(423 days)
No
The summary mentions image processing and automated microscopy but does not explicitly mention or describe the use of AI or ML algorithms for image analysis or interpretation. The interpretation of digital images is still stated to be reviewed and interpreted from the computer monitor, and results generated with the NOVA View device must be confirmed by a trained operator.
No
This device is an in vitro diagnostic (IVD) tool used for the qualitative and/or semi-quantitative determination of anti-double stranded DNA (dsDNA) IgG antibodies to aid in the diagnosis of systemic lupus erythematosus (SLE). It does not directly treat or prevent a disease, which are characteristics of a therapeutic device.
Yes
The intended use explicitly states that the device aids in the diagnosis of systemic lupus erythematosus (SLE) by determining anti-double stranded DNA (dsDNA) IgG antibodies in human serum.
No
The device is a kit that includes reagents and slides for an indirect immunofluorescence assay, which are physical components. While it utilizes software for image interpretation, it is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it's an "indirect immunofluorescent assay for the qualitative and/or semi-quantitative determination of anti-double stranded DNA (dsDNA) IgG antibodies in human serum". This is a test performed on a sample taken from the human body (serum) to provide information about a medical condition (presence of anti-dsDNA antibodies to aid in the diagnosis of SLE).
- Device Description: The description details a laboratory procedure involving human serum samples, reagents, and a method for detecting antibodies. This aligns with the definition of an in vitro diagnostic test.
- Performance Studies: The document includes extensive performance studies (Precision, Reproducibility, Linearity, Interference, Sample Stability, Reagent Stability, Clinical performance characteristics, Comparison with predicate device, SWT validation) which are standard requirements for demonstrating the analytical and clinical validity of an IVD.
- Predicate Device: The mention of a predicate device (K880742; NOVA Lite ™ DSDNA) further indicates that this device is being compared to an already cleared IVD.
All these elements strongly support the classification of this device as an In Vitro Diagnostic.
N/A
Intended Use / Indications for Use
NOVA Lite® DAPI dsDNA Crithidia luciliae is an indirect immunofluorescent assay for the qualitative and/or semi-quantitative determination of anti-double stranded DNA (dsDNA) IgG antibodies in human serum by NOVA View Automated Fluorescence Microscope or manual fluorescence microscopy. The presence of anti-dsDNA can be used in conjunction with other serological and clinical findings to aid in the diagnosis of systemic lupus erythematosus (SLE). All results generated with NOVA View device must be confirmed by a trained operator.
Product codes
KTL, PIV
Device Description
The NOVA Lite DAPI dsDNA Crithidia luciliae Kit is an indirect immunofluorescence assay for the qualitative detection and semi-quantitative determination of Anti-dsDNA Antibodies (IgG) in human serum.
Samples are diluted 1:10 in PBS and incubated with the antigen substrate (dsDNA on glass microscope slides). After incubation, unbound antibodies are washed off. The substrate is then incubated with antihuman IgG-FITC conjugate. The conjugate contains a DNA-binding blue fluorescent dye, 4',6-diamidino-2phenylindole (DAPI) that is required for NOVA View use. The blue dye is not visible by traditional fluorescence microscope at the wavelength where FITC fluorescence is viewed. Unbound reagent is washed off. Stained slides are read by manual fluorescence microscope or scanned with the NOVA View Automated Fluorescence Microscope. The resulting digital images are reviewed and interpreted from the computer monitor. dsDNA positive samples exhibit an apple green fluorescence corresponding to areas of the substrate where autoantibody has bound.
Manual interpretation:
A sample is considered positive if specific kinetoplast plus nuclear staining is observed to be greater than the negative control.
NOVA View interpretation:
When slides are analyzed by NOVA View, digital images of representative fields of view of the well are captured. These digital images must be reviewed and interpreted from the computer monitor by a trained operator. At the same time when digital images are taken, NOVA View measures the FITC light intensity of the cells that are included in the region. NOVA View reports the measured fluorescence intensity in units of Light Intensity Units (LIU).
NOVA View provides the trained operator with the acquired digital images and the following supportive information:
- LIU value
- Negative/positive/indeterminate classification
NOVA View Single Well Titer (SWT):
The Single Well Titer (SWT) is a software application that estimates the endpoint titer (i.e. the highest dilution that produces positive result) for wells with a positive reaction, based on the obtained LIU.
The NOVA Lite® DAPI dsDNA Crithidia luciliae Kit contains the following:
- dsDNA Crithidia luciliae Slides; 12 wells/slide, with desiccant
- FITC IgG Conjugate with DAPI, containing 0.09% sodium azide; ready to use.
- Positive Control: dsDNA; human serum with antibodies to dsDNA antigen, containing 0.09% sodium azide; pre-diluted, ready to use.
- Negative Control: IFA System Negative Control, diluted human serum with no dsDNA antibodies present, containing 0.09% sodium azide; pre-diluted, ready to use.
- PBS II (40x) Concentrate, sufficient for making 2000 mL of 1x PBS II.
- Mounting Medium, containing 0.09% sodium azide
- Coverslips
Mentions image processing
Yes
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Fluorescence microscopy
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Trained operator / Not Found (Clinical Laboratories implied)
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Precision Study:
- Sample Size: 2 negative and 2 borderline samples, tested in triplicate, in 14 runs (2 runs per day) for 7 days, resulting in 42 data points for each sample.
- Data Source: Not specified, internal samples.
- Annotation Protocol: Slides were read with NOVA View, and digital images were interpreted by the operator. Manual reading was also performed. Results were categorized as NOVA View software interpretation, digital image reading results, and manual reading results.
Reproducibility Studies (Between sites/instruments reproducibility):
- Sample Size: Ten samples (3 negative, 7 positive), tested in three replicates, twice a day for 5 days at each site (30 data points per sample).
- Data Source: Not specified, likely internal or characterized samples.
- Annotation Protocol: Manual and digital reading was performed by two operators at each site to assess between operator reproducibility.
Reproducibility between lots:
- Sample Size: Twenty clinically and/or analytically characterized samples, tested in duplicate.
- Data Source: Not specified, likely characterized samples.
- Annotation Protocol: NOVA view output, digital image reading, and manual image reading were compared for qualitative agreement and grade agreement.
Linearity Study:
- Sample Size: 3 positive samples (one high positive, one medium positive and one low positive).
- Data Source: Not specified, likely characterized samples.
- Annotation Protocol: Samples were serially diluted from 1:10 to 1:5120. Assessed with the NOVA Lite DAPI dsDNA Crithidia luciliae Kit and read with the NOVA View, digital images were interpreted and confirmed. All slides were read with manual microscopy. Qualitative and semi-quantitative results (using a scale of 0 (negative) to 4 (strong positive)) were captured for the manual and digital reading.
Interference Study:
- Sample Size: Three specimens (one negative, one positive, one strong positive) and various interfering substances.
- Data Source: Not specified, likely characterized samples.
- Annotation Protocol: Interfering substances (hemoglobin, bilirubin, triglycerides, cholesterol, Rituximab, Methylprednisolone, Cyclophosphamide, Methotrexate, Azathioprine, Ibuprofen, Naproxen, Hydroxychloroquine, Mycophenolate) were spiked into every specimen at three different concentrations in 10% of total specimen volume. Rheumatoid factor (RF) positive samples were added at 10%, 30%, and 50% (volume). Resulting samples were assessed in triplicates with the NOVA Lite DAPI dsDNA Crithidia luciliae Kit and read with NOVA View. Digital images were interpreted and confirmed. All slides were read by the same operator with manual microscopy.
