60
EliA Ro52negative: < 7 EliA U/mL	4	117	121
Total	52	129	181

	Calculation	Agreement (%)	95% CI
PPA	$100% \times 48 / 52$	92.3	81.5 – 97.9
NPA	$100% 117 / 129$	90.7	84.3 – 95.1
TPA	$100% \times (48+117) / 181$	91.2	86.1 – 94.9

EliA Ro60: equivocal results considered negative

n = 104	QUANTA Flash Ro60 positive: ≥ 20 CU	QUANTA Flash Ro60 negative: < 20 CU	Total
EliA Ro60 positive: > 10 EliA U/mL	62	3	65
EliA Ro60 negative: < 10 EliA U/mL	4	35	39
Total	66	38	104

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	Calculation	Agreement (%)	95% CI
PPA	$100% x 62 / 66$	93.9	85.2 - 98.3
NPA	$100% x 35 / 38$	92.1	78.6 - 98.3
TPA	$100% x (62+35) / 104$	93.3	86.6 - 97.3

EliA Ro60: equivocal results considered positive

n = 104	QUANTA Flash Ro60positive: ≥ 20 CU	QUANTA Flash Ro60negative: < 20 CU	Total
EliA Ro60positive: > 7 EliA U/mL	64	7	71
EliA Ro60negative: < 7 EliA U/mL	2	31	33
Total	66	38	104

	Calculation	Agreement (%)	95% CI
PPA	$100% x 64 / 66$	97.0	89.5 - 99.6
NPA	$100% 31 / 38$	81.6	65.7 - 92.3
TPA	$100% x (64+31) / 104$	91.3	84.2 - 96.0

b) Instrument Comparison:

EliA Ro52:

Performance of EliA Ro52 was evaluated on the Phadia 250 and Phadia 2500E instrument using 57 positive, 6 equivocal and 30 negative samples. The samples were analyzed in single determination on one Phadia 250 and one Phadia 2500E instrument each. The regression analysis results are summarized as follows:

	Intercept	Slope
Estimate	0.91	0.94
95% CI	0.46 - 1.14	0.93 - 0.97

EliA Ro60:

Performance of EliA Ro60 was evaluated on the Phadia 250 and Phadia 2500E instrument using 42 positive, 9 equivocal and 39 negative samples. The samples were analyzed in single determination on one Phadia 250 and one Phadia 2500E instrument each. The regression analysis results are summarized as follows:

	Intercept	Slope
Estimate	0.24	1.01
95% CI	0.15 - 0.48	0.97 – 1.04

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3. Clinical Studies

c) Clinical Sensitivity and Specificity:

EliA Ro52:

In total, 755 clinically and ethnically defined serum samples, including those of US origin, were used to determine sensitivity and specificity of the assay. Samples with a diagnosis of systemic lupus erythematosus (SLE), Sjögren's syndrome (SS), idiopathic inflammatory myopathies (IIM) and systemic sclerosis represent the diagnostic group (target diseases: SLE n=120, Sjögren's syndrome n=60, IIM n=94, SSc n=91).

Samples with various autoimmune and infectious disease diagnoses represent the disease control group (n=390). The results are summarized in the tables below.

Results of clinically defined samples. SLE samples represented the "target disease". SS, IIM and SSc samples were excluded, other autoimmune diseases and infectious diseases were used as "control disease". Equivocal results were either judged as negative or positive.

	Equivocal = negative
Result	SLE	Controls	Total
Positive	57	12	69
Negative	63	378	441
Total	120	390	510

	Value	95% CI
Sensitivity	47.5%	38.3 - 56.8
Specificity	96.9%	94.7 - 98.4

Result	SLE	Controls	Total
Positive	61	17	78
Negative	59	373	432
Total	120	390	510

	Value	95% CI
Sensitivity	50.8%	41.6 - 60.1
Specificity	95.6%	93.1 - 97.4

Results of clinically defined samples. Sjögren's syndrome samples represented the "target disease", SLE, IIM and SSc samples were excluded, other autoimmune diseases and infectious diseases were used as "control disease". Equivocal results were either judged as negative or positive.

Equivocal = negative				Equivocal = positive
Result	SS	Controls	Total	Result	SS	Controls	Total
Positive	30	12	42	Positive	33	17	50
Negative	30	378	408	Negative	27	373	400
Total	60	390	450	Total	60	390	450

SensitivitySensitivity55.0%50.0%36.8 - 63.241.6 - 67.9		Value	95% CI		Value	95% CI

	Specificity	96.9%	94.7 - 98.4	Specificity	95.6%	93.1 - 97.4

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Results of clinically defined samples. IIM samples represented the "target disease", SLE, SS, and SSc samples were excluded, other autoimmune diseases and infectious diseases were used as "control disease". Equivocal results were either judged as negative or positive.

