AESKUSLIDES ANCA Ethanol, AESKUSLIDES ANCA Formalin

{0}------------------------------------------------ # Please wait... If this message is not eventually replaced by the proper contents of the document, your PDF viewer may not be able to display this type of document. You can upgrade to the latest version of Adobe Reader for Windows®, Mac, or Linux® by visiting http://www.adobe.com/go/reader_download. For more assistance with Adobe Reader visit http://www.adobe.com/go/acrreader. Windows is either a registered trademark of Microsoft Corporation in the United States and/or other countries. Mac is a trademark of Apple Inc., registered in the United States and other countries. Linux is the registered trademark of Linus Torvalds in the U.S. and other countries. {1}------------------------------------------------ Image /page/1/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: a symbol on the left and the text "FDA U.S. FOOD & DRUG ADMINISTRATION" on the right. The symbol on the left is a stylized representation of a human figure, while the text on the right is written in a bold, sans-serif font. The word "FDA" is in a blue box. March 23, 2018 Aesku.Diagnostics GmbH & Co. KG Sandra Reuter Regulatory Affairs Manager Mikro-Forum-Ring 2 Wendelsheim, Rheinland-Pfalz55234 Germany Re: K172461 Trade/Device Name: AESKUSLIDES® ANCA Ethanol AESKUSLIDES® ANCA Formalin Regulation Number: 21 CFR 866.5660 Regulation Name: Multiple autoantibodies immunological test system Regulatory Class: Class II Product Code: MOB Dated: July 31, 2017 Received: August 14, 2017 Dear Sandra Reuter: We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration. listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading. If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal {2}------------------------------------------------ statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance. For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100). Sincerely. Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health Enclosure {3}------------------------------------------------ Image /page/3/Picture/0 description: The image is a logo for AESKU.DIAGNOSTICS. The logo features a green square with a white symbol inside, followed by the text "AESKU.DIAGNOSTICS" in gray. Below the text, there is a tagline that reads "THE DIAGNOSTIC TOOL THAT WORKS" in a smaller, lighter gray font. ## 510(k) Summary - A. 510(k) Number: K172461 - B. Purpose for Submission: New Device ## C. Measurand: Anti-neutrophil cytoplasmic autoantibodies (ANCA) ## D. Type of Test: Qualitative and semi-quantitative, indirect immunofluorescence ## E. Applicant: AESKU.Diagnostics GmbH & Co.KG Mikro-Forum-Ring 2 55234 Wendelsheim Germany Contact Person: Dr. Sandra Reuter Regulatory Affairs registrations@aesku.com p: +49 6734 9622 0 prepared on March 23rd, 2018 #### F. Proprietary and Established Names: AESKUSLIDES® ANCA Ethanol AESKUSLIDES® ANCA Formalin ## G. Regulatory Information: 1. Regulation section: §CFR 866.5660 - Multiple autoantibodies immunological test system 2. Classification: Class II 3. Product code: MOB - Anti-neutrophil cytoplasmic antibodies (ANCA) 4. Clinical use Immunology (82) {4}------------------------------------------------ Image /page/4/Picture/1 description: The image shows the logo for AESKU.DIAGNOSTICS. The logo consists of a green square with a white symbol inside, followed by the text "AESKU.DIAGNOSTICS" in gray. Below the text, there is a tagline that reads "THE DIAGNOSTIC TOOL THAT WORKS" in a smaller, lighter gray font. #### H. Intended Use: - 1. Intended use(s): AESKUSLIDES® ANCA is an indirect immunofluorescence assay utilizing human neutrophil granulocyte coated slides, fixed with Ethanol or Formalin, as a substrate for the qualitative and semi-quantitative determination of anti-neutrophil cytoplasmic autoantibodies (ANCA) in human serum by manual microscopy or with the HELIOS® AUTOMATED IFA SYSTEM. This in vitro diagnostic assay is used as an aid for the diagnosis of ANCA-associated vasculitides (AAV) in conjunction with other clinical and laboratory findings. All suggested results obtained with the HELIOS AUTOMATED IFA SYSTEM must be confirmed by trained personnel. - 2. Indication(s) for use: Same as intended use - 3. Special conditions for use statement(s): - 1. For prescription use only 2. This device is only for use with reagents that are indicated for use with the device. 