Sample Stability and Handling:
- Sample Size: Three samples (negative, around the cut-off, and positive).
- Data Source: Not specified, likely characterized samples.
- Annotation Protocol: Samples were tested in duplicates for up to 21 days while stored at 2-8°C, up to 48 hours while stored at room temperature, and after repeated freeze/thaw cycles (up to 3 cycles). Results compared to control samples (day zero, stored at 2-8°C). NOVA View, manual reading, and digital image interpretation were performed.
Reagent Stability (Accelerated Stability Studies):
- Sample Size: Components of the NOVA Lite DAPI dsDNA Crithidia luciliae Kit (Slides, conjugate, Negative control, positive control, PBSII and Mounting Medium) from multiple lots.
- Data Source: Not specified, likely manufactured lots.
- Annotation Protocol: New sealed kits were placed in an incubator at 37°C ± 3°C for up to 4 weeks. All components were tested at the end of the experiment together with a control stored at 5 ± 3°C. Reactivity of the kits was calculated by comparing results to the control. Manual and digital methods were used.
Reagent Stability (In-use stability - Conjugate and Controls):
- Sample Size: Conjugate and control bottles.
- Data Source: Not specified, likely manufactured lots.
- Annotation Protocol: Opened conjugate and control bottles were stored in the refrigerator and tested each week against unopened bottles using positive and negative controls on dsDNA Crithidia luciliae kits.
Real time stability:
- Sample Size: Lot 1, Lot 2, and Lot 3 of the NOVA Lite DAPI dsDNA Crithidia luciliae Kit.
- Data Source: Not specified, likely manufactured lots.
- Annotation Protocol: Results were available up to 24 months for lot 1, 15 months for lot 2, and 19 months for lot 3. Ongoing study to verify 2-year expiration where results are expected to fall within their respective ranges.
Clinical Performance Characteristics (Clinical sensitivity, specificity):
- Sample Size: 766 clinically characterized serum samples.
- Data Source: Inova Diagnostics. Distribution of cohorts by diagnosis is provided in a table.
- Annotation Protocol: NOVA View, manual reading, and digital reading were performed. Sensitivity (on SLE) and specificity were calculated on the combined population.
Clinical studies 3 sites:
- Sample Size: 269 clinically characterized samples tested at three sites (total samples: 807).
- Data Source: Clinical sites.
- Annotation Protocol: Samples were tested using the NOVA Lite DAPI dsDNA (Crithidia luciliae) Kit according to its direction insert. Clinical Sensitivity/Specificity for NOVA View, Manual Reading, and Digital Reading were determined. Correlation to clinical diagnosis for each mode was assessed at each site.
Comparison with predicate device:
- Sample Size: 744 serum samples (comprising 391 serum samples from patients with SLE and 353 samples from patients with other diseases). This is a subset of the samples used in the clinical study.
- Data Source: Not specified, previously characterized samples from the clinical study.
- Annotation Protocol: Method comparison was performed with results obtained with the NOVA Lite DAPI dsDNA Crithidia luciliae Kit by NOVA View software interpretation and with the predicate device.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision Study:
- Study Type: Analytical precision.
- Sample Size: 6 samples (2 negative, 2 borderline, 2 positive), 42 data points per sample.
- Key Results: For both digital images reading, grades were within ± one reactivity grade within one run (within triplicates), and the average grade was no more than one reactivity grade different between runs.
Reproducibility Studies (Between sites/instruments reproducibility):
- Study Type: Analytical reproducibility between sites/instruments and operators.
- Sample Size: 10 samples (3 negative, 7 positive), 30 data points per sample per site.
- Key Results: Manual Reading: Overall agreement between operators (per site) was 99.7% for Site 1, 100.0% for Site 2, and 100.0% for Site 3. Digital Reading: Overall agreement between operators (per site) was 100.0% for Site 1, 100.0% for Site 2, and 100.0% for Site 3.
Reproducibility between lots:
- Study Type: Analytical Lot-to-lot reproducibility.
- Sample Size: 20 clinically and/or analytically characterized samples.
- Key Results:
- Qualitative Agreement:
- NOVA View: Negative agreement (96.4%), Positive agreement (91.7%), Total agreement (95.0%).
- Manual: Negative agreement (100.0%), Positive agreement (100.0%), Total agreement (100.0%).
- Digital: Negative agreement (100.0%), Positive agreement (92.9%), Total agreement (97.5%).
- Grade Agreement:
- Manual: 100% were within ±1 grade.
- Digital: Most grades were within ±1 grade (two comparisons 100%, one comparison 98%).
- Qualitative Agreement:
Linearity:
- Study Type: Analytical linearity.
- Sample Size: 3 positive samples (serially diluted).
- Key Results:
- High Positive sample: Manual titer 640, Digital titer 640.
- Medium Positive sample: Manual titer 20, Digital titer 40.
- Low Positive sample: Manual titer 20, Digital titer 20.
Interference:
- Study Type: Analytical interference study.
- Sample Size: 3 specimens (negative, positive, strong positive) with various interferents.
- Key Results: No interference was detected with specified concentrations of hemoglobin, bilirubin, triglycerides, cholesterol, rheumatoid factor, azathioprine, cyclophosphamide, hydroxychloroquine, ibuprofen, methotrexate, methylprednisolone, mycophenolate, naproxen, rituximab, and belimumab.
Sample Stability and Handling:
- Study Type: Analytical sample stability.
- Sample Size: 3 samples (negative, cut-off, positive).
- Key Results: All samples fulfilled acceptance criteria (results within ±1 grade, no category change) for storage up to 48 hours at room temperature, up to 7 days at 2-8°C, and up to 3 freeze/thaw cycles.
Reagent Stability:
- Study Type: Analytical accelerated and in-use reagent stability.
- Sample Size: Multiple lots of the kit components; conjugate and control bottles.
- Key Results: Acceptance criteria for two-year preliminary expiration dating were met for accelerated stability. All samples tested against the control kit (week 0) were within ±1 reactivity grade. Conjugate and controls were stable for 8 weeks after opening when stored at 2-8℃. Real-time stability results were within acceptance limits up to 24 months for Lot 1, 15 months for Lot 2, and 19 months for Lot 3.
Clinical sensitivity, specificity (initial clinical study):
- Study Type: Clinical Sensitivity and Specificity.
- Sample Size: 766 clinically characterized serum samples.
- Key Results:
- Manual: Sensitivity 48.1% (95% CI: 43.2-53.0), Specificity 91.2% (95% CI: 87.9-93.7).
- Digital: Sensitivity 48.1% (95% CI: 43.2-53.0), Specificity 92.3% (95% CI: 89.1-94.6).
- NOVA View: Sensitivity 57.0% (95% CI: 52.1-61.8), Specificity 88.8% (95% CI: 85.2-91.6).
Clinical studies 3 sites:
- Study Type: Multi-site Clinical Sensitivity and Specificity.
- Sample Size: 807 total samples (269 samples tested at 3 sites).
- Key Results:
- All sites combined (N=807):
- NOVA View: Sensitivity 40.0% (95% CI: 34.6-45.6), Specificity 85.4% (95% CI: 82.1-88.2).
- Manual Reading: Sensitivity 32.7% (95% CI: 27.6-38.2), Specificity 95.5% (95% CI: 93.3-97.0).
- Digital Reading: Sensitivity 34.0% (95% CI: 28.9-39.5), Specificity 95.5% (95% CI: 93.3-97.0).
- All sites combined (N=807):
Comparison with predicate device:
- Study Type: Method Comparison.
- Sample Size: 744 serum samples.