Equivocal = negative
Result	IIM	Controls	Total
Positive	34	12	46
Negative	60	378	438
Total	94	390	484

Equivocal = positive
Result	IIM	Controls	Total
Positive	35	17	52
Negative	59	373	432
Total	94	390	484

	Value	95% CI
Sensitivity	36.2%	26.5 - 46.7
Specificity	96.9%	94.7 - 98.4

	Value	95% CI
Sensitivity	37.2%	27.5 - 47.8
Specificity	95.6%	93.1 - 97.4

Results of clinically defined samples. SSc samples represented the "target disease", SLE, SS and IIM samples were excluded, other autoimmune diseases and infectious diseases were used as "control disease". Equivocal results were either judged as negative or positive.

Equivocal = negative
CC	C	A	T

Result	SSC	Controls	Total
Positive	19	12	31
Negative	72	378	450
Total	91	390	481

	Equivocal = positive
--	----------------------

Result	SSc	Controls	Total
Positive	24	17	41
Negative	67	373	440
Total	91	390	481

	Value	95% CI
Sensitivity	20.9%	13.1 - 30.7
Specificity	96.9%	94.7 - 98.4

	Value	95% CI
Sensitivity	26.4%	17.7 - 36.7
Specificity	95.6%	93.1 - 97.4

The table below shows the results for each clinical subgroup:

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SCIENTIF

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

Diagnosis	totaln	Positiven	Equivocaln	Negativen
Systemic lupus erythematosus	120	57	4	59
Primary Sjögren's syndrome	60	30	3	27
Idiopathic inflammatory myopathies (IIM) [total]	94	34	1	59
IIM PM	40	7	0	33
IIM DM	13	3	1	9
IIM Myositis/CTD-overlap	18	3	0	15
IIM Overlap / MCTD	9	9	0	0
IIM Jo-1 Positive	10	9	0	1
IIM Sporadic inclusion body myositis (sIBM)	4	3	0	1
Systemic sclerosis (SSc) [total]	91	19	5	67
SSc, diffuse	56	11	3	42
SSc, limited	27	8	2	17
SSc, various*	8	0	0	8
Celiac disease	13	0	0	13
Crohn's disease	12	0	0	12
CTD overlap Non-MCTD	10	0	1	9
Graves' disease	12	0	0	12
Primary antiphospholipid syndrome	12	1	0	11
Primary Biliary Cholangitis	25	1	2	22
Primary Sclerosing Cholangitis	24	0	0	24
Type 1 Diabetes	12	0	0	12
Diagnosis	totaln	Positiven	Equivocaln	Negativen
Ulcerative colitis	35	0	1	34
Lymphoma	21	0	0	21
Leukemia	20	0	0	20
Varied Cancer	9	0	0	9
Mixed connective tissue disease	10	2	0	8
Rheumatoid arthritis	35	2	0	33
Bacterial infections [total]	17	1	0	16
Bacterial infections (Mycoplasma)	3	0	0	3
Bacterial infections (Borrelia)	7	0	0	7
Bacterial infections (var. Staphylococcus)	3	1	0	2
Bacterial infections (various)	4	0	0	4
Viral infections [total]	80	4	1	75
Viral infections (Dengue)	4	0	0	4
Viral infections (EBV)	3	0	0	3
Viral infections (HBV)	11	1	0	10
Viral infections (HCV)	31	2	1	28
Viral infections (HIV)	24	1	0	23
Viral infections (various)	7	0	0	7
Hashimoto's disease	10	1	0	9
Vasculitis [total]	33	0	0	33
Granulomatosis with Polyangiitis	10	0	0	10
Eosinophilic granulomatosis withpolyangiitis	4	0	0	4
Microscopic polyangiitis	13	0	0	13
Polyarteritis nodosa	3	0	0	3
Giant cell arteritis	3	0	0	3
Target diseases	365	140	13	212
Control disease	390	12	5	373
Total	755	152	18	585

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*"SSc various" group is composed of the patients having SSc, classified according to the ACR/EULAR van den Hoogen 2013 criterion. The patients were not further sub-grouped.

{28}------------------------------------------------

EliA Ro60:

In total, 713 clinically and ethnically defined serum samples, including those of US origin, were used to determine sensitivity and specificity of the assay. Samples with a diagnosis of systemic lupus erythematosus (SLE) and Sjögren's syndrome (SS) represent the diagnostic group (target diseases: SLE n=120 + Sjögren's syndrome n=60). Samples with various autoimmune and infectious disease diagnoses represent the disease control group (n=533). The results are summarized in the tables below.

Results of clinically defined samples. SLE samples represented the "target disease" Sjögren's syndrome samples were excluded, other autoimmune diseases and infectious diseases were used as "control disease", but without control group SSc since these patients may have Ro antibodies. Equivocal results were either judged as negative or positive.

