EliA CENP Immunoassay, EliA U1RNP Immunoassay, EliA RNP70 Immunoassay
Device Facts
| Record ID | K182353 |
|---|---|
| Device Name | EliA CENP Immunoassay, EliA U1RNP Immunoassay, EliA RNP70 Immunoassay |
| Applicant | Phadia AB |
| Product Code | LJM · Immunology |
| Decision Date | Nov 27, 2018 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.5100 |
| Device Class | Class 2 |
Intended Use
EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA CENP uses the EliA IgG method on the instrument Phadia 2500/5000. EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument Phadia 2500/5000. EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 uses the EliA IgG method on the instrument Phadia 2500/5000.
Device Story
EliA CENP, U1RNP, and RNP70 immunoassays are semi-quantitative in vitro diagnostic tests for IgG antibodies in human serum/plasma. The device uses polystyrene wells coated with specific recombinant antigens (CENP, U1RNP, or RNP70). Patient samples are incubated in these wells; specific antibodies bind to the antigen. After washing, enzyme-labeled anti-IgG conjugate is added, followed by a development solution. The Phadia 2500/5000 instrument measures the resulting fluorescence, which is proportional to the antibody concentration. The system is operated by laboratory personnel in a clinical setting. Results are compared to calibrators to determine antibody levels. The output aids clinicians in diagnosing connective tissue diseases. The device is not for point-of-care use.
Clinical Evidence
Bench testing only. Precision/reproducibility studies (21 runs, 84 replicates per sample) showed total imprecision (%CV) ranging from 6.0% to 24.4%. Linearity was confirmed with R2 values of 0.99-1.00. Method comparison studies (n > 100 samples) between Phadia 250 and Phadia 2500/5000 met acceptance criteria for slope (0.9-1.1) and intercept. LoD/LoQ were established per CLSI EP17-A. No new clinical diagnostic sensitivity/specificity studies were required as these were previously reviewed for the predicate assays.
Technological Characteristics
Solid-phase fluoroimmunoassay; recombinant protein-coated wells; β-Galactosidase labeled anti-IgG conjugate; 4-Methylumbelliferyl-β-D-galactoside development solution. Automated platform (Phadia 2500/5000) with disposable pipette tips. Incubation at 37°C. Connectivity via IDM software. Sterilization not applicable (reagents).
Indications for Use
Indicated for patients suspected of having scleroderma (CREST Syndrome), mixed connective tissue disease (MCTD), or systemic lupus erythematosus (SLE). Used as an aid in clinical diagnosis in conjunction with other laboratory and clinical findings. Not for point-of-care use.
Regulatory Classification
Identification
An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).
Predicate Devices
- EliA CENP on Phadia 250 (K082759)
- EliA U1RNP on Phadia 250 (K082759)
- EliA RNP70 on Phadia 250 (K083117)
Related Devices
- K083117 — ELIA RNP70 IMMUNOASSAY, ELIA SCI-70 IMMUNOASSAY AND ELIA JO-1 IMMUNOASSAY, MODELS 14-5511-01, 14-5506-01 AND 14-5507-01 · Phadia US, Inc. · May 28, 2009
- K072149 — ELIA SYMPHONY IMMUNOASSAY, ANA CONTROL, MODEL 14-55508-01,83-1004-01 · Phadia US, Inc. · Jan 15, 2008
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
A. 510(k) Number:
K182353
B. Purpose for Submission:
Addition of previously cleared assays on a new instrument platform (Phadia 2500/5000)
C. Measurand:
IgG autoantibodies specific to Centromere protein (CENP), U1 ribonucleoprotein (U1RNP), and ribonucleoprotein 70kDa (RNP70)
D. Type of Test:
Automated semi-quantitative solid-phase fluoroimmunoassays
E. Applicant:
Phadia AB
F. Proprietary and Established Names:
EliA CENP Immunoassay, EliA U1RNP Immunoassay, El RNP70 Immunoassay
G. Regulatory Information:
1. Regulatory Section:
21 CFR 866.5100, Antinuclear antibody immunological test system
2. Classification:
Class II
3. Product Codes:
LJM, Antinuclear Antibody (Enzyme-labeled), Antigen, Controls
LKO, Anti-RNP Antibody, Antigen and Control
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4. Panel:
Immunology (82)
H. Intended Use:
1. Intended uses:
EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA CENP uses the EliA IgG method on the instrument Phadia 2500/5000.
EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument Phadia 2500/5000.
EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 uses the EliA IgG method on the instrument Phadia 2500/5000.
2. Indication for uses:
Same as Intended Uses
3. Special conditions for use statement:
Prescription use only
4. Special instrument requirements:
For use on the Phadia 2500 and Phadia 5000 instruments
I. Device Description:
EliA uses a modular reagent system. The assay-specific, method-specific and general reagents are packaged and purchased as separate units. The reagents on Phadia 2500 and Phadia 5000 are identical; they are only filled in different containers.
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(1) EliA Assay-Specific reagent:
- EliA CENP Wells are coated with human recombinant centromere protein B in two carriers (12 wells each), ready to use;
- EliA U1RNP Wells are coated with human recombinant RNP (RNP70, A, C) proteins in four carriers (12 wells each), ready to use;
- EliA RNP70 Wells are coated with human recombinant RNP (70 kDa) protein in four carriers (12 wells each), ready to use;
(2) EliA Method-specific reagent:
- EliA IgG Calibrator Strips: Human IgG (0, 4, 10, 20, 100, 600 µg/L) in PBS containing BSA, detergent and 0.095% (w/v) sodium azide in five strips, six single-use vials per strip, 0.3 mL each, ready to use;
- EliA IgG Curve Control Strips: Human IgG (20 µg/L) in PBS containing BSA, detergent and 0.095% (w/v) sodium azide in five strips, six single-use vials per strip, 0.3 mL each, ready to use;
- EliA Sample Diluent: PBS containing BSA, detergent and 0.095% (w/v) sodium azide in six bottles, 48 mL each, ready to use; or six bottles, 400 mL each, ready to use;
- EliA IgG Conjugate 50 or 200: β-Galactosidase labeled anti-IgG (mouse monoclonal antibodies) in PBS containing BSA and 0.06% (w/v) sodium azide in 6 wedge-shaped bottles, 5 mL each, ready to use; or six wedge-shaped bottles, 19 mL each, ready to use;
- EliA IgG Calibrator Well: Coated with mouse monoclonal antibodies in four carriers (12 wells each), ready to use;
(3) EliA General reagents:
- Development Solution (0.01% 4-Methylumbelliferyl-β-D-galactoside, <0.0010% preservative), ready for use;
- Stop Solution (4% Sodium Carbonate), ready for use;
- Dilution Wells (high density polyethylene wells), 50 carriers, ready to use;
- Pipette Tips in Racks (polyethylene tips), 24 racks × 160 tips, ready for use;
- Washing Solution (information in separate Washing Solution package insert)
Phadia 2500 and Phadia 5000 are identical instruments except for sample throughput. The Phadia 2500 consists of one process module (two process lines), whereas Phadia 5000 consists of two process modules (2 x 2 process lines). Instrument operation is handled by onboard Instrument Software (ISW). Data output is administered by Information Data Manager (IDM). All steps of an assay are performed within a single process line. Thus, study protocols used for Phadia 2500 are also valid for Phadia 5000.