- Key Results:
- NOVA Lite DAPI dsDNA manual vs. Predicate device manual: Positive agreement 87.8%, Negative agreement 96.0%, Total agreement 93.7%.
- NOVA Lite DAPI dsDNA manual vs. Predicate device digital: Positive agreement 88.0%, Negative agreement 94.6%, Total agreement 92.7%.
- NOVA Lite DAPI dsDNA manual vs. Predicate device NOVA View: Positive agreement 87.0%, Negative agreement 85.3%, Total agreement 85.7%.
- Grade agreement (NOVA Lite DAPI dsDNA manual vs. Predicate device manual): +/- 2 reactivity grade = 99.6%.
- Grade agreement (NOVA Lite DAPI dsDNA manual vs. Predicate device digital): +/- 2 reactivity grade = 98.4%.
SWT Validation:
- Study Type: Software application validation.
- Sample Size: 31 positive samples.
- Key Results:
- SWT within ± 1 dilution step of Manual end-point: 80.6%.
- SWT within ± 1 dilution step of Digital end-point: 83.9%.
- SWT within ± 2 dilution steps of Manual end-point: 93.5%.
- SWT within ± 2 dilution steps of Digital end-point: 93.5%.
- In between sites reproducibility study: 100% (14 out of 14) of SWT results at two external sites were within ± 1 dilution step of manual titer, and 92.9% (13 out of 14) were within ± 1 dilution step of digital titer; 100% of SWT results were within ± 2 dilution steps of both manual and digital titer.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Clinical sensitivity, specificity (initial clinical study):
- Manual: Sensitivity 48.1% (95% CI: 43.2-53.0), Specificity 91.2% (95% CI: 87.9-93.7)
- Digital: Sensitivity 48.1% (95% CI: 43.2-53.0), Specificity 92.3% (95% CI: 89.1-94.6)
- NOVA View: Sensitivity 57.0% (95% CI: 52.1-61.8), Specificity 88.8% (95% CI: 85.2-91.6)
Clinical studies 3 sites (All sites combined, N=807):
- NOVA View: Sensitivity 40.0% (95% CI: 34.6-45.6), Specificity 85.4% (95% CI: 82.1-88.2)
- Manual Reading: Sensitivity 32.7% (95% CI: 27.6-38.2), Specificity 95.5% (95% CI: 93.3-97.0)
- Digital Reading: Sensitivity 34.0% (95% CI: 28.9-39.5), Specificity 95.5% (95% CI: 93.3-97.0)
Comparison with predicate device (N=744):
- 708205 manual vs 708215 manual: Positive agreement 87.8% (95% CI: 82.7-91.5), Negative agreement 96.0% (95% CI: 94.0-97.4), Total agreement 93.7%
- 708205 manual vs 708215 digital: Positive agreement 88.0% (95% CI: 82.9-91.7), Negative agreement 94.6% (95% CI: 92.3-96.2), Total agreement 92.7%
- 708205 manual vs 708215 NOVA View: Positive agreement 87.0% (95% CI: 81.8-90.9), Negative agreement 85.3% (95% CI: 82.0-88.0), Total agreement 85.7%
Expected values in healthy subjects (N=120):
- Manual interpretation: 3.3% positive results
- NOVA View software interpretation: 9.2% positive results
- Digital interpretation: 0.8% positive results
Predicate Device(s)
NOVA Lite ™ DSDNA, 510(k) number: K880742
Reference Device(s)
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 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).
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December 11, 2020
Inova Diagnostics, Inc. Carolina Auza Supervisor, Research & Development 9900 Old Grove Rd San Diego, California 92131
Re: K192916
Trade/Device Name: NOVA Lite DAPI dsDNA Crithidia luciliae Kit Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear Antibody Immunological Test System Regulatory Class: Class II Product Code: KTL, PIV Dated: November 11, 2020 Received: November 12, 2020
Dear Carolina Auza:
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 statutes and regulations administered by other Federal agencies. You must comply with all the Act's
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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 (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-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 (Katelin) Mao, Ph.D. Acting Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health 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) K192916
Device Name NOVA Lite DAPI dsDNA Crithidia luciliae Kit
Indications for Use (Describe)
NOVA Lite® DAPI dsDNA Crithidia luciliae is an indirect immunofluorescent assay for the qualitative and/or semiquantitative determination of anti-double stranded DNA (dsDNA) IgG antibodies in human serum by NOVA View Automated Fluorescence Microscope or manual fluorescence microscopy. The presence of anti-dsDNA can be used in conjunction with other serological and clinical findings to aid in the diagnosis of systemic lupus erythematosus (SLE). All results generated with NOVA View device must be confirmed by a trained operator.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| X Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary
NOVA Lite® DAPI dsDNA Crithidia luciliae Kit
Page 1 of 19
Table of Contents
| Administrative data |
|---|
| Predicate device |
| Device description |
| Intended use(s) |
| Indications for use…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… |
| Substantial equivalence |
| Comparison to predicate device |
| Analytical performance characteristics |
| Nomenclature used in the studies |
| Precision |
| Reproducibility Studies |
| Linearity |
| Interference |
| Sample Stability and Handling |
| Reagent Stability |
| Cut-off, reference range |
| Clinical performance characteristics |
| Clinical sensitivity, specificity |
| Expected values |
| Comparison with predicate device |
| System description |
This summary of the 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
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Administrative data
| Submitter: | Inova Diagnostics, Inc.
9900 Old Grove Road,
San Diego, CA, 92131 |
|-------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Purpose of submission: | New device(s) |
| Devices in the submission: | NOVA Lite® DAPI dsDNA Crithidia luciliae Kit |
| Scientific contact: | Carolina Auza, Supervisor, Research and Development
Inova Diagnostics, Inc.
9900 Old Grove Road, San Diego, CA, 92131
Phone: 858-586-9900 x 1345
Fax: 858-863-0025
email: cauza@inovadx.com |
| Quality Systems contact: | Ronda Elliott, VP, Quality Systems and RA
Inova Diagnostics, Inc.
9900 Old Grove Road, San Diego, CA, 92131
Phone: 858-586-9900 x 1381
Fax: 858-863-0025
email: relliott@inovadx.com |
| Device name (assay kit): | Proprietary name: NOVA Lite® DAPI dsDNA Crithidia luciliae Kit
Common name: anti-double stranded DNA (dsDNA) kit
Classification name: test system, anti-double stranded DNA (dsDNA) antibodies |
| Regulation Description: | Antinuclear antibody immunological test system |
| Regulation Medical Specialty: | Immunology |
| Review Panel: | Immunology |
| Product Code: | KTL
PIV |
| Classification Panel: | 82- Immunology |
| Regulation Number: | 866.5100- Antinuclear Antibody Immunological Test System |
5
Predicate device
NOVA Lite ™ DSDNA, 510(k) number: K880742
Device description
The NOVA Lite DAPI dsDNA Crithidia luciliae Kit is an indirect immunofluorescence assay for the qualitative detection and semi-quantitative determination of Anti-dsDNA Antibodies (IgG) in human serum.
Samples are diluted 1:10 in PBS and incubated with the antigen substrate (dsDNA on glass microscope slides). After incubation, unbound antibodies are washed off. The substrate is then incubated with antihuman IgG-FITC conjugate. The conjugate contains a DNA-binding blue fluorescent dye, 4',6-diamidino-2phenylindole (DAPI) that is required for NOVA View use. The blue dye is not visible by traditional fluorescence microscope at the wavelength where FITC fluorescence is viewed. Unbound reagent is washed off. Stained slides are read by manual fluorescence microscope or scanned with the NOVA View Automated Fluorescence Microscope. The resulting digital images are reviewed and interpreted from the computer monitor. dsDNA positive samples exhibit an apple green fluorescence corresponding to areas of the substrate where autoantibody has bound.