3. The device is for use by a trained operator in a clinical laboratory setting. 4. All software-aided results must be confirmed by the trained operator. 5.For use only by manual microscopy or with HELIOS® AUTOMATED IFA SYSTEM. #### l. Device Description: AESKUSLIDES ANCA is an indirect immunofluorescence assay utilizing human neutrophil granulocyte coated slides, fixed with Ethanol or Formalin, as a substrate for the qualitative and semi-quantitative determination of anti-neutrophil cytoplasmic autoantibodies (ANCA) in human serum by manual microscopy or with the HELIOS® AUTOMATED IFA SYSTEM. This in vitro diagnostic assay is used as an aid for the diagnosis of ANCA-associated vasculitides (AAV) in coniunction with other clinical and laboratory findings. All suggested results obtained with the HELIOS AUTOMATED IFA SYSTEM must be confirmed by trained personnel. Slides coated with human neutrophil granulocytes for autoantibody detection are fixated by two different methods: ethanol (EtOH) fixation or formalin fixation. Ethanol fixation allows cell components to move through the cells after the fixation process. Formalin fixation causes cellular components to cross-link (a movement of cellular components is abrogated and the patterns are distinct). By processing serum on both Ethanol and Formalin-fixed slides, the user can confirm if the pattern is C-, P-, or A-ANCA, according to the table below. {5}------------------------------------------------ Image /page/5/Picture/0 description: The image shows the logo for AESKU.DIAGNOSTICS. The logo consists of a green square with a white symbol inside, followed by the text "AESKU.DIAGNOSTICS" in gray. Below the text, there is a tagline that reads "THE DIAGNOSTIC TOOL THAT WORKS" in a lighter shade of gray. | Ethanol-fixed Result | Formalin-Fixed Result | Pattern | |-------------------------|--------------------------|------------------| | Cytoplasmic (C-ANCA) | Cytoplasmic (C-ANCA) | C-ANCA | | Perinuclear (P-ANCA) | Cytoplasmic (C-ANCA) | P-ANCA | | Perinuclear (P-ANCA) | Negative / unclear | Confirm with ANA | | Mixed (P-ANCA + C-ANCA) | Negative / weak positive | A-ANCA | | Very Perinuclear | A-ANCA | A-ANCA | ANCA Formalin test is not intended to be used by itself, but in conjunction with ANCA Ethanol test. The HELIOS Vasculitis Pattern Plus software is able to detect both C- and P-ANCA. A-ANCA are reported as undefined positive. All suggested results obtained with the HELIOS AUTOMATED IFA SYSTEM must be confirmed by trained personnel. Each kit of ANCA Ethanol and ANCA Formalin contains (Quantity depends on product variant): - Slides, each containing 6 or 12 wells coated with human neutrophils (ethanol fixation)/ human neutrophils (formalin fixation) cells - -2.0 ml/ 4.0 ml vial containing Fluorescein (FITC) labelled Anti-human Antibody IgG conjugate in a solution of BSA, ready for use - -0.5 ml vial of positive control containing human serum (diluted), ready for use - -0.5 ml vial of negative control containing diluted human serum, ready for use - 8.0 ml vial of mounting medium containing a solution of glycerol and PBS, ready for use - 70 ml bottle of sample buffer, containing BSA, PBS and ready for use - - -100 ml bottle of wash buffer, concentrated buffer 1:10 in distilled water, containing PBS | Standard Ref. | Description | Tests | |------------------|--------------------------|-------| | 54.100.US | ANCA Ethanol (12 wells) | 120 | | 54.101.US | ANCA Formalin (12 wells) | 120 | | 54.050.US | ANCA Ethanol (6 wells) | 60 | | 54.051.US | ANCA Formalin (6 wells) | 60 | | 54.100.US.Demo | ANCA Ethanol (12 wells) | 24 | | 