J. Substantial Equivalence Information:
1. Predicate device name:
EliA CENP on Phadia 250, EliA U1RNP on Phadia 250, EliA RNP70 on Phadia 250
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2. Predicate 510(k) number:
K082759 (EliA CENP and EliA U1RNP)
K083117 (EliA RNP70)
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | Predicate Device
Phadia 250 | Test Device
Phadia 2500/5000 |
| Intended Use/Indications for Use EliA CENP | EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (heparin, EDTA, citrate) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA CENP uses the EliA IgG method on the instrument ImmunoCAP 250. | EliA CENP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to CENP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of scleroderma (CREST Syndrome) in conjunction with other laboratory and clinical findings. EliA CENP uses the EliA IgG method on the instrument |
| Intended Use/Indications for Use EliA U1RNP | EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (heparin, EDTA, citrate) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument ImmunoCAP 250. | EliA U1RNP is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to U1RNP in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA U1RNP uses the EliA IgG method on the instrument |
| Intended Use/Indications for Use EliA RNP70 | EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 is to be used together with the EliA IgG method on the instrument ImmunoCAP 250. | EliA RNP70 is intended for the in vitro semi-quantitative measurement of IgG antibodies directed to RNP70 in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) in conjunction with other laboratory and clinical findings. EliA RNP70 uses the EliA IgG method on the instrument |
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| Similarities | | |
| --- | --- | --- |
| Item | Predicate Device
Phadia 250 | Test Device
Phadia 2500/5000 |
| Analytical technology | Immuno-fluorescence measurement | Same |
| Reagents | Common Phadia EliA assay components | Same
Introduction of new article numbers for Development Solution, Stop Solution and Washing Solution is only due to larger filling volumes which are required for the bigger instruments Phadia 2500/5000 |
| Result calculation software | Phadia Information Data Manager (IDM) | Same |
| Assay cutoff / results interpretation for EliA CENP | Negative: < 7 EliA U/mL
Equivocal: 7–10 EliA U/mL
Positive: > 10 EliA U/mL | Same |
| Assay cutoff / results interpretation for EliA U1RNP | Negative: < 5 EliA U/mL
Equivocal: 5–10 EliA U/mL
Positive: > 10 EliA U/mL | Same |
| Assay cutoff /results interpretation for EliA RNP70 | Negative: < 7 EliA U/mL
Equivocal: 7–10 EliA U/mL
Positive: > 10 EliA U/mL | Same |
| Calibrator | IRP 67/86 of Human Serum IgA, G, M from World Health Organization | Same |
| Sample volume | 90 μL (20 μL of non-diluted sample) | Same |
| Incubation temperature | 37°C | Same |
| Conjugate volume | 90 μL | Same |
| Development solution volume | 90 μL | Same |
| Stop solution volume | 200 μL | Same |
| Assay set-up | Random access | Same |
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| Similarities | | |
| --- | --- | --- |
| Item | Predicate Device
Phadia 250 | Test Device
Phadia 2500/5000 |
| Reagent packaging size | Various/Common | Various/Common
Introduction of new article number for EliA Sample Diluent (83-1071-01) is only due to larger filling volume. |
| Onboard storage of reagents | Yes | Same |
| Time to 1st result | ~2 hours | Same |
| Differences | | |
| --- | --- | --- |
| Item | Predicate Device
Phadia 250 | Test Device
Phadia 2500/5000 |
| Sample matrix | Human serum or plasma
(heparin, EDTA, citrate) | Human serum or plasma (Li-heparin, EDTA); i.e. citrate plasma is omitted for all tests |