Manual interpretation
A sample is considered positive if specific kinetoplast plus nuclear staining is observed to be greater than the negative control.
NOVA View interpretation
When slides are analyzed by NOVA View, digital images of representative fields of view of the well are captured. These digital images must be reviewed and interpreted from the computer monitor by a trained operator. At the same time when digital images are taken, NOVA View measures the FITC light intensity of the cells that are included in the region. NOVA View reports the measured fluorescence intensity in units of Light Intensity Units (LIU).
NOVA View provides the trained operator with the acquired digital images and the following supportive information:
- LIU value
- Negative/positive/indeterminate classification
NOVA View Single Well Titer (SWT)
The Single Well Titer (SWT) is a software application that estimates the endpoint titer (i.e. the highest dilution that produces positive result) for wells with a positive reaction, based on the obtained LIU.
The NOVA Lite® DAPI dsDNA Crithidia luciliae Kit contains the following:
- dsDNA Crithidia luciliae Slides; 12 wells/slide, with desiccant
- FITC IgG Conjugate with DAPI, containing 0.09% sodium azide; ready to use.
- Positive Control: dsDNA; human serum with antibodies to dsDNA antigen, containing 0.09% sodium azide; pre-diluted, ready to use.
6
- Negative Control: IFA System Negative Control, diluted human serum with no dsDNA antibodies present, containing 0.09% sodium azide; pre-diluted, ready to use.
- PBS II (40x) Concentrate, sufficient for making 2000 mL of 1x PBS II.
- Mounting Medium, containing 0.09% sodium azide
- Coverslips
Intended use(s)
NOVA Lite® DAPI dsDNA Crithidia luciliae is an indirect immunofluorescent assay for the qualitative and/or semi-quantitative determination of anti-double stranded DNA (dsDNA) IgG antibodies in human serum by NOVA View Automated Fluorescence Microscope or manual fluorescence microscopy. The presence of anti-dsDNA can be used in conjunction with other serological and clinical findings to aid in the diagnosis of systemic lupus erythematosus (SLE). All results generated with NOVA View device must be confirmed by a trained operator.
Indications for use
Same as Intended use.
Substantial equivalence
The NOVA Lite DAPI dsDNA Crithidia luciliae reagents have the same intended use and assay principle as the predicate device NOVA Lite ™ DSDNA [510(k) K880742]
| Similarities | ||
|---|---|---|
| Item | NOVA Lite DAPI Crithidia luciliae Kit | Predicate Device |
| Intended use | NOVA Lite® DAPI dsDNA Crithidia luciliae is an indirect immunofluorescent assay for the qualitative and/or semi-quantitative determination of anti-double stranded DNA (dsDNA) IgG antibodies in human serum by NOVA View Automated Fluorescence Microscope or manual fluorescence microscopy. The presence of anti-dsDNA can be used in conjunction with other serological and clinical findings to aid in the diagnosis of systemic lupus erythematosus (SLE). All results generated with NOVA View device must be confirmed by a trained operator. | NOVA Lite dsDNA Crithidia luciliae is an indirect immunofluorescent assay for the screening and semi-quantitative determination of anti-double stranded DNA (dsDNA) in human serum. The presence of anti-double stranded DNA can be used in conjunction with other serological tests and clinical findings to aid in the diagnosis of systemic lupus erythematosus (SLE). |
Comparison to predicate device
7
| Similarities | ||
|---|---|---|
| Item | NOVA Lite DAPI Crithidia luciliae Kit | Predicate Device |
| Analyte | Anti-dsDNA Antibodies (IgG) in human serum | Anti-dsDNA Antibodies (IgG) in human serum |
| Assay methodology | indirect immunofluorescence assay | indirect immunofluorescence assay |
| Antigen | Crithidia luciliae cells | Crithidia luciliae cells |
| Sample type | Serum | Serum |
| Sample dilution | 1:10 | 1:10 |
| Controls | Two levels of controls: one negative, one positive (dsDNA Positive) | Two levels of controls: one negative, one positive (dsDNA Positive) |
| Storage | 2-8 °C | 2-8 °C |
| Shelf life | 24 months | 24 months |
| Differences | |||
|---|---|---|---|
| ltem | NOVA Lite DAPI Crithidia luciliae Kit | Predicate Device | |
| Interpretation | by manual fluorescence microscopy | ||
| or with the NOVA View device | by manual fluorescence microscopy | ||
| Conjugate | FITC conjugated anti-human IgG (Fc | ||
| specific) with added 4',6-diamidino- | |||
| 2-phenylindole (DAPI) | FITC conjugated anti-human IgG (Fc | ||
| specific) | |||
| Additional dye in | |||
| Conjugate | 4',6-diamidino-2-phenylindole | ||
| (DAPI) | None |
Analytical performance characteristics
Nomenclature used in the studies
All studies have been performed by interpreting the results with both manual microscopy and with the NOVA View system.
- . "Manual" and "Manual reading" refers to results obtained by reading the slides with traditional fluorescence microscope.
- . "NOVA View" refers to software reported results obtained with the NOVA View Automated Fluorescence Microscope, such as Light Intensity Units (LIU) and positive/indeterminate classification information.
- . "Digital", "Digital reading" and "Digital image" refers to results obtained by reading NOVA View generated images on the computer monitor.
For statistical calculations, a positive result is presented as "1", and a negative result is presented as "0″. Intensity of the staining is expressed in reactivity grades. Grade 0 is negative; grades 1-4 are weak to strong positive.
- 4+ Brilliant apple green fluorescence
- 3+ Bright apple green fluorescence
- 2+ Clearly distinguishable positive fluorescence
- 1+ Lowest specific fluorescence that enables the nuclear and/or cytoplasmic staining to be clearly differentiated from the background fluorescence
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Precision
To assess the precision performance of the NOVA Lite dsDNA Crithidia luciliae Kit results, a study was performed by processing 2 negative and 2 borderline samples with various intensities, in three replicates, in 14 runs (2 runs per day) for 7 days in triplicate resulting in 42 data points for each sample. The slides were read with NOVA view and digital images were interpreted by the operator as well as being read with manual microscope: i.e. three set of results were generated: NOVA view software interpretation, digital image reading results and manual reading results.
Acceptance criteria: Difference between reactivity grades within one run (between replicates) are within ± one reactivity grade. Average reactivity grade difference between any runs is within ± one reactivity grade.
Results: For both digital images reading, grades were within ± one reactivity grade within one run (within triplicates), and the average grade was no more than one reactivity grade different between runs.
| Expected Result | Obtained Result | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Sample | n | Negative/ | ||||||||
| Positive (grade) | NOVA View results | Manual results | Digital results | |||||||
| % Negative | % Positive | Grade range (0-4+) | % Negative | % Positive | Grade range (0-4+) | % Negative | % Positive | |||
| Sample 1 | 42 | Positive (3-4) | 7% | 93% | 3-4 | 0% | 100% | 4 | 0% | 100% |
| Sample 2 | 42 | Negative (0-1) | 100% | 0% | 0-1 | 98% | 2% | 0 | 100% | 0% |
| Sample 3 | 42 | Borderline (0-2) | 45% | 55% | 0-2 | 14% | 86% | 1-2 | 10% | 90% |
| Sample 4 | 42 | Borderline (0-2) | 100% | 0% | 1-2 | 0% | 100% | 0-2 | 69% | 31% |
| Sample 5 | 42 | Positive (1-3) | 64% | 36% | 1-3 | 0% | 100% | 1-2 | 14% | 86% |
| Sample 6 | 42 | Negative (0) | 90% | 10% | 0 | 100% | 0% | 0 | 100% | 0% |
The results are summarized in the table below.
