Who is the manufacturer of ELIA SCL-70S IMMUNOASSAY?

Phadia GmbH filed the 510(k) submission for ELIA SCL-70S IMMUNOASSAY (K140493).

What is the FDA product code for ELIA SCL-70S IMMUNOASSAY?

LJM. It is classified under 21 CFR 866.5100 as a Class 2 device in the Immunology panel.

What is the regulatory pathway for ELIA SCL-70S IMMUNOASSAY?

510(k) clearance (submission type: Traditional).

What are the predicate devices for ELIA SCL-70S IMMUNOASSAY?

QuantaLite™ Scl-70 ELISA, INOVA (K924988).

Who can help prepare an FDA 510(k) submission like ELIA SCL-70S IMMUNOASSAY?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA 510(k) submissions. See https://innolitics.com/services/510ks/ or contact https://innolitics.com/contact.

ELIA SCL-70S IMMUNOASSAY

K140493 · Phadia GmbH · LJM · Oct 30, 2014 · Immunology

Device Facts

Record ID	K140493
Device Name	ELIA SCL-70S IMMUNOASSAY
Applicant	Phadia GmbH
Product Code	LJM · Immunology
Decision Date	Oct 30, 2014
Decision	SESE
Submission Type	Traditional
Regulation	21 CFR 866.5100
Device Class	Class 2

Intended Use

EliA Scl-70S is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70S uses the EliA IgG method on the instrument Phadia 100. EliA Scl-70S is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70S uses the EliA IgG method on the instrument Phadia 250.

Device Story

EliA Scl-70S is an in vitro diagnostic immunoassay for detecting IgG antibodies to Scl-70. The device uses microwells coated with human recombinant Scl-70 protein. Patient serum or plasma samples are incubated in the wells; if anti-Scl-70 antibodies are present, they bind to the antigen. After washing, enzyme-labeled anti-human IgG (beta-galactosidase) is added to form an antibody-conjugate complex. Following a second wash, a development solution is added; the resulting fluorescence is measured by the Phadia 100 or Phadia 250 automated instrument. The instrument compares the fluorescent signal against a stored calibration curve to quantify the antibody concentration. Results are reported in EliA U/mL. The device is intended for use in clinical laboratories by trained personnel. By identifying Scl-70 antibodies, the test assists clinicians in diagnosing diffuse scleroderma, supporting clinical decision-making when combined with other diagnostic criteria.

Clinical Evidence

Clinical performance was evaluated using 336 clinically defined serum samples (101 Scleroderma, 235 controls including CREST, MCTD, SLE, SS, PM/DM, RA, cancer, and infections). Sensitivity was 30.7% (95% CI: 22.1%–40.8%) and specificity was 98.7% (95% CI: 96.0%–99.7%). Method comparison with the predicate (n=374) showed 95.9% total agreement (equivocal as negative). Analytical studies included precision (Total CV 3.33%–6.03%), linearity (0.6–297.1 EliA U/mL), and interference testing (no interference up to specified concentrations of bilirubin, hemoglobin, chyle, and rheumatoid factor).

Technological Characteristics

Solid-phase immunoassay using microwells coated with human recombinant Scl-70 protein. Detection via beta-galactosidase-labeled anti-human IgG conjugate and 4-methylumbelliferyl-beta-D-galactoside development solution. Fluorescence-based signal detection. Automated operation on Phadia 100/250 systems. Calibrated against WHO IRP 67/86. Reagents include PBS, BSA, detergent, and sodium azide preservatives. Controlled 37°C incubation.

Indications for Use

Indicated for the semi-quantitative measurement of IgG antibodies to Scl-70 in human serum and plasma to aid in the clinical diagnosis of diffuse scleroderma in conjunction with other clinical and laboratory findings.