| Instrument Platform | Phadia 250 (ImmunoCAP 250) | Phadia 2500 / 5000 |
| Measuring Range | EliA CENP: 0.4 – 240 U/mL
EliA U1RNP: 0.3 – 240 U/mL
EliA RNP70: 0.3 – 240 U/mL | EliA CENP: 0.7 – 240 U/mL
EliA U1RNP: 1.1 – 240 U/mL
EliA RNP70: 0.9 – 240 U/mL |
| Daily throughput | ~250 tests | Phadia 2500: ~2500 tests
Phadia 5000: ~5000 tests |
| Sample dilution | Phadia 250 uses a steel pipette to dilute the samples in Dilution Plates | Phadia 2500/5000 uses disposable Pipette Tips in Racks for pipetting samples in Dilution Well |
| Risk for carry-over | The warning “DO NOT REUSE” in the Phadia 250 DFU for EliA Conjugates is due to the fact that a low risk of conjugate contamination by carry-over from samples was identified. In order to reduce the risk, the single use statement for the conjugate was included in the Phadia 250 DFU. | When running EliA tests on the Phadia 2500/5000 instruments, there is no need for this warning statement because these instruments use disposable tips for pipetting samples and a separate pipette for the conjugate, and carry-over from samples to conjugate is impossible. |
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| Differences | | |
| --- | --- | --- |
| Item | Predicate Device
Phadia 250 | Test Device
Phadia 2500/5000 |
| Loading of
EliA carriers | The EliA carriers are loaded manually on the Loading Tray from where they can be processed directly or transferred to the cooled storage compartment. | The Phadia 2500/5000 instruments do not have such a Loading Tray. The EliA carriers are loaded into racks which are directly transferred to the cooled storage compartment |
| Barcode reader | The Phadia 250 instrument has a built-in barcode reader at the front of the instrument, but the operator needs to scan the barcodes manually by showing the reagents to the barcode reader. Alternatively, the operator can also enter the characters below the barcode manually. | The Phadia 2500/5000 instruments dispose of a built-in barcode reader, and the reagents are on a moving belt which conveys them past the barcode reader. The lot-specific information will be read automatically by the instrument during loading. |
| Process time /
Time to patient result | Phadia 250 needs one minute to process one well and provides the results at a 1-minute interval. | Phadia 2500/5000 processes two wells in parallel in 48 seconds and provides the results at a 24-second interval. |
K. Standard/Guidance Documents Referenced:
- CLSI EP05-A3: Evaluation of Precision Performance of Quantitative Measurement Methods; September 2014
- CLSI EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; April 2003
- CLSI EP17-A: Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline
L. Test Principle:
EliA tests are fluorescence 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 EliA wells are coated with human recombinant centromere protein B, human recombinant RNP (RNP70, A, C) or with human recombinant RNP (70 kDa) protein. If present in the patient's specimen, antibodies to these proteins bind to 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-
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bound conjugate is washed away, and the bound complex is incubated with a Development Solution. After stopping the reaction, the fluorescence in the reaction mixture is measured. The higher the value of fluorescent signal, 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:
### 1. Analytical performance:
All results presented below met the manufacturer’s pre-determined acceptance criteria for all analytical performance studies.