54.100.US.Bulk5 | ANCA Ethanol (12 wells) | 600 | | 54.100.US.Bulk10 | ANCA Ethanol (12 wells) | 1200 | | 54.101.US.Demo | ANCA Formalin (12 wells) | 24 | | 54.101.US.Bulk5 | ANCA Formalin (12 wells) | 600 | | 54.101.US.Bulk10 | ANCA Formalin (12 wells) | 1200 | {6}------------------------------------------------ ## Not provided in the kit: | Ref. | Reagent | | Quantity<br>/ Volume | Description | Ready<br>to use | |-------|-----------------------|----|----------------------|--------------------------------------------------------------------------------------------------------------------------------|-----------------| | EBIFA | Evans<br>Blue<br>0.2% | 1x | 3ml | Capped white: Blue coloured<br>solution<br>Containing: PBS, Evans<br>Blue.<br>Dilute the Evans Blue 0.2%<br>1:3000 in 1x WBIFA | NO | ## HELIOS AUTOMATED IFA SYSTEM (k153117) or equivalent manual microscope. ## J. Substantial Equivalence Information: - 1. Predicate device name(s): NOVA Lite ANCA - 1. Predicate 510(k) number(s): K961340 - 2. Comparison with predicate device: #### Table 7: comparison table with predicate device | | Item | Predicate<br>NOVA Lite ANCA | AESKUSLIDES® ANCA | |--------------|------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Similarities | Intended Use | NOVA Lite® ANCA is an indirect<br>immunofluorescent assay for the<br>screening and semi-quantitative<br>determination of anti-neutrophil<br>cytoplasmic antibodies (ANCA) in<br>human serum. The presence of<br>antineutrophil cytoplasmic<br>antibodies can be used in<br>conjunction with other serological<br>tests and clinical findings aids in<br>the assessment of various<br>systemic vasculitides. | AESKUSLIDES ANCA is an indirect<br>immunofluorescence assay utilizing human<br>neutrophil granulocyte coated slides, fixed with<br>Ethanol or Formalin, as a substrate for the<br>qualitative and semi-quantitative determination of<br>anti-neutrophil cytoplasmic autoantibodies (ANCA)<br>in human serum by manual microscopy or with the<br>HELIOS® AUTOMATED IFA SYSTEM.<br>This in vitro diagnostic assay is used as an aid for<br>the diagnosis of ANCA-associated vasculitides<br>(AAV) in conjunction with other clinical and<br>laboratory findings.<br>All suggested results obtained with the HELIOS<br>AUTOMATED IFA SYSTEM must be<br>confirmed by trained personnel. | | | Methology | Immunofluorescence assay (IFA) | same as Predicate | | | Procedure | Standard IFA technique<br>qualitative and semi-quantitative<br>titer | same as Predicate | | | Results | Serum | same as Predicate | | | Samples Matrix | anti-neutrophil cytoplasmic<br>autoantibodies (ANCA) | same as Predicate | | | Analyte | ethanol-fixed human neutrophil /<br>formalin-fixed human neutrophil | same as Predicate | | | Antigen | FITC | same as Predicate | | | Fluorescence<br>Marker | cANCA Positive, pANCA Positive<br>and one negative control | same as Predicate | | | Controls | Anti-Human IgG Conjugate | same as Predicate | | | conjugate | 1:20 | same as Predicate | | | Screening<br>dilution | 2 - 8 °C | same as Predicate | | | Storage | | | {7}------------------------------------------------ Image /page/7/Picture/0 description: The image shows the logo for AESKU.DIAGNOSTICS. The logo features a green square with a white symbol inside. To the right of the square, the text "AESKU.DIAGNOSTICS" is written in gray. Below the text, the phrase "THE DIAGNOSTIC TOOL THAT WORKS" is written in a smaller, lighter gray font. | | Slides | 6 or 12 wells coated with antigen | same as Predicate | |-------------|-------------------------------------------|-----------------------------------|-----------------------------------------------------------------------------------| | Differences | Shelf life | 18 months | 24 months for ANCA Ethanol<br>18 months for ANCA Formalin | | | Manual<br>Interpretation of<br>result | Manual fluorescence microscopy | Manual fluorescence microscopy or with HELIOS<br>w/ trained operator verification | | | Automated<br>interpretation of<br>results | N/A | HELIOS w/ trained operator verification | ## K. Standard/ Guidance Document Referenced (if applicable): Table 8: List of Standards / Guidance Documents | # | Standards Title | Version | |----|--------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------| | 1 | ISO 14971 - Medical Devices - Application of risk management to medical<br>devices | Second<br>Edition | | 2 | IEC 62366 - Medical Devices - Part 1: Application Of Usability Engineering To<br>Medical Devices [Including CORRIGENDUM 1 (2016)] | Edition 1.0 | | 3 | 15223-1 - Medical Devices - Symbols To Be Used With Medical Device Labels,<br>Labelling, and Information to be supplied - Part 1: General Requirements | Second<br>Edition | | 4 | Interference Testing in Clinical Chemistry | EP07-A2 | | 5 | Evaluation of Detection Capability for Clinical Laboratory Measurement<br>Procedures | EP17-A2 | | 6 | Evaluation of Stability of In Vitro Diagnostic Reagents | EP25-A | | 7 | Defining, Establishing and Verifying Reference Intervals in the Clinical<br>Laboratory | EP28-A3c | | 8 | Evaluation of the Linearity of Quantitative Measurement Procedures A<br>Statistical Approach | EP06-A | | 9 | Procedures for the Handling and Processing of Blood Specimens for Common<br>Laboratory Tests; Approved Guideline-Fourth Edition | GP44-A4 | | 10 | Guidance for Industry and FDA Staff: Recommendations for Anti-Nuclear<br>Antibody (ANA) Test System Premarket (510k) Submissions | - | | 11 | Factors to Consider Regarding Benefit-Risk in Medical Device Product<br>Availability, Compliance, and Enforcement Decisions | - | | 12 | Applying Human Factors and Usability Engineering to Medical Devices | - | ## L. Test Principle AESKUSLIDES® ANCA is an indirect fluorescent antibody assay utilizing human neutrophil granulocyte coated slides, fixed with Ethanol or Formalin, as a substrate for the qualitative and semi-quantitative determination of anti-neutrophil cytoplasmic autoantibodies (ANCA) in human serum by manual microscopy or with HELIOS® AUTOMATED IFA SYSTEM. This in vitro diagnostic assay is used as an aid for the diagnosis of ANCA-associated vascultitides (AAV) in conjunction with other clinical and laboratory findings. All suggested results obtained with the HELIOS® AUTOMATED IFA SYSTEM must be confirmed by trained personnel. Manual interpretation of AESKUSLIDES® ANCA Ethanol: The two main patterns seen on an ethanol-fixed substrate are perinuclear (P-ANCA) and cytoplasmic (C-ANCA): {8}------------------------------------------------ Image /page/8/Picture/1 description: The image is a logo for AESKU.DIAGNOSTICS. The logo features a green square with a white image of a medical symbol on the left. To the right of the square is the text "AESKU.DIAGNOSTICS" in a gray sans-serif font. Below the text is the tagline "THE DIAGNOSTIC TOOL THAT WORKS" in a smaller, lighter gray font. - -C-ANCA presents as coarse speckled cytoplasmic fluorescence, often with accentuated staining between the nuclear lobes. This pattern is characteristic for antibodies reacting with PR3. - -P-ANCA presents as perinuclear staining with or without nuclear extension. This pattern is usually characteristic for antibodies reacting with MPO. Note that anti-nuclear antibody (ANA) positive samples (containing anti-DNA/histones) may react with the nuclei of ethanol-fixed neutrophils, causing nuclear staining, and may mask or mimic the P-ANCA pattern(s). A third pattern, less commonly seen, is called atypical ANCA (A-ANCA or X-ANCA): - । A-ANCA presents as a cytoplasmic and perinuclear or very perinuclear staining on ethanol-fixed neutrophil substrate and usually becomes negative on formalin fixed substrate. ## Manual interpretation of AESKUSLIDES® ANCA Formalin: On formalin fixed substrate, both MPO and PR3 antibodies appear as coarse cytoplasmic granular staining with interlobular accentuation. Results from both AESKUSLIDES® ANCA Ethanol and AESKUSLIDES® ANCA Formalin provide further information on the antibodies present in the serum. The fluorescence intensity level is the