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

QuantaLite™ Scl-70 ELISA, INOVA (K924988)

Reference Devices

EliA Ro (K082759)
EliA ANA Positive Control (K063775)
EliA IgG/IgM/IgA Negative Control (K072393, K091845)

Related Devices

K983423 — THE APTUS (AUTOMATED) APPLICATION OF THE SCL-70 ELISA TEST SYSTEM. AN ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE · Zeus Scientific, Inc. · Nov 18, 1998
K990797 — AUTOSTAT II ANTI-SCL-70 ELISA · Cogent Diagnotics , Ltd. · Apr 28, 1999
K970239 — IS-ANTI-SCL-70 TEST SYSTEM (720-280) · Diamedix Corp. · Apr 8, 1997
K113143 — IMMULISA SCI-70 ANTIBODY ELISA · Immco Diagnostics, Inc. · Dec 2, 2011
K955602 — RELSIA SCL-70 ANTIBODY TEST SYSTEM · Immuno Concepts, Inc. · Apr 19, 1996

Submission Summary (Full Text)

{0}------------------------------------------------ Image /page/0/Picture/1 description: The image is a black and white logo for the U.S. Department of Health & Human Services. The logo features a stylized caduceus symbol, which is often associated with healthcare. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the symbol. Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002 October 30, 2014 PHADIA US, INC. C/O MR. MARTIN ROBERT MANN SENIOR REGULATORY AFFAIRS MANAGER 4169 COMMERCIAL AVENUE PORTAGE, MI 49002 Re: k140493 Trade/Device Name: EliA™ Scl-708 Immunoassay Regulation Number: 21 CFR §866.5100 Regulation Name: Antinuclear Antibody Immunological Test System Regulatory Class: Class II Product Code: LJM Dated: September 24, 2014 Received: September 26, 2014 Dear Mr. Mann: 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. 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 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); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. {1}------------------------------------------------ If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance. You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Sincerely vours. # Maria M. Chan -S Maria M. Chan, Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health Enclosure {2}------------------------------------------------ # Indications for Use 510(k) Number (if known) K140493 Device Name EliA™ Scl-70s ### Indications for Use (Describe) EliA Scl-70s is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70s uses the EliA IgG method on the instrument Phadia 100. EliA Scl-70s is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70s uses the EliA IgG method on the instrument Phadia 250. | 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. This section applies only to requirements of the Paperwork Reduction Act of 1995. # *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: > Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {3}------------------------------------------------ ### 510k Summary long EliA Scl-70S D.1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION SUMMARY - A. 510(k) Number: k140493 - B. Purpose for Submission: New devices - C. Measurand: lgG Antibodies to Scl-70 - D. Type of Test: Semi-quantitative immunofluorescence assay - E. Applicant: Phadia US Inc. - F. Proprietary and Established Names: EliA™ Scl-708 Well - G. Regulatory Information: 1. Regulation section: 21 CFR §866.5100, Antinuclear antibody immunological test system - 2. Classification: Class II (Assay) - 3. Product code: LJM Antinuclear Antibody (Enzyme-Labeled), Antigen, Controls| - 4. Panel: Immunology {4}------------------------------------------------ #### H. Intended Use: ### 1. Intended use(s): EliA Scl-70° is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70 uses the EliA IgG method on the instrument Phadia 100. EliA Scl-70° is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70 uses the EliA IgG method on the instrument Phadia 250. - 2. Indication(s) for use: Same as intended use - 3. Special conditions for use statement(s): For prescription use only - 4. Special instrument requirements: Phadia® 100, Phadia® 250 {5}------------------------------------------------ ### l. Device Description: - EliA