Equivocal = negative

Result	SLE	Controls	Total
Positive	58	6	64
Negative	62	436	498
Total	120	442	562

	Value	95% CI
Sensitivity	48.3%	39.1 - 57.6
Specificity	98.6%	97.1 - 99.5

Equivocal = positive
Result	SLE	Controls	Total
Positive	61	7	68
Negative	59	435	494
Total	120	442	562

Larring onl - mostition

	Value	95% CI
Sensitivity	50.8%	41.6 - 60.1
Specificity	98.4%	96.8 - 99.4

Results of clinically defined samples. Sjögren's syndrome samples represented the "target disease", SLE samples were excluded, other autoimmune diseases and infectious diseases were used as "control disease", but without control group SSc since these patients may have Ro antibodies. Equivocal results were either judged as negative or positive.

Equivocal = negative
Result	SS	Controls	Total
Positive	41	6	47
Negative	19	436	455
Total	60	442	502

	Value	95% CI
Sensitivity	68.3%	55 - 79.7
Specificity	98.6%	97.1 - 99.5

Equivocal = positive
Result	ટેટ	Controls	Total
Positive	43		રે રેણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામમાં મુખ્યત્વે ખેત-ઉપયોગ વિદ્યારત તેમ જ દૂધની ડેર
Negative	17	435	452
Total	60	442	502

	Value	95% CI
Sensitivity	71.7%	58.6 - 82.5
Specificity	98.4%	96.8 - 99.4

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I E N

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

Diagnosis	total n	positive n	equivocal n	negative n
Systemic lupus erythematosus (SLE)	120	58	3	59
Sjögren's syndrome (SS)	60	41	2	17
Idiopathic inflammatory myopathies (IIM)	10	0	0	10
Systemic sclerosis (SSc) [total]	91	21	0	70
SSc, diffuse	56	11	0	45
SSc, limited	27	10	0	17
SSc, various*	8	0	0	8
Celiac disease	13	0	0	13
Crohn's disease	12	0	0	12
CTD overlap Non-MCTD	10	1	0	9
Graves' disease	12	0	0	12
Primary antiphospholipid syndrome	12	1	0	11
Primary Biliary Cholangitis	25	0	0	25
Primary Sclerosing Cholangitis	24	0	0	24
Type 1 Diabetes	12	0	0	12
Ulcerative colitis	35	1	0	34
Lymphoma	21	0	0	21
Leukemia	20	0	0	20
Varied Cancer	9	0	0	9
Mixed connective tissue disease	10	0	0	10
Rheumatoid arthritis	77	1	1	75
Bacterial infections [total]	17	0	0	17
Bacterial infections (Mycoplasma)	3	0	0	3
Bacterial infections (Borrelia)	7	0	0	7
Bacterial infections (var. Staphylococcus)	3	0	0	3
Bacterial infections (various)	4	0	0	4
Viral infections [total]	80	1	0	79
Viral infections (Dengue)	4	0	0	4
Viral infections (EBV)	3	0	0	3
Viral infections (HBV)	11	0	0	11
Viral infections (HCV)	31	1	0	30
Viral infections (HIV)	24	0	0	24
Viral infections (various)	7	0	0	7
Diagnosis	totaln	positiven	equivocaln	negativen
Hashimoto's disease	10	1	0	9
Vasculitis [total]	33	0	0	33
Granulomatosis with Polyangiitis	10	0	0	10
Eosinophilic granulomatosis withpolyangiitis	4	0	0	4
Microscopic polyangiitis	13	0	0	13
Polyarteritis nodosa	3	0	0	3
Giant cell arteritis	3	0	0	3
Target diseases	180	99	5	76
Control disease	533	27	1	505
Total	713	126	6	581

The table below shows the results for each clinical subgroup:

Confidential Page 24 (26)

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*"SSc various" group is composed of the patients having SSc, classified according to the ACR/EULAR van den Hoogen 2013 criterion. The patients were not further sub-grouped.

d) Other Clinical Supportive Data:

Not applicable.

Clinical Cut-Off: Same as assay cut-off.

Expected Values/Reference Range:

Antibody prevalence in autoimmune patients varies widely depending on disease area. The proportion of sera from a normal population found positive for the antinuclear antibodies covered by the EliA Ro52 and EliA Ro60 test is below 1%. Expected values may vary depending on the population tested.

The frequency distribution for antinuclear antibodies was investigated in a group of apparently healthy subjects equally distributed by age and gender, using sera from Caucasian, African American, Hispanic and Asian population obtained from a blood bank.

The results are given in the table below:

Test	n	MedianEliA U/mL	95th percentileEliA U/mL	99th percentileEliA U/mL
Elia Ro52	638	1.0	1.8	2.6
Elia Ro60	638	0.5	0.9	7.4

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Proposed Labeling

The labeling is drafted in accordance with the requirements of 21 CFR Part 809.10.

Conclusion

All available data support that both immunoassays, the new devices EliA Ro52 and EliA Ro60 and their proposed predicate devices QUANTA Flash Ro52 and QUANTA Flash Ro60 perform substantially equivalent.

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

Regulation Number and Section

§ 866.5100 Antinuclear antibody immunological test system.

(a)
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).(b)
Classification. Class II (performance standards).