#### a. Precision/Reproducibility:
To determine the precision of the assay, the variability was assessed in a study with a total of 21 runs (three instruments total and seven runs per instrument). The study was performed with a single run/day over a period of seven days. Each sample was tested in four replicates/run giving in total 84 replicates per sample (3 instruments × 7 runs x 4 replicates = 84). The data were calculated against the calibration curve from Day 1. Five native patient serum samples were tested: one negative, one or two in the equivocal range, and two or three positive specimens. All samples were tested on all instruments. The results are summarized in the tables below:
EliA CENP on Phadia 2500/5000:
| Sample | Mean (EliA U/mL) | Within-Run | | Between-Run | | Between-instrument | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 1 | 1.1 | 0.1 | 9.9 | 0.1 | 6.1 | 0.2 | 15.8 | 0.2 | 19.6 |
| 2 | 7.1 | 0.4 | 5.8 | 0.3 | 4.9 | 0.3 | 3.9 | 0.6 | 8.6 |
| 3 | 11.4 | 0.4 | 3.5 | 0.4 | 3.9 | 0.5 | 4.2 | 0.8 | 6.7 |
| 4 | 129.8 | 4.3 | 3.3 | 5.1 | 3.9 | 3.9 | 3.0 | 7.7 | 6.0 |
| 5 | 229.7 | 9.8 | 4.3 | 15.7 | 6.8 | 10.1 | 4.4 | 21.1 | 9.2 |
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EliA U1RNP on Phadia 2500/5000:
| Sample | Mean (EliA U/mL) | Within-Run | | Between-Run | | Between-instrument | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 1 | 2.2 | 0.4 | 17.9 | 0.3 | 12.5 | 0.2 | 11.0 | 0.5 | 24.4 |
| 2 | 5.7 | 0.3 | 5.4 | 0.2 | 2.8 | 0.1 | 1.8 | 0.4 | 6.4 |
| 3 | 10.4 | 0.5 | 4.7 | 0.4 | 4.2 | 0.6 | 5.6 | 0.9 | 8.5 |
| 4 | 135.4 | 5.2 | 3.8 | 7.2 | 5.3 | 6.5 | 4.8 | 11.0 | 8.1 |
| 5 | 231.7 | 9.1 | 3.9 | 9.0 | 3.9 | 10.7 | 4.6 | 16.7 | 7.2 |
EliA RNP70 on Phadia 2500/5000:
| Sample | Mean (EliA U/mL) | Within-Run | | Between-Run | | Between-instrument | | Total Imprecision | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 1 | 1.4 | 0.1 | 5.4 | 0.1 | 4.4 | 0.2 | 17.0 | 0.3 | 18.4 |
| 2 | 7.9 | 0.2 | 2.9 | 0.3 | 4.4 | 0.5 | 6.2 | 0.6 | 8.1 |
| 3 | 8.9 | 0.5 | 5.2 | 0.5 | 5.5 | 0.5 | 5.2 | 0.8 | 9.2 |
| 4 | 93.1 | 5.1 | 5.5 | 5.5 | 5.9 | 2.9 | 3.1 | 8.0 | 8.6 |
| 5 | 190.2 | 9.4 | 5.0 | 10.3 | 5.4 | 10.6 | 5.6 | 17.5 | 9.2 |
b. Linearity/assay reportable range:
Four patient serum samples were diluted in EliA Sample Diluent and tested with one lot of EliA CENP, EliA U1RNP and EliA RNP70 and one set of system reagents on Phadia 2500/5000. The results of the dilutions were compared with their expected values. The ratio observed/expected (O/E) was calculated. Mean observed value was used in the calculation. The results of the regression analysis are summarized in the tables below:
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EliA CENP on Phadia 2500/5000:
| Sample | Dilution Range (EliA U/mL) | Slope (95% CI) | Intercept (95% CI) | R² | %CV Range |
| --- | --- | --- | --- | --- | --- |
| 1 | 0.5–24.3 | 1.00 (0.99–1.01) | 0.17 (0.02–0.31) | 1.00 | 0.3–3.9 |
| 2 | 2.1–180.3 | 1.01 (0.99–1.02) | 0.10 (-0.83–1.03) | 1.00 | 0.4–6.3 |
| 3 | 3.8–305.0 | 1.04 (0.99–1.10) | 2.70 (-4.21–9.62) | 0.99 | 0.7–8.0 |
| 4 | 0.4–18.8 | 0.99 (0.97–1.02) | -0.22 (-0.44–0.00) | 1.00 | 1.3–4.3 |
The reportable range (Limit of Detection to upper limit) for EliA CENP is from 0.4 to 240 EliA U/mL. The measuring range (Limit of Quantitation to upper limit) is from 0.7 to 240 EliA U/mL.
EliA U1RNP on Phadia 2500/5000:
| Sample | Dilution Range (EliA U/mL) | Slope (95% CI) | Intercept (95% CI) | R² | %CV Range |
| --- | --- | --- | --- | --- | --- |
| 1 | 0.5–18.0 | 1.01 (0.99–1.02) | 0.08 (-0.04–0.20) | 1.00 | 1.2–20.8 |
| 2 | 0.6–51.3 | 1.01 (0.99–1.02) | 0.04 (-0.20–0.27) | 1.00 | 1.9–6.4 |
| 3 | 0.6–15.8 | 1.00 (0.96–1.04) | 0.28 (0.01–0.56) | 1.00 | 1.2–19.1 |
| 4 | 4.7–284.7 | 1.04 (0.98–1.10) | 1.86 (-4.75–8.47) | 0.99 | 1.3–9.9 |
The reportable range (Limit of Detection to upper limit) for EliA U1RNP is from 0.5 to 240 EliA U/mL. The measuring range (Limit of Quantitation to upper limit) is from 1.1 to 240 EliA U/mL.