Scl-70 Wells (14-5637-01) are coated with a human recombinant Scl-70 protein -4 carriers, (12 wells each), ready to use The method-specific reagents on Phadia 250 are identical; they are only filled in different containers. Needed reagents for Phadia 100: - EliA Sample Diluent, method-specific, PBS containing BSA, detergent and 0.095% sodium azide . - ready to use: (83-1003-01) - EliA IgG Conjugate, ß-galactosidase labeled anti-IgG (mouse monoclonal antibodies) in PBS containing BSA, detergent and 0.06% sodium azide - ready to use; (83-1002-01, 83-1005-01) - EliA ANA Positive Control 100: 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; (83-1038-01) - EliA IgG/IgM/IgA Negative Control 100: containing normal human serum from healthy donors -. 6 single-use vials, 0.3 mL each, ready to use; (83-1042-01) - EliA IgG Calibrators, human IgG in PBS (0, 4, 10, 20, 100, 600 µg/L) 6 single-use vials, 0.3 mL . each, ready to use; (83-1000-01) - EliA IgG Curve Control, human IgG (20 µg/L) in PBS, 6 single-use vials, 0.3 mL each, ready to . use; (83-1001-01) - EliA IgG Calibrator Wells are coated with mouse monoclonal antibodies 4 carriers. (12 wells . each), ready to use; (14-5509-01) Needed reagents for Phadia 250: - EliA Sample Diluent, method-specific, PBS containing BSA, detergent and 0.095% sodium azide . - ready to use; (83-1023-01) - EliA IgG Conjugate, ß-galactosidase labeled anti-IgG (mouse monoclonal antibodies) in PBS containing BSA, detergent and 0.06% sodium azide - ready to use; (83-1017-01, 83-1018-01) - . EliA ANA Positive Control 250: 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; (83-1033-01) - EliA IgG/IgM/IgA Negative Control 250: containing normal human serum from healthy donors -. 6 single-use vials, 0.3 mL each, ready to use; (83-1037-01) - EliA IgG Calibrator Strips, human IgG in PBS (0, 4, 10, 20, 100, 600 µg/L) 6 single-use vials, . 0.3 mL each, ready to use; (83-1015-01) - EliA IgG Curve Control Strips, human IgG (20 µg/L) in PBS, 6 single-use vials, 0.3 mL each, . ready to use; (83-1016-01) - EliA IgG Calibrator Wells are coated with mouse monoclonal antibodies 4 carriers, (12 wells each), ready to use; (14-5509-01) The Phadia EliA immunodiagnostic system is automated system for immunodiagnostic testing. The EliA reagents are available as modular packages, each purchased separately. All packages except the positive and neqative controls are required to carry out an EliA Scl-70° test. ### J. Substantial Equivalence Information: - 1. Predicate device name(s) and 510(k) number(s): QuantaLite™ Scl-70 ELISA, INOVÀ K924988 {6}------------------------------------------------ # 2. Comparison with predicate: # Similarities to predicate device | Feature | Predicate Device | New Device | |-------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | | INOVA QuantaLite Scl-70 ELISA | EliA Scl-70S | | Intended Use | QUANTALite Scl-70 is an en- zyme-linked immunosorbent as- say (ELISA) for the semi- quantitative detection of Scl-70 antibodies in human serum. The presence of Scl-70 antibodies can be used in conjunction with clinical findings and other labora- tory tests to aid in the diagnosis of scleroderma. | EliA Scl-70S is intended for the in vitro semi-quantitative measure- ment of IgG antibodies directed to Scl-70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (diffuse form) in conjunction with other laboratory and clinical findings. EliA Scl-70S uses the EliA IgG method on the instruments Phadia 100 or Pha- dia 250. | | Internal Controls | Low Positive, High Positive and Negative Control included in the kit | Positive and negative Control provided with the EliA ANA Posi- tive Control 100 / 250 and EliA IgG/IgM/IgA Negative Control 100 / 250, respectively | | Assay Type | ELISA | | | Type of test | Semi-quantitative | | | Solid Phase | Microwells | | ### Differences to predicate device | Differences to predicate device | | | |-----------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------| | Feature | Predicate Device INOVA QuantaLite Scl-70 