Reproducibility Studies
Between sites/instruments reproducibility
Ten samples (3 negative, 7 positive) were tested in three replicates, twice a day for 5 days at each site (30 data points per sample). Manual and digital reading was performed by two operators at each site, to assess between operator reproducibility.
Acceptance criteria: 90% agreement between operators and between sites
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NOVA Lite® DAPI dsDNA Crithidia luciliae Kit
Grades for site 1:
| Digital Reactivity Grade | Manual Reactivity Grade | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample 1 | Sample 1 | |||||||||||
| Day | Run 1 | Run 2 | Run 1 | Run 2 | ||||||||
| 1 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 4 | 4 | |||
| 2 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 4 | 4 | |||
| 3 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 4 | 4 | |||
| 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 3 | 3 | |||
| 5 | 4 | 4 | 4 | 4 | 4 | 3 | 4 | 4 | 4 | |||
| Sample 2 | Sample 2 | |||||||||||
| 1 | 4 | 4 | 4 | 4 | 4 | 3 | 3 | 4 | 4 | |||
| 2 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 3 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 5 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| Sample 3 | Sample 3 | |||||||||||
| 1 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 2 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 3 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| 5 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | |||
| Sample 4 | Sample 4 | |||||||||||
| 1 | 3 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 2 | 2 | 3 | 3 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 3 | 2 | 3 | 3 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 4 | 2 | 3 | 3 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 5 | 2 | 3 | 3 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| Sample 5 | Sample 5 | |||||||||||
| 1 | 1 | 1 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 2 | 2 | 1 | 1 | 1 | 2 | 2 | 2 | 2 | 2 | |||
| 3 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 4 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | |||
| 5 | 2 | 2 | 1 | 1 | 2 | 2 | 1 | 2 | 2 | |||
| Sample 6 | Sample 6 | |||||||||||
| 1 | 1 | 1 | 1 | 1 | 2 | 2 | 2 | 2 | 2 | |||
| 2 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 2 | 1 | |||
| 3 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |||
| 4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |||
| 5 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | |||
| Sample 7 | Sample 7 | |||||||||||
| 1 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 3 | 3 | |||
| 2 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 4 | 4 | |||
| Digital Reactivity Grade | Manual Reactivity Grade | |||||||||||
| 3 | 4 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 3 | 3 | 3 | |
| 4 | 4 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 3 | 3 | 3 | |
| 5 | 4 | 4 | 4 | 4 | 4 | 3 | 3 | 3 | 3 | 3 | 3 | |
| Sample 8 | Sample 8 | |||||||||||
| 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 4 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 5 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| Sample 9 | Sample 9 | |||||||||||
| 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 4 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 5 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| Sample 10 | Sample 10 | |||||||||||
| 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 4 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| 5 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
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Summary:
Qualitative Agreement (manual reading):
| | | Expected
Results | Manual Reading | | | | | | | | | | | |
|--------|----|---------------------|----------------|----------|----------|----------|----------|----------|-------|-------|-------|-------|-------|-------|
| | | | Site1 | | Site 2 | | Site 3 | | | | | | | |
| | | | Reader 1 | Reader 2 | Reader 1 | Reader 2 | Reader 1 | Reader 2 | | | | | | |
| | | | % neg | % pos | % neg | % pos | % neg | % pos | % neg | % pos | % neg | % pos | % neg | % pos |
| Sample | n | pos/neg | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 1 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 2 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 3 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 4 | 30 | pos | 7% | 93% | 0% | 100% | 0% | 100% | 0% | 100% | 7% | 93% | 7% | 93% |
| 5 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 6 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 7 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 8 | 30 | neg | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% |
| 9 | 30 | neg | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% |
| 10 | 30 | neg | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% |
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| | | Expected
Results | Digital Reading | | | | | | | | | | | |
|--------|----|---------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|-------------------|
| | | | Site1 | | Site 2 | | Site 3 | | | | | | | |
| Sample | n | pos/neg | Reader 1
% neg | Reader 1
% pos | Reader 2
% neg | Reader 2
% pos | Reader 1
% neg | Reader 1
% pos | Reader 2
% neg | Reader 2
% pos | Reader 1
% neg | Reader 1
% pos | Reader 2
% neg | Reader 2
% pos |
| 1 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 2 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 3 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 4 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 5 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 6 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 7 | 30 | pos | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% |
| 8 | 30 | neg | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% |
| 9 | 30 | neg | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% |
| 10 | 30 | neg | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% | 100% | 0% |
Qualitative Agreement (digital reading):
Agreement between reader 1 and reader 2 at each site:
| % Overall Agreement (Positive/Negative) between operators (per site) | |||
|---|---|---|---|
| Site 1 | Site 2 | Site 3 | |
| Manual Reading | 99.7% | 100.0% | 100.0% |
| Digital Reading | 100.0% | 100.0% | 100.0% |
Reproducibility between lots
Lot to lot comparison study was performed on two reagent lots. Twenty clinically and/or analytically characterized samples were tested in duplicate.
The following comparisons were made:
- . NOVA view output: qualitative agreement.
- Digital image reading: qualitative agreement and grade agreement .
- . Manual image reading: qualitative agreement and grade agreement comparison.
Acceptance criteria:
- . Qualitative positive, negative and total agreement: ≥ 90%
- . Grade agreement: ≥ 90% within ± 1 reactivity grade
Qualitative Agreement
Qualitative positive agreements in two-way comparisons ranged from 91.7% to 100.0% Qualitative negative agreements in two-way comparisons ranged from 96.4% to 100.0% Qualitative total agreements in two-way comparisons ranged from 95.0%-100.0%
NOVA View Summary Table
| NOVA View | Negative agreement (%)
(95% CI) | Positive agreement (%)
(95% CI) | Total agreement (%)
(95% CI) |
|-------------------------|------------------------------------|------------------------------------|---------------------------------|
| Lot 046700 vs Lot RP003 | 96.4 (82.3-99.4) | 91.7 (64.6-98.5) | 95.0 (83.5-98.6) |
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Manual Summary Table
| NOVA View | Negative agreement (%)
(95% CI) | Positive agreement (%)
(95% CI) | Total agreement (%)
(95% CI) |
|-------------------------|------------------------------------|------------------------------------|---------------------------------|
| Lot 046700 vs Lot RP003 | 100.0 (86.2 – 100.0) | 100.0 (80.6– 100.0) | 100.0 (91.2 – 100.0) |
Digital Summary Table
| NOVA View | Negative agreement (%)
(95% CI) | Positive agreement (%)
(95% CI) | Total agreement (%)
(95% CI) |
|-------------------------|------------------------------------|------------------------------------|---------------------------------|
| Lot 046700 vs Lot RP003 | 100.0 (87.1-100.0) | 92.9 (68.5-98.7) | 97.5 (87.1-99.6) |
Grade agreement
All grades (100%) were within ±1 grade from each other for all samples in any pair-wise comparisons for manual and most grades were within ±1 grade from each other for all samples in any pair-wise comparison for digital reading, two comparisons were 100% and one comparison was 98%.