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EliA RNP70 on Phadia 2500/5000:
| Sample | Dilution Range (EliA U/mL) | Slope (95% CI) | Intercept (95% CI) | R² | %CV Range |
| --- | --- | --- | --- | --- | --- |
| 1 | 0.6–62.5 | 1.02 (1.00–1.04) | 0.07 (−0.47–0.49) | 1.00 | 0.8–6.0 |
| 2 | 0.4–38.7 | 1.01 (0.98–1.04) | −0.42 (−0.87–0.02) | 1.00 | 0.6–10.0 |
| 3 | 6.9–251.8 | 1.00 (0.97–1.03) | 3.07 (0.05–6.09) | 1.00 | 0.9–4.1 |
| 4 | 3.7–294.6 | 1.01 (0.98–1.03) | 3.38 (0.39–6.38) | 1.00 | 1.4–7.7 |
The reportable range (Limit of Detection to upper limit) for EliA RNP70 is from 0.3 to 240 EliA U/mL. The measuring range (Limit of Quantitation to upper limit) is from 0.9 to 240 EliA U/mL.
The following statements are included in the package inserts: "Please note that concentration values between LoD and LoQ may show a higher uncertainty." and "Please note that due to differing binding characteristics of the antibodies in patient samples, not all sera can be diluted linearly within the measuring range."
The hook effect was previously reviewed in K082759 (EliA CENP and EliA U1RNP) and K083117 (EliA RNP70).
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Same as in the predicate devices. Refer to K082759 (EliA CENP and EliA U1RNP) and K083117 (EliA RNP70).
d. Detection limit:
The limit of blank (LoB) and limit of detection (LoD) studies were performed on a Phadia 2500 instrument. LoB was measured in two assay runs with each run including a blank sample in 33 replicates, giving a total of 66 replicates. LoD was measured in two assay runs by testing three low level serum samples in 11 replicates per sample in a run, giving a total of 66 replicates. The limit of quantitation (LoQ) was based on 66 determinations and a precision target of 20%. The results are summarized in the table below:
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| Assay | LoB | LoD | LoQ |
| --- | --- | --- | --- |
| EliA CENP (EliA U/mL) | 0.2 | 0.4 | 0.7 |
| EliA U1RNP (EliA U/mL) | 0.2 | 0.5 | 1.1 |
| EliA RNP70 (EliA U/mL) | 0.2 | 0.3 | 0.9 |
e. Analytical specificity:
i) Interference:
Interference by endogenous substances was previously reviewed in K082759 (EliA CENP and EliA U1RNP) and K083117 (EliA RNP70).
ii) Cross-reactivity:
Cross-reactivity was previously reviewed in K082759 (EliA CENP and EliA U1RNP) and K083117 (EliA RNP70).
iii) Carry-over:
The Phadia 2500/5000 instruments use disposable tips for pipetting samples and a separate pipette for the conjugate, therefore carry-over from samples to conjugate was not evaluated.
f. Assay cut-off:
The assay cut-offs are the same as in the predicate devices (refer to K082759 and K083117) and are summarized in the tables below:
EliA CENP:
| < 7 EliA U/mL | Negative |
| --- | --- |
| 7–10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |
EliA U1RNP:
| < 5 EliA U/mL | Negative |
| --- | --- |
| 5–10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |
EliA RNP70:
| < 7 EliA U/mL | Negative |
| --- | --- |
| 7–10 EliA U/mL | Equivocal |
| > 10 EliA U/mL | Positive |
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2. Comparison studies:
a. Method comparison with predicate device:
See 2c. Instrument Comparison below.
b. Matrix comparison:
Matrix comparison between serum and plasma was reviewed in K141375K082759 (EliA CENP and EliA U1RNP) and K083117 (EliA RNP70).
c. Instrument comparison:
The purpose of this study was to evaluate conformance and show comparability of EliA CENP, EliA U1RNP, and EliA RNP70 on the Phadia 250 instrument versus the Phadia 2500/5000 instrument.