ELISA | New Device EliA Scl-70S | | Antigen used | purified Scl-70 antigen | Human recombinant Scl-70 pro- tein | | Instrumentation | ELISA-Reader needed | Phadia 100, Phadia 250 are fully automated immunoassay ana- lyzers | | Reaction tempera- ture | Room temperature, 20-26°C | 37°C controlled | | Incubation times | High positive, low positive and negative Controls, diluted patient samples: 30 min. Conjugate: 30 min. Substrate: 30 min (in dark). | Diluted patient samples: 30 min. Conjugate: 28 min. Development Solution: 39 min. | | Detection antibody (conjugate) | IgG conjugate: anti-human IgG horse radish peroxidase (goat) | IgG conjugate: anti-human IgG β-Galactosidase (mouse mono- clonal antibodies) | | Signal | Optical density (at 450nm) | Fluorescence | | Calibration | 1-point Calibration | Total IgG Calibration | | Calibrators | n.a. | 6 vials of human IgG at concen- trations of 0 - 4 - 10 - 20 - 100 - 600 µg/l | | Calibration curve | n.a. | Option to store curve for up to 28 days and run curve controls in each assay for calibration | | Concept | All reagents in a single kit | Modular reagents concept (test-method specific and general | {7}------------------------------------------------ | Feature | Predicate Device INOVA QuantaLite Scl-70 ELISA | New Device EliA Scl-70S reagents) | |-----------------|-----------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Sample Dilution | 1:101 | 1:100 | | Reported Unit | Units | EliA U/ml | | Cut-off | < 20 Units negative 20-39 Units weak positive 40-80 Units moderate positive > 80 Units strong positive | < 7.0 EliA U/ml negative 7.0-10.0 EliA U/ml equivocal > 10.0 EliA U/ml positive | | Substrate | TMB Chromogen | Development Solution 0.01 % 4-Methylumbelliferyl-β-D- galactoside & <0.0010% pre- servative (mixture of 5-chloro-2- methyl-2H-isothiazol-3-one [EC no. 247-500-7] and 2-methyl-2H- isothiazol-3-one [EC no. 220- 239-6] (3:1)) | {8}------------------------------------------------ ### K. Standard/Guidance Document Referenced (if applicable): CLSI EP17-A, Protocols for Determination of Limits of Detection and Limits of Quantification, Approved Guideline. ### L. Test Principle: The EliA Scl-708 Wells are coated with human recombinant Scl-70 protein. If present in the patient's specimen, antibodies to Scl-70 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. ### M. Performance Characteristics (if/when applicable): 1. Analytical performance: ### Precision/Reproducibility: a. To determine the precision of the assays on Phadia 250 instruments, the variability was assessed on 3 samples. One batch was used to determine the precision of the assays on Phadia 100. Three batches were used to determine the precision of the assays on Phadia 250 (equal to 252 replicate determinations per sample). The results are summarized in the tables below: To determine the precision of the assays, the variability was assessed in a study with a total of 21 runs (3 instruments x 7 runs each) over 7 days performed on samples with 84 replicate determinations per sample with a calibration curve in each run. | Mean value EliA U/ml | Intra-Run CV% | Inter-Run CV% | Total %CV | |-------------------------|------------------|------------------|--------------| | 8.8 | 2.89 | 2.06 | 3.55 | | 30.2 | 3.21 | 2.41 | 4.01 | | 193.0 | 4.90 | 3.51 | 6.03 | # EliA Scl-708 on Phadia 100 (n=84) To determine the precision of the assay, the variability was assessed in a study with a total of 21 runs (3 instruments x 7 runs each) over 7 days performed on samples with 252 replicate determinations per sample with a calibration curve in each run. | Mean value EliA U/ml | Intra Run CV% | Inter Run CV% | Total %CV | Lot to Lot CV% | |-------------------------|------------------|------------------|--------------|-------------------| | 7.5 | 3.08 | 1.25 | 3.33 | 1.07 | | 28.2 | 2.71 | 2.26 | 3.53 | 2.01 | | 200.1 | 4.46 | 3.05 | 5.40 | 5.27 | ### EliA Scl-70 on Phadia 250 (n = 252) {9}------------------------------------------------ #### Linearity/assay reportable range: b. Four patient serum samples were diluted in sample diluent and tested with one batch of EliA Scl-70° with one set of system reagents on Phadia 250. The ratios of observed/expected values and least square linear regressions were calculated. The results are summarized below: EliA Scl-708 on Phadia 100 | Dilution range (EliA U/ml) | Slope | Intercept | R2 | |----------------------------|-------|-----------|------| | 172.3 - 1.8 | 1.112 | -0.902 | 1.00 | | 185.5 - 2.1 | 0.995 | 0.151 | 1.00 | | 192.0 - 2.1 | 1.062 | 0.300 | 1.00 | | 28.1 - 0.1 | 0.960 | -0.142 | 1.00 | | 297.1 - 2.9 | 1.101 | -1.134 | 1.00 | EliA Scl-708 on Phadia 250 | Dilution range (EliA U/ml) | Slope | Intercept | R2 | |-------------------------------|-------|-----------|------| | 207.7 - 1.9 | 0.961 | -0.399 | 1.00 | | 159.9 - 1.4 | 1.088 | 0.655 | 1.00 | | 193.6 - 2.1 | 1.062 | 0.047 | 1.00 | | 24.3 - 0.2 | 1.041 | 0.040 | 1.00 | | 298.9 - 3.1 | 1.088 | 0.653 | 1.00 | The claimed linear range for EliA Scl-70° is 0.6 - 297.1 EliA U/ml (0.6 is the limit of detection). # HookEffect/OvertheRangeResults: Hook effect was investigated by using a serum sample above the highest calibration point. A high positive sample was diluted and the dilutions were measured in four replicates and compared to Cal-600 (600 µg/L). No hook effect was observed when analyzing a high positive sample that had a concentration up to 14 times above the upper limit of the technical measuring range. Results above the upper limit of the measuring range are reported as "above". No recommendations are made for dilution of samples outside measuring range in the Package Insert. Traceability, Stability, Expected values (controls, calibrators, or methods): C. 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 as bar code on the EliA well, the results are automatically converted to EliA U/mL for EliA Scl-70°. # Stability: The EliA IgG system reagents EliA ANA Positive Control and the EliA IgG/IgM/IgA Negative Control are already FDA cleared under k063775, k072393, and k091845, respectively. {10}------------------------------------------------ ### Shelf life: Accelerated study was done to determine the shelf life EliA Scl-70 Wells and was determined to be 18 months at 2-8°C. The real time stability test confirms the determined shelf life. ### On-board stability: The on-board stability EliA Scl-70 carriers was tested over 4 weeks using 3 positive and 2 negative samples only on the Phadia 250 instrument since for Phadia 100 the reagents are stored off the instrument and are only loaded as needed for an assay. The on-board stability for the Phadia 250 instrument was determined to be 28 days at 2-8°C. ### Open Stability: Accelerated study was done to determine the shelf life EliA Scl-70° Wells after first opening of the foil bag containing the EliA Scl-70° Wells. It was determined to be 9 months at 2-8°C. The real time stability test confirms the determined shelf life. #### d. Detection limit: The limit of blank (LoB) and limit of detection (LoD) studies were done on both Phadia 100 and Phadia 250. Six blood donors with low/negative antibody concentration were measured in twelve replicates in each of six runs on six different days (12 replicates x 6 runs x 6 days = 432 replicates per sample). The six runs for each system, Phadia 100 and Phadia 250, were spread over two instruments. The results are summarized in the tables below: | EliA Scl-70S (EliA U/mL) | LoB | LoD | |--------------------------|------|------| | Phadia 100 | 0.25 | 0.36 | | Phadia 250 | 0.31 | 0.52 | It was decided to use a single LoD of 0.6 U/mL for both instruments. ### Analytical specificity: e. ### Endogenous Interference: A study was run to investigate whether high concentrations of potentially interfering substances in serum, like bilirubin, hemoglobin, chyle and rheumatoid factor adversely affect the results of the new device. Five serum samples were prediluted in EliA Sample Diluent and spiked with the different interfering substances or their respective blank