Manual – Agreement +/- 1 reactivity grade = 100%
| RP0003 | ||||||
|---|---|---|---|---|---|---|
| 046700 | 0 | 1 | 2 | 3 | 4 | Total |
| 0 | 24 | 0 | 0 | 0 | 0 | 24 |
| 1 | 0 | 9 | 0 | 0 | 0 | 9 |
| 2 | 0 | 0 | 1 | 0 | 0 | 1 |
| 3 | 0 | 0 | 0 | 4 | 0 | 4 |
| 4 | 0 | 0 | 0 | 0 | 2 | 2 |
| Total | 24 | 9 | 1 | 4 | 2 | 40 |
Digital - Agreement +/- 1 reactivity grade = 98%
| RP0003 | ||||||
|---|---|---|---|---|---|---|
| 046700 | 0 | 1 | 2 | 3 | 4 | Total |
| 0 | 26 | 0 | 0 | 0 | 0 | 26 |
| 1 | 1 | 3 | 0 | 0 | 0 | 4 |
| 2 | 0 | 0 | 2 | 0 | 0 | 2 |
| 3 | 0 | 0 | 0 | 0 | 0 | 0 |
| 4 | 0 | 0 | 1 | 1 | 6 | 8 |
| Total | 27 | 3 | 3 | 1 | 6 | 40 |
Linearity
The linearity study was performed by serially diluting 3 positive samples (one high positive, one medium positive and one low positive) from 1:10 to 1:5120. These samples were assed with the NOVA Lite DAPI dsDNA Crithidia luciliae Kit and read with the NOVA View, digital images were interpreted and confirmed. All slides were read with manual microscopy. Qualitative and semi-quantitative results (using a scale of 0 (negative) to 4 (strong positive)) were captured for the manual and digital reading.
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| Sample No. | Sample | NOVA View (SWT) | Manual | Digital |
|---|---|---|---|---|
| 1 | High Positive | >=320 | 640 | 640 |
| 2 | Medium Positive | 40 | 20 | 40 |
| 3 | Low Positive | 20 | 20 | 20 |
Three samples used in the study:
The sample dilutions and associated intensity grade results are summarized in the table below for manual microscopy:
| Sample No. | 10 | 20 | 40 | 80 | 160 | 320 | 640 | 1280 | 2560 | 5120 |
|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 4 | 4 | 4 | 4 | 3 | 2 | 1 | 0 | 0 | 0 |
| 2 | 2 | 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 3 | 1 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
The sample dilutions and associated intensity grades are summarized in the table below for digital reading:
| Sample No. | 10 | 20 | 40 | 80 | 160 | 320 | 640 | 1280 | 2560 | 5120 |
|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 4 | 4 | 4 | 4 | 3 | 2 | 1 | 0 | 0 | 0 |
| 2 | 2 | 2 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| 3 | 1 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
Interference
The interference study was performed according to CLSI EPO7-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition. A set of three specimens were tested (one negative, one positive, one strong positive) using the following Interfering substances (hemoglobin, bilirubin, triglycerides, cholesterol, Rituximab, Methylprednisolone, Cyclophosphamide, Methotrexate, Azathioprine, Ibuprofen, Naproxen, Hydroxychloroquine, Mycophenolate). All interferents were spiked into every specimen at three different concentrations in 10% of total specimen volume, and the resulting samples were assessed in triplicates with the NOVA Lite DAPI dsDNA Crithidia luciliae Kit. To assess interference with rheumatoid factor (RF), 10%, 30% and 50% (volume) RF positive sample was added to the test samples. All samples were processed with NOVA Lite DAPI dsDNA Crithidia luciliae kit and read with NOVA View. Digital images were interpreted and confirmed. Moreover, all slides were read by the same operator with manual microscopy. Acceptance criteria for the interference studies were grades obtained on samples with interfering substances are within ± 1 reactivity grade of those obtained on the control samples, spiked with diluent. No interference was detected with hemoglobin up to 200 mg/dL, bilirubin up to 100 mg/dL, triglycerides up to 1,000 mg/dL, cholesterol up to 224.3 mg/dL, rheumatoid factor up to 28.02 IU/mL, azathioprine up to 0.03 mg/ml, cyclophosphamide up to 4.1 mg/mL, hydroxychloroquine up to 0.224 mg/ml, ibuprofen up to 5 mg/mL, methotrexate up to 0.1 mg/mL, methy|prednisolone up to 0.85 mg/ml, mycophenolate up to 0.004 mg/ml, naproxen up to 5 mg/mL, rituximab up to 7.6 mg/mL, and belimumab up to 8 mg/mL.
14
Sample Stability and Handling
Three samples, encompassing negative, around the cut-off, and positive samples were tested in duplicates for up to 21 days while stored at 2-8°C, up to 48 hours while stored at room temperature, and after repeated freeze/thaw cycles up to 3 cycles. Results were compared to those obtained on control samples (day zero, stored at 2-8°C).
Acceptance criteria:
•NOVA View results of positive or negative do not change category (positive to negative or negative to positive) and are not different than the control sample.
•Manual reading reactivity grades are within ±1 grade of that of the control sample (stored at 2-8°C)
•Digital image interpretation reactivity grades are within ±1 grade of that of the control sample (stored at 2-8°C).
All samples fulfilled the acceptance criteria at each time point for each condition. Based on these results, we recommend that samples are stored up to 48 hours at room temperature, up to 7 days at 2-8°C and can be subjected to up to 3 freeze/thaw cycles (when samples are stored at or below -20°C).
Reagent Stability
Shelf life
To establish the initial claim for shelf life, accelerated stability studies were performed for up to 4 weeks at 37°C ± 3°C, where one week is equal to six months at 5 ± 3°C.
Accelerated stability testing was performed on all of the components of NOVA Lite DAPI dsDNA Crithidia luciliae Kit to support stability claim. The components within the kit are as follows: Slides, conjugate, Negative control, positive control, PBSII and Mounting Medium. Each week a new sealed kit was placed in the incubator, and all components were tested at the end of the experiment together with the one that was stored at 5 ± 3°C. The reactivity of the kits was calculated for each time point (compared to those obtained with 5 ± 3°C stored kit). All calculations were performed by comparing results of the kit stored at 5 ± 3°C (control) to those stored at 37 ± 3°C (test) for 1, 2, 3, and 4 weeks, where one week is equal to six months at 5 ± 3°C. Comparison of reactivity grades between incubated kits against the control kit within the same lot for manual and digital methods.
Acceptance criteria for two-year preliminary expiration dating: Reactivity grades of all samples/reagent controls run must be within ±1 reactivity grade of the control condition (week 0) for both manual and digital image interpretation for all three lots.
The acceptance criteria were successfully met with the accelerated lots tested. All samples tested against those run on the control kit (week 0) were within ±1 reactivity grade of the control kit.
In-use stability
Conjugate
Stability Claim: stable for 8 weeks after opening when stored at 2-8℃ given that they have not reached the expiration date found on the label.
15
During assessing open vial stability, conjugate bottle was opened and placed in the refrigerator. It was tested each weeks against a bottle that was left unopened each week using positive and negative controls on dsDNA Crithidia luciliae kits.
Conjugate is considered stable if the following criteria were met:
- Appearance: Clear liquid, free from foreign matter
- The grades from each reading are within ±1 grade from each other.
- . Fluorescence grading of >3+ for undiluted positive control and grading of 0 for undiluted negative control.
- Testing is comparable to control
The acceptance criteria were successfully met with all 8 weeks tested.
Controls
Stability Claim: stable for 8 weeks after opening when stored at 2-8℃ given that they have not reached the expiration date found on the label.