intensity of the specific fluorescence expressed as a numeric value. These values, if present, are reported as a number between "0" (no specific fluorescence) and "4+" (very strong visible reaction). | Intensity | Interpretation | |-----------|---------------------------------------------------------------------------------------------| | 4+ | high positive<br>maximal fluorescence, very strong visible reaction; brilliant yellow-green | | 3+ | positive<br>strong visible reaction; less brilliant as 4+; yellow-green fluorescence | | 2+ | positive<br>moderate visible reaction; definite but dull yellow-green fluorescence | | 1+ | positive<br>weak visible reaction, very dim subdued fluorescence | | 0 | negative<br>no specific fluorescence | AESKU recommends a screening dilution of 1:20, followed by serial dilutions for semiquantitative determinations and suggests each laboratory establish its own screening dilution and titration scheme based on its population and instrumentation. #### Qualitative evaluation A serum dilution is considered negative for ANCA antibodies if the cells exhibit < 1+ fluorescence of the cytoplasm or nucleus. Likewise, a serum dilution is considered positive for ANCA antibodies if the cells exhibit ≥ 1+ fluorescence of the cytoplasm or nucleus. A sample is considered positive for ANCA antibodies if it exhibits ≥ 1+ fluorescence of the cytoplasm or nucleus at a sample dilution of 1:20 or greater. Operators should report all titers and specific fluorescence staining seen. {9}------------------------------------------------ Image /page/9/Picture/0 description: The image is a logo for AESKU.DIAGNOSTICS. The logo features a green square with a white image inside. The image appears to be a stylized representation of a chromosome. To the right of the square is the text "AESKU.DIAGNOSTICS" in gray, with the word "AESKU" being larger and bolder than "DIAGNOSTICS". Below the company name is the tagline "THE DIAGNOSTIC TOOL THAT WORKS" in a smaller, lighter gray font. ## Semi-quantitative evaluation The endpoint titer is defined as the highest sample dilution factor for which specific fluorescence of the cytoplasm or nucleus is identifiable. The titers are classified as: - 1:20 and 1:40 are considered low titers - 1:80 and 1:160 are considered medium titers - 1:320 and greater are considered high titers ## Automated instrument interpretation of test results #### Software After slides are processed by the HELIOS®, digital images of representative fields of view in the well are captured and stored on the computer system. The HELIOS Pattern Recognition software recognizes the pattern of the captured image by using SVM (Support Vector Machine) technology. After image pre-processing, feature extraction and classification, the software delivers the results. All suggested results given by the HELIOS® software must be confirmed within the Result Confirmation tool by a trained operator. The HELIOS PATTERN RECOGNITON software tool identifies the following immunofluorescence patterns, abbreviated below: | Abbreviation | Pattern | |--------------|------------------------------| | P-ANCA | perinuclear | | C-ANCA | cytoplasmic | | unk | unknown (undefined positive) | | neg | negative | The HELIOS DEVICE SOFTWARE examines the fluorescence intensity and uses an analysis algorithm which takes exposure and pixel frequency into account. It examines relevant regions of the captured images, such as the fluorescence cell area and subtracts the background, to provide an assessment of positive or negative results. | Intensity | Interpretation | |-----------|----------------| | + | positive | | - | negative | Trained operators must confirm all suggestions. Intensity and End Point Titers are identified by the HELIOS DEVICE SOFTWARE and pattern suggestions are made by the HELIOS PATTERN RECOGNITION tool. AESKU recommends a screening dilution of 1:20, followed by serial dilutions for semiquantitative determinations and suggests each laboratory establish its own screening dilution