solutions, and analyzed in triplicates. The runs were repeated twice. One batch of EliA antigen wells and one batch of system reagents were used throughout the studies. Two negative, two samples with concentrations around the cut-off and one positive sample were tested. The ratio of blank/spiked sample was 0.94 – 1.09. No interference was observed up to the concentrations listed in the table below: | Potential Interfering Compound | Concentration in undiluted sample | |--------------------------------|-----------------------------------| | Bilirubin F | 19.2 mg/dl | | Bilirubin C | 20.1 mg/dl | | Hemoglobin | 496 mg/dl | | Chyle (ClinOleic) | 1% | | Rheumatoid factor | 500 IU/ml | # Carry-over: Carry over is a technical phenomenon that is due to the instrumentation. The instrumentation is identical for all EliA tests (Phadia instruments). Also the basic circumstances of the test are identical between EliA Ro and the newly submitted EliA test (same source of test material, same reagents, same magnitude of specific antibody concentration etc.). Thus, results obtained with one EliA test can be transferred to any other EliA serum test. {11}------------------------------------------------ A study was carried out on a Phadia 250 instrument using the test EliA Ro, cleared under k082759. A serum sample was diluted 1:2 and 1:20 using instrument dilution and manual dilution. A lower dilution factor than the default one (1:100) was chosen to challenge the system. Only a few RUs difference compared to the reference pipetting could be seen, which is too low to be expressed in EliA U/ml. The observed carry over effect is therefore negligible without any influence to assay results. #### f. Assay cut-off: A study was done on 400 apparently healthy blood donor samples from Caucasian individuals equally distributed by sex and age in order to evaluate expected values in the normal population and to confirm the defined cut-off. The samples were measured on the Phadia 250 instrument for EliA Scl-70°. The 99th percentile of the 400 samples were calculated and taken into account for setting of the cut-off. The following values were selected for the cut-off: | EliA Scl-708 | | |------------------|-----------| | <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: #### Method comparison with predicate device: a. A total of 390 serum samples were collected from patients with diagnosis of Scleroderma (SSc. n = 101), limited cutaneous form of systemic sclerosis (CREST, n = 33), Mixed connective tissue disease (MCTD, n = 37), Systemic lupus erythematodes (SLE, n = 34), Sjögren's syndrome (SS, n = 26), Poly/Dermatomyositis (PM/DM, n = 5), Rheumatoid arthritis (RA, n = 30), various cancers (n = 20), various bacterial infections (n = 24), various viral infections (n = 26), technical samples positive or equivocal for EliA Scl-70° (n = 54). Samples were analyzed with the EliA Scl-70° and INOVA QuantaLite™ Scl-70 ELISA assays. The test was run in single determination and values outside the linear measuring range (n=16) were excluded from statistical analyses for the technical agreement. The results are summarized in the tables below: | QuantaLite Scl-70 ELISA (Units) | | | | | |-----------------------------------------------------------------|--------------|--------------|-------|-----| | | Positive >20 | Negative <20 | Total | | | EliA Scl-70S (EliA U/mL) | Positive >10 | 51 | 9 | 60 | | | Negative ≤10 | 6 | 308 | 314 | | | Total | 57 | 317 | 374 | | Positive percent agreement: 89.5% (95% CI: 77.8 – 95.6%) | | | | | | Negative percent agreement: 97.2% (95% CI: 94.5 – 98.6%) | | | | | | Total percent agreement: 95.9% (95% CI: 93.3 – 97.7%) | | | | | Equivocal results evaluated as negative: {12}------------------------------------------------ Equivocal results evaluated as positive: | | | QuantaLite Scl-70 ELISA (Units) | | | |-------------------------------------------------------------|--------------|---------------------------------|--------------|-------| | | | Positive >20 | Negative <20 | Total | | EliA