During assessing open vial stability, control bottles was opened and then closed in the refrigerator. It was tested each week for 8 weeks against a bottle that was left unopened each week using positive and negative controls on dsDNA Crithidia luciliae kits.
Conjugate is considered stable if the following criteria were met:
- Appearance: Clear liquid, free from foreign matter
- The grades from each reading are within ±1 grade from each other. ●
- . Fluorescence grading of >3+ for undiluted positive control and grading of 0 for undiluted negative control.
- Testing is comparable to control
The acceptance criteria were successfully met with all 8 weeks tested.
Real time stability
Real time stability testing has been scheduled to be performed each of the time points listed below on the NOVA Lite DAPI dsDNA Crithidia luciliae Kit, to verify the 2-year expiration that was assigned based on accelerated stability studies.
At the time of the submission, results were available up to 24 months for lot 1, 15 months for lot 2 and 19 months for lot 3. Complete studies will be done by January 2020.
Acceptance criteria: results should fall within their respective ranges.
All results to date were within the acceptance limits.
Cut-off, reference range
The recommended starting dilution, above which the result is reported as positive and below which the result is reported as negative, is 1:10. The manufacturer suggests performing two-fold dilutions and also recommends that each laboratory establish its own titering protocol.
16
Clinical performance characteristics
Clinical sensitivity, specificity
To assess clinical performance, a clinical study was performed by Inova Diagnostics on 766 clinically characterized serum samples. The distribution of the cohorts is in the table below:
| Diagnosis | N | NOVA View pos | Manual pos | Digital pos | |||
|---|---|---|---|---|---|---|---|
| Systemic Lupus Erythematosus (SLE) | 391 | 223 | 57% | 188 | 48% | 188 | 48% |
| Drug Induced Lupus | 20 | 1 | 5% | 1 | 5% | 1 | 5% |
| Infectious Disease | 60 | 8 | 13% | 9 | 15% | 8 | 13% |
| Vasculitis | 30 | 2 | 7% | 0 | 0% | 0 | 0% |
| Primary Antiphospholipid | |||||||
| Syndrome | 20 | 5 | 25% | 5 | 25% | 5 | 25% |
| Sjogren's Syndrome | 30 | 3 | 10% | 0 | 0% | 0 | 0% |
| Celiac Disease | 20 | 1 | 5% | 3 | 15% | 0 | 0% |
| Systemic Sclerosis | 30 | 2 | 7% | 3 | 10% | 3 | 10% |
| Idiopathic Inflammatory Myopathy | 20 | 4 | 20% | 3 | 15% | 2 | 10% |
| Mixed Connective Tissue Disease | 20 | 3 | 15% | 3 | 15% | 3 | 15% |
| Crohn's Disease | 20 | 1 | 5% | 1 | 5% | 1 | 5% |
| Grave's Disease | 20 | 0 | 0% | 0 | 0% | 0 | 0% |
| Hashimoto's Disease | 30 | 6 | 20% | 0 | 0% | 3 | 10% |
| Rheumatoid Arthritis | 35 | 2 | 6% | 2 | 6% | 2 | 6% |
| Autoimmune Hepatitis (AIH) | 20 | 4 | 20% | 3 | 15% | 1 | 5% |
| Total | 766 | 265 | 35% | 221 | 29% | 217 | 28% |
Sensitivity (on SLE) and specificity, calculated on the combined population, are shown below.
| N=766 | Sensitivity (%)
(95% CI) | Specificity (%)
(95% CI) |
|-----------|-----------------------------|-----------------------------|
| Manual | 48.1 (43.2-53.0) | 91.2 (87.9-93.7) |
| Digital | 48.1 (43.2-53.0) | 92.3 (89.1-94.6) |
| NOVA View | 57.0 (52.1-61.8) | 88.8 (85.2-91.6) |
Clinical studies 3 sites
Clinical study was completed at three sites using 269 (clinically characterized samples were tested on the NOVA Lite DAPI dsDNA (Crithidia luciliae) Kit according to its direction insert.
- a. Clinical Sensitivity/Specificity:
Sensitivity and specificity for all sites combined (269 x 3 sites= 807 total samples; 100 positive x 3 sites=300; 169 negative x 3 sites= 507)
| N=807 | Clinical Diagnosis | |
|---|---|---|
| Positive/per total SLE | Negative/per total non SLE | |
| NOVA View | 120/300 | 433/507 |
| Manual Reading | 98/300 | 484/507 |
| Digital Reading | 102/300 | 484/507 |
Correlation to clinical diagnosis for each mode at the three sites
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NOVA Lite® DAPI dsDNA Crithidia luciliae Kit
| Performance | NOVA View | Manual Reading | Digital Reading |
|---|---|---|---|
| Sensitivity % | |||
| (95% CI) | 120/300 | ||
| 40.0 (34.6-45.6) | 98/300 | ||
| 32.7 (27.6-38.2) | 102/300 | ||
| 34.0 (28.9-39.5) | |||
| Specificity % | |||
| (95% CI) | 433/507 | ||
| 85.4 (82.1-88.2) | 484/507 | ||
| 95.5 (93.3-97.0) | 484/507 | ||
| 95.5 (93.3-97.0) |
Summary: clinical overall agreement for all three sites
Correlation with SLE clinical diagnosis
| Sensitivity (95% CI)
| N=100 | NOVA View | Manual Reading | Digital Reading |
|---|---|---|---|
| Site 1 | 39/100 | 33/100 | 33/100 |
| 39.0 (30.0-48.8) | 33.0 (24.6-42.7) | 33.0 (24.6-42.7) | |
| Site 2 | 43/100 | 33/100 | 34/100 |
| 43.0 (33.7-52.8) | 33.0 (24.6-42.7) | 34.0 (25.5-43.7) | |
| Site 3 | 38/100 | 32/100 | 35/100 |
| 38.0 (29.1-47.8) | 32.0 (23.7-41.7) | 35.0 (26.4-44.7) |
Correlation with differential diagnosis
| Specificity (95% CI)
| N=169 | NOVA View | Manual Reading | Digital Reading |
|---|---|---|---|
| Site 1 | 143/169 | ||
| 84.6 (78.4-89.3) | 164/169 | ||
| 97.0 (93.3-98.7) | 165/169 | ||
| 97.6 (94.1-99.1) | |||
| Site 2 | 143/169 | ||
| 84.6 (78.4-89.3) | 159/169 | ||
| 94.1 (89.5-96.8) | 161/169 | ||
| 95.3 (90.9-97.6) | |||
| Site 3 | 147/169 | ||
| 87.0 (81.1-91.2) | 161/169 | ||
| 95.3 (90.9-97.6) | 158/169 | ||
| 93.5 (88.7-96.3) |
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NOVA Lite® DAPI dsDNA Crithidia luciliae Kit
| Site 1 | Site 2 | Site 3 | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Disease | N | NOVA View | Manual Reading | Digital Reading | NOVA View | Manual Reading | Digital Reading | NOVA View | Manual Reading | Digital Reading | |
| SLE | 100 | 39% | 33% | 33% | 43% | 33% | 34% | 38% | 32% | 35% | |
| Specificity Non-SLE | |||||||||||
| AIH | 20 | 80% | 95% | 95% | 60% | 85% | 85% | 80% | 90% | 85% | |
| APS | 20 | 85% | 100% | 100% | 95% | 100% | 100% | 90% | 100% | 100% | |
| AAV | 19 | 89% | 100% | 100% | 79% | 89% | 95% | 89% | 95% | 89% | |
| CD | 8 | 100% | 100% | 100% | 88% | 100% | 100% | 88% | 100% | 100% | |
| CKD | 2 | 100% | 100% | 100% | 0% | 100% | 100% | 100% | 100% | 100% | |
| COPD | 9 | 78% | 89% | 100% | 89% | 100% | 100% | 100% | 100% | 100% | |
| CrD | 6 | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | |
| HBV | 2 | 100% | 100% | 100% | 50% | 100% | 100% | 100% | 100% | 100% | |
| HCV | 5 | 100% | 100% | 100% | 80% | 100% | 100% | 100% | 100% | 100% | |
| HIV | 12 | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | |
| RA | 20 | 80% | 100% | 100% | 100% | 100% | 100% | 80% | 100% | 100% | |
| SjS | 20 | 90% | 100% | 100% | 100% | 100% | 100% | 85% | 100% | 100% | |
| SSc | 20 | 55% | 85% | 85% | 65% | 75% | 80% | 70% | 75% | 70% | |
| Syphilis | 6 | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% |
Summary: sensitivity/specificity, correlation with clinical diagnosis by disease
SLE= Systemic Lupus Erytematosus; AlH= Autoimmune Hepatitis; APS= Antiphospholipid Syndrome; AAV= ANCA Associated Vasculitis; CD= Celiac Disease; CKD= Chronic Kidney Disease; COPD= Chronic Obstructive Pulmonary Disease; CrD= Chrohns Disease; HBV= Hepatitis B; HCV= Hepatitis C; HIV= Human Immunodeficiency Virus; RA= Rheumatoid Arthritis; SjS= Sjogren's Syndrome; SSc= Systemic Sclerosis.