and titration scheme based on its population {10}------------------------------------------------ #### M. Performance Characteristics: ## 1. Analytical performance During the study 4 different methods have been applied: | Method | Processing | Imaging | Reading/Evaluation od<br>Slide | Alternate Name of<br>Method | |--------|------------|-----------|----------------------------------------|--------------------------------------| | A | Automated | Automated | Automated (Software<br>Interpretation) | HELIOS | | B | Automated | Automated | Manual (read of digital<br>image) | HELIOS User<br>Evaluation | | C | Manual | Manual | Manual (read of<br>microscope field) | Manual<br>AESKUSLIDES<br>ANCA | | D | Manual | Manual | Manual (read of<br>microscope field) | Manual - Predicate<br>NOVA Lite ANCA | Method D is the predicate for method C In the clinical study the method comparison was: Method D vs Method C Method C vs. Method A, B Method B vs Method A Nomenclature and acronyms used in studies Table 9: Nomenclature and acronyms | Nomenclature | Description/ Also known as | |------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------| | AESKUSLIDES® ANCA<br>HELIOS® AUTOMATED<br>IFA SYSTEM | ANCA<br>HELIOS | | Manual or Manual Reading | Results obtained by the operator after<br>reading and interpreting the slides with a<br>traditional fluorescence microscope | | Method A | HELIOS + AESKUSLIDES ANCA<br>w/slides read by HELIOS | | Method B | HELIOS + AESKUSLIDES ANCA<br>w/slides read by User | | Method C | Manual AESKUSLIDES ANCA w/slides<br>read by User | | Method D | Manual Nova Lite ANCA w/slides read<br>by User | | P-ANCA | perinuclear | | C-ANCA | cytoplasmic | | unk | unknown (undefined positive) | | ANCA | anti-neutrophil cytoplasmic<br>autoantibodies | | Negative IFA score/result,<br>negative | neg, negative | | Positive IFA score/result | pos, positive | | Diseases Abbreviation | Diseases Name | | SLE | Systemic lupus erythematosus | | MPA | Microscopic polyangiitis | | UC | Ulcerative colitis | {11}------------------------------------------------ Image /page/11/Picture/0 description: The image contains the logo for AESKU.DIAGNOSTICS. The logo features a green square with a white stylized image of a medical device. To the right of the square, the text "AESKU.DIAGNOSTICS" is displayed in gray, with "AESKU" in a larger, bolder font than "DIAGNOSTICS". Below the company name, the tagline "THE DIAGNOSTIC TOOL THAT WORKS" is written in a smaller, lighter gray font. | PMR | Polymyalgia rheumatica | |-----|------------------------------| | SpA | Spondyloarthropathy | | RA | Rheumatoid Arthritis | | AAV | ANCA-associated vasculitides | | HCV | Hepatitis C Virus | | HBV | Hepatitis B Virus | ## Serum sample: 630 Clinical samples have been used for the Clinical Evaluation and Method Comparison studies described herein. An exhaustive search for US Clinical Samples from 10 BioBanks (BioChain, BioReclamationIVT, Bioserve, ConversantBio, Cureline, DiscoveryLifeSciences, iSpecimen, Precision for Medicine, ProMedDx, and Vitrologic) has been performed and 510 Clinical Samples have been found. The samples have been selected according to their diagnosis, to reflect all important diagnosis for this study (see table). The sample set has been completed with 120 serum samples from a German University Hospital to complement some rare, but important diagnosis (70 sera from patients with Wegener's Granulomatosis, 25 sera from patients with MPA and 25 sera from patients with Churg-Strauss Syndrome). | Study Sample Set | | | | |------------------------------------------------|-------------------------------------------|------------------------------------------------------|-----| | | Sample Definition | Diagnosis | n | | Target<br>Diagnosis | ANCA<br>associated<br>Vasculitis<br>(AAV) | Wegener's Granulomatosis | 80* | | | | Microscopic polyangiitis (MPA) | 29* | | | | Churg-Strauss Syndrome | 26* | | Characterized<br>MPO/PR3/ANCA<br>positive sera | | PR3 positive | 25 | | | | MPO