Scl-70S (EliA U/mL) | Positive >10 | 53 | 21 | 74 | | | Negative ≤10 | 4 | 296 | 300 | | | Total | 57 | 317 | 374 | | Positive percent agreement: 93.0% (95% CI: 82.2 – 97.8%) | | | | | Negative percent agreement: 93.4% (95% Cl: 89.9 - 95.8%) Total percent agreement: 93.3% (95% CI: 90.2 - 95.5%) #### b. Matrix comparison: Serum, lithium heparin plasma, and EDTA plasma were collected from the same patients (n = 50) to demonstrate that the plasma results do not deviate from the corresponding serum results and are within the pre-defined specifications. Samples were spiked with a serum sample of high antibody titer to cover the measuring range. Samples were tested in duplicates. Passing & Bablok regression plots were generated using the first replicate only and by plotting the concentration observed from the control tube (serum) versus the concentration for each test collection tube. The corresponding slopes of regression and coefficient determination are summarized in the tables below: | | Range tested (EliA U/mL) | Slope (95% CI) | Intercept (95% CI) | R2 | |----------------------------|-----------------------------|------------------------|---------------------------|------| | Serum vs heparin plasma | 0.6 — 278.9 | 1.05 (1.03 to 1.07) | -0.08 (-0.24 to +0.17) | 1.00 | | Serum vs EDTA plasma | 0.6 — 264.2 | 1.00 (0.99 to 1.02) | -0.05 (-0.29 to +0.21) | 1.00 | #### C. Instrument comparison Performance of EliA Scl-70° was evaluated on the Phadia 250 instruments using 24 positive, 8 equivocal and 4 negative samples were analyzed in six runs in single replicates on three Phadia 250 instruments, with 2 runs on each instrument. The regression analysis results are summarized as follows: | | Intercept | Slope | |----------|--------------|-------------| | Estimate | -0.07 | 0.96 | | 95% Cl | -0.52 – 0.14 | 0.94 – 0.99 | # 3. Clinical studies: Clinical Sensitivity and specificity: a. 336 clinically defined sera from patients with scleroderma or other connective tissue diseases, bacterial or viral infections, cancer or rheumatoid arthritis were tested on EliA Scl-70° as laid out in the respective Directions for Use {13}------------------------------------------------ Test results of EliA Scl-70 with 336 clinically defined samples. The diagnostic group of Scleroderma diagnosed patients (SSc) is the positive target group. Equivocal results evaluated as negative: | | Diagnostic Group - SSc | | | |-----------------------------|------------------------|---------------|------------| | | + / SSc | - / Controls* | total | | Positive test >10 EliA U/mL | 31 | 3 | 34 | | Negative test ≤10 EliA U/mL | 70 | 232 | 302 | | Total | 101 | 235 | 336 | Sensitivity: 30.7% (95% CI: 22.1% - 40.8%) Specificity: 98.7% (95% CI: 96.0% - 99.7%) The table below shows the positive results for each clinical subgroup. Equivocal results evaluated as negative: | Disease | Total number of samples | Number & percentage of positive samples QuantaLite Scl-70 | | Elia Scl-70s | | |------------|----------------------------|--------------------------------------------------------------|-----|--------------|-----| | SSc | 101 | 29 | 29% | 31 | 31% | | CREST | 33 | 2 | 6% | 2 | 6% | | MCTD | 37 | 0 | 0% | 0 | 0% | | SLE | 34 | 1 | 3% | 0 | 0% | | SS | 26 | 1 | 4% | 1 | 4% | | PM/DM | 5 | 0 | 0% | 0 | 0% | | RA | 30 | 0 | 0% | 0 | 0% | | Cancer | 20 | 0 | 0% | 0 | 0% | | Infections | 50 | 2 | 4% | 0 | 0% | SSc: Scleroderma, CREST: limited cutaneous form of systemic sclerosis, MCTD: Mixed connective tissue disease, SLE: Systemic Lupus Erythematosus, SS: Sjögren's syndrome- systemic sclerosis, PM/DM: Poly-/Dermatomyositis, RA: Rheumatoid arthritis, Cancer: various cancers, Infections: various bacterial and viral infections. Other clinical supportive data (when a. is not applicable): b. Not applicable - 4. Clinical cut-off: Same as assay cut-off - 5. 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 Scl-70 antibodies covered by the EliA Scl-70 test is below 1 %. Expected values may vary depending on the population tested.