Expected values
Expected values were analyzed on 120 samples from apparently healthy subjects: 60 females, with mean age of 41 years (range of 18-73).
There were four (3.3%) positive results with manual interpretation and eleven (9.2%) positive results with NOVA View software interpretation and one (0.8%) positive results with digital interpretation with NOVA Lite DAPI dsDNA Crithidia luciliae Kit testing.
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Comparison with predicate device
Method comparison was performed with results obtained with the NOVA Lite DAPI dsDNA Crithidia luciliae Kit by NOVA View software interpretation and with the predicate device was performed using the same 744 serum samples (comprising of 391 serum samples from patients with SLE and 353 samples from patients with other diseases) tested in the clinical study (see table above).
Method comparison results are shown below:
| n=744 | Positive agreement (%)
95% CI) | Negative agreement (%)
95% CI) | Total agreement (%) |
|--------------------------------------|-----------------------------------|-----------------------------------|---------------------|
| 708205 manual vs
708215 manual | 87.8 (82.7-91.5) | 96.0 (94.0-97.4) | 93.7 |
| 708205 manual vs
708215 digital | 88.0 (82.9-91.7) | 94.6 (92.3-96.2) | 92.7 |
| 708205 manual vs
708215 NOVA View | 87.0 (81.8-90.9) | 85.3 (82.0-88.0) | 85.7 |
| 708205 Manual Grade | ||||||
|---|---|---|---|---|---|---|
| 708215 Manual | ||||||
| Grade | 0 | 1 | 2 | 3 | 4 | Total |
| 0 | 510 | 24 | 1 | 0 | 1 | 536 |
| 1 | 19 | 34 | 22 | 1 | 0 | 76 |
| 2 | 2 | 13 | 34 | 4 | 0 | 53 |
| 3 | 0 | 1 | 6 | 21 | 6 | 34 |
| 4 | 0 | 2 | 1 | 8 | 34 | 45 |
| Total | 531 | 74 | 64 | 34 | 41 | 744 |
+/- 2 reactivity grade
99.6%
| 708205 Manual Grade | ||||||
|---|---|---|---|---|---|---|
| 708215 Digital Grade | 0 | 1 | 2 | 3 | 4 | Total |
| 0 | 507 | 19 | 5 | 4 | 1 | 536 |
| 1 | 19 | 15 | 26 | 10 | 6 | 76 |
| 2 | 5 | 3 | 12 | 24 | 9 | 53 |
| 3 | 0 | 1 | 1 | 4 | 28 | 34 |
| 4 | 1 | 0 | 1 | 1 | 42 | 45 |
| Total | 532 | 38 | 45 | 43 | 86 | 744 |
+/- 2 reactivity grade
98.4%
System description
NOVA View Automated Fluorescence Microscope and Software
NOVA View Automated Fluorescence Microscope
NOVA View® Automated Fluorescence Microscope is an automated system consisting of a fluorescence microscope and software that acquires, analyzes, stores and displays digital images of stained indirect immunofluorescent slides. It is intended as an aid in the detection and classification of certain antibodies
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by indirect immunofluorescence technology. The device can only be used with cleared or approved in vitro diagnostic assays that are indicated for use with the device. A trained operator must confirm results generated with the device.
The NOVA View device has been cleared by the FDA in DEN140039. Subsequently, the use of NOVA View with ANCA modules has been cleared in K161258. Device description and Principle of Operation were described in DEN140039 and K161258 remained unchanged for this submission.
Software
Level of Concern: Moderate.
The NOVA View software version in this submission contains the following changes compared to the version in K161258:
Addition of CLIFT module and CLIFT SWT application.
When CLIFT slides are analyzed by NOVA View, digital images of representative fields of view of the well are captured. At the same time when digital images are taken, NOVA View measures the FITC light intensity of the cells that are included in the region. NOVA View reports the measured fluorescence intensity in units of Light Intensity Units (LIU).
NOVA View provides the trained operator with the acquired digital images and the following supportive information:
-
LIU value
-
Negative/positive classification
NOVA View CLIFT Single Well Titer (SWT) This assay is compatible with NOVA View SWT.
The SWT is a software application that estimates the endpoint titer (e.g., the highest dilution that gives positive result) for wells with a positive reaction with CLIFT, based on the obtained fluorescence intensity. The highest titer that can be differentiated is 1:320. Above this value the NOVA View reports > 320 titer. SWT validation was part of the between sites reproducibility study.
The SWT function was established using 22 dsDNA positive samples that represent various levels of antibodies. Two-fold serial dilutions were made for each sample, starting from 1:10, up to 1:40960. Each dilution was processed on dsDNA Crithidia luciliae slides, and the results were interpreted by NOVA View, digital and manual reading. Results were used to establish the intensity curves based on 4-PL logistic curve fitting. NOVA View uses these built-in curves for the determination of the titer
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The validation of the SWT function was performed using 31 positive samples were serially diluted and read using NOVA View and manual microscope (manual end-point titer). SWT was determined for all samples.
Acceptance criteria:
- . SWT is within ± 2 dilution steps of that of the manual end-point titer and the digital titer.
Based on 31 samples, 80.6% of SWT results were within ± 1 dilution step of that of the manual titer, and 83.9% were within ±1 dilution step of that of the digital titer, and 93.3% of SWT results were within ± 2 dilution steps of that of the manual titer and 93.5% were within ± 2 dilution steps of that of the digital titer. 2 out of the 31 samples were outside of this range.
| SWT, n=31 | within ± 1 dilution step | within ± 2 dilution steps |
|---|---|---|
| Manual end-point | 80.6% | 93.5% |
| Digital end-point | 83.9% | 93.5% |
Additionally, SWT validation was part of the between sites reproducibility study.
Seven positive samples were assayed by all three testing sites (including Inova). 100% (14 out of 14) of SWT results at the two external sites were within ± 1 dilution step of that of the manual titer, and 92.9% (13 out of 14) were within ± 1 dilution step of that of the digital titer; 100% of SWT results were within ± 2 dilution steps of that of both the manual titer and digital titer.