1) EliA CENP:
A total of 100 samples (75 positive, 17 negative and 8 equivocal) were run in singlicate on one Phadia 250 instrument and three Phadia 2500/5000 instruments for comparison. Only samples inside the measuring range were included in the calculations. Results were analyzed by Passing-Bablok regression and summarized below:
| Instrument | Range Tested (EliA U/mL) | Intercept (95% CI) | Slope (95% CI) | *R |
| --- | --- | --- | --- | --- |
| PH2500/5000 A | 1.7–232.9 | −0.30
(−0.55 to −0.07) | 1.02
(1.00–1.03) | 0.997 |
| PH2500/5000 B | 1.6–223.3 | 0.44
(0.07–0.78) | 0.98
(0.95–1.00) | 0.990 |
| PH2500/5000 C | 1.8–235.7 | −0.40
(−0.91 to −0.09) | 1.04
(1.03–1.07) | 0.995 |
| *Pearson correlation coefficient | | | | |
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Positive percent agreement (PPA), negative percent agreement (NPA), and total percent agreement (TPA) were evaluated for each instrument pair comparison with equivocal results considered positive in the table below:
| | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
| --- | --- | --- | --- |
| PPA
(95% CI) | 100.0%
(95.7%–100.0%) | 100.0%
(95.7%–100.0%) | 100.0%
(95.5%–100.0%) |
| NPA
(95% CI) | 94.1%
(71.3%–99.9%) | 88.2%
(63.6%–98.5%) | 88.2%
(63.6%–98.5%) |
| TPA
(95% CI) | 99.0%
(94.6%–100.0%) | 98.0%
(93.0%–99.8%) | 97.9%
(92.7%–99.7%) |
Results when equivocal results are considered negative are presented in the table below:
| | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
| --- | --- | --- | --- |
| PPA
(95% CI) | 98.7%
(92.8%–100.0%) | 98.7%
(92.8%–100.0%) | 98.6%
(92.5%–100.0%) |
| NPA
(95% CI) | 100.0%
(86.3%–100.0%) | 92.0%
(74.0%–99.0%) | 96.0%
(79.6%–99.9%) |
| TPA
(95% CI) | 99.0%
(94.6%–100.0%) | 97.0%
(91.5%–99.4%) | 97.9%
(92.7%–99.7%) |
No sample switched from negative to positive or vice versa. Three samples in the equivocal range were tested positive and three samples in the negative range were tested equivocal.
2) EliA U1RNP:
A total of 112 samples (77 positive, 17 negative and 18 equivocal) were run in singlicate on one Phadia 250 instrument and three Phadia 2500/5000 instruments for comparison. Only samples inside the measuring range were included in the calculations. Results were analyzed by Passing-Bablok regression and summarized below:
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| Instrument | Range Tested (EliA U/mL) | Intercept (95% CI) | Slope (95% CI) | *R |
| --- | --- | --- | --- | --- |
| PH2500/5000 A | 1.5–225.3 | 0.68
(0.34–1.02) | 0.93
(0.90–0.95) | 0.990 |
| PH2500/5000 B | 1.6–239.3 | 0.03
(–0.36–0.65) | 1.02
(0.98–1.05) | 0.995 |
| PH2500/5000 C | 1.6–198.2 | 0.98
(0.43–1.26) | 0.92
(0.90–0.94) | 0.998 |
| *Pearson correlation coefficient | | | | |
For each instrument pair comparison, PPA, NPA, and TPA were evaluated with equivocal results considered positive in the table below:
| | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
| --- | --- | --- | --- |
| PPA
(95% CI) | 100.0%
(96.2%–100.0%) | 98.9%
(94.3%–100.0%) | 96.8%
(91.0%–99.3%) |
| NPA
(95% CI) | 100.0%
(80.5%–100.0%) | 94.1%
(71.3%–99.5%) | 100.0%
(80.5%–100.0%) |
| TPA
(95% CI) | 100.0%
(96.8%–100.0%) | 98.2%
(93.7%–99.8%) | 97.3%
(92.4%–99.4%) |
Results when equivocal results are considered negative are presented in the table below:
| | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
| --- | --- | --- | --- |
| PPA
(95% CI) | 98.7%
(93.0%–100.0%) | 97.4%
(90.9%–99.7%) | 100.0%
(95.3%–100.0%) |
| NPA
(95% CI) | 97.1%
(85.1%–99.9%) | 97.1%
(85.1%–99.9%) | 97.1%
(85.1%–99.9%) |
| TPA
(95% CI) | 98.2%
(93.7%–100.0%) | 97.3%
(92.4%–99.4%) | 99.1%
(95.1%–100.0%) |
No sample switched from negative to positive or vice versa. One samples in the negative range was tested equivocal and four samples in the equivocal range were tested negative. Two samples in the positive range were tested equivocal and one sample in the equivocal range was tested positive.
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3) EliA RNP70:
A total of 113 samples (75 positive, 26 negative and 12 equivocal) were run in singlicate on one Phadia 250 instrument and three Phadia 2500/5000 instruments for comparison. Only samples inside the measuring range were included in the calculations. Results were analyzed by Passing-Bablok regression and summarized below:
| Instrument | Range Tested (EliA U/mL) | Intercept (95% CI) | Slope (95% CI) | *R |
| --- | --- | --- | --- | --- |
| PH2500/5000 A | 0.5–238.8 | 0.35
(0.17–0.52) | 1.00
(0.98–1.02) | 0.995 |
| PH2500/5000 B | 0.4–236.4 | 0.29
(0.17–0.48) | 1.04
(1.01–1.06) | 0.992 |
| PH2500/5000 C | 0.5–234.6 | 0.33
(0.16–0.69) | 1.03
(1.01–1.06) | 0.993 |
| *Pearson correlation coefficient | | | | |
For each instrument pair comparison, PPA, NPA, and TPA were evaluated with equivocal results considered positive in the table below:
| | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
| --- | --- | --- | --- |
| PPA
(95% CI) | 100.0%
(95.8%–100.0%) | 100.0%
(95.8%–100.0%) | 100.0%
(95.8%–100.0%) |
| NPA
(95% CI) | 88.5%
(69.8%–97.6%) | 80.8%
(60.6%–93.4%) | 84.6%
(65.1%–95.6%) |
| TPA
(95% CI) | 97.3%
(92.3%–99.4%) | 95.5%
(89.8%–98.5%) | 96.4%
(91.1%–99.0%) |
Results when equivocal results are considered negative are presented in the table below:
| | PH2500/5000 A | PH2500/5000 B | PH2500/5000 C |
| --- | --- | --- | --- |
| PPA
(95% CI) | 98.6%
(92.6%–100.0%) | 100.0%
(95.1%–100.0%) | 98.6%
(92.7%–100.0%) |
| NPA
(95% CI) | 97.4%
(86.2%–99.9%) | 92.1%
(78.6%–98.3%) | 92.1%
(78.6%–98.3%) |
| TPA
(95% CI) | 98.2%
(93.6%–99.8%) | 97.3%
(92.3%–99.4%) | 96.4%
(91.1%–99.0%) |
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No sample switched from negative to positive or vice versa. Five samples in the negative range were tested equivocal. One sample in the positive range was tested equivocal and three samples in the equivocal range were tested positive.
3. Clinical studies:
a. Clinical Sensitivity and Specificity:
Clinical performance values were reviewed in K082759 and K083117.
b. Other clinical supportive data (when a. is not applicable):
Not Applicable
4. Clinical cut-off:
Same as assay cut-off.
5. Expected values/Reference range:
The frequency distribution for CENP, U1RNP and RNP70 antibodies was investigated in a group of apparently healthy subjects (N=400) equally distributed by age and gender, using sera from a Caucasian population obtained from a blood bank. None of test samples was within the equivocal or the positive range in the three EliA assays. The test results are given in the table below and all measurements are shown in EliA U/mL:
| Test | N | Mean | Median | Range | 95^{th} Percentile | 99^{th} Percentile |
| --- | --- | --- | --- | --- | --- | --- |
| EliA CENP on Phadia 2500/5000 | 400 | 0.7 | 0.7 | 0.1–5.7 | 1.0 | 1.3 |
| EliA U1RNP on Phadia 2500/5000 | 400 | 1.5 | 1.5 | 0.4–3.9 | 2.2 | 3.0 |
| EliA RNP70 on Phadia 2500/5000 | 400 | 1.4 | 1.4 | 0.1–3.7 | 2.2 | 2.5 |
N. Proposed Labelling:
The labelling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