positive | 25 | | | | P-ANCA positive | 50 | | | | C-ANCA positive | 20 | | | | | | | Control<br>diagnosis | Autoimmune<br>Liver<br>Diseases | Autoimmune Hepatitis (AIH) | 8 | | | | Autoimmune Hepatitis/ Primary biliary<br>cholangitis | 6 | | | | Primary biliary cholangitis (PBC) | 11 | | | | Primary sclerosing cholangitis (PSC) | 10 | | Control<br>diagnosis | Inflammatory<br>bowel<br>diseases | Ulcerative Colitis | 71 | | | | Crohn's disease | 40 | | | | Inflammatory bowel disease (IBD) | 9 | | | Other<br>rheumatic<br>diseases | Rheumatoid Arthritis (RA) | 12 | | | | Systemic Lupus Erythematosus (SLE) | 30 | | | | Scleroderma | 19 | | | | Myositis | 1 | | | Infections | Hepatitis C Virus (HCV ) | 16 | | | | Hepatitis B Virus (HBV ) | 8 | | | | Polymyalgia rheumatica (PMR) | 18 | Table 10: Serum samples set for clinical and method comparison studies {12}------------------------------------------------ Image /page/12/Picture/0 description: The image contains the logo for AESKU.DIAGNOSTICS. The logo features a green square with a white image inside, resembling a stylized medical symbol. To the right of the square, the text "AESKU.DIAGNOSTICS" is displayed in a gray, sans-serif font. Below the company name, the tagline "THE DIAGNOSTIC TOOL THAT WORKS" is written in a smaller, lighter gray font. | | Vasculitides<br>NOT<br>associated<br>with ANCA | Giant cell artertitis (GCA) | 4 | |-----|------------------------------------------------|-----------------------------|-----| | | Purpura | | 1 | | | Leukemia | Lymphoma, Myeloma | 15 | | | Other<br>diagnosis | Celiac disease | 10 | | | | Chronic kidney disease | 32 | | | | Sinusitis | 22 | | | | Asthma | 32 | | Sum | | | 630 | *samples from Germany: 70 sera from patients with Wegener's Granulomatosis, 25 sera from patients with MPA and 25 sera from patients with Churg-Strauss Syndrome The diagnosis criteria of the different samples have been made in agreement with diagnostic standards used in the U.S and Germany. A written statement from different serum suppliers is available on request. Standard criteria are for example ACR criteria. The US sample set was also selected to contain different ethnic groups (e.g. White, Black/Black African, Asian, Hispanic) to reflect the ethnic composition of the US population as good as possible. The samples have been checked for purity and volume by visual inspection and for further contaminations by an in house pre-screening before inclusion into the study. All 630 sera passed and were determined suitable for the study. All serum samples which have been used for this studies have been shipped and stored at -20°C. Upon arrival from the Biobanks they were thawed once to aliquot them for the different studies. Subsequently, samples were frozen again. Therefore, samples underwent 2-3 freezing-thawing cycles before they were used in the studies. To confirm that repeated thawing and freezing and also long term storage at -20°C has no influence on the samples we have performed the following two studies: All serum samples which have been used for this studies have been shipped and stored at -20°C. Upon arrival from the Biobanks they were thawed once to aliquot them for the different studies. Subsequently, samples were frozen again. Therefore, samples underwent 2-3 freezing-thawing cycles before they were used in the studies. To confirm that repeated thawing and freezing and also long term storage at -20°C has no influence on the samples we have performed the following two studies: ## Serum Stability AESKUSLIDES® ANCA ## Procedure: Serum Stability was shown by testing 10 serum samples (7 positive, 3 negative). For each serum, one aliquot has been prepared that serves as control (no freezethawing). A second aliquot for each serum has been prepared that underwent 4 freeze-thaw cycles. Then, all samples and controls have been analyzed on AESKUSLIDES® ANCA Ethanol and ANCA Formalin according to the IFUs. Slides were read by two independent readers. Each freeze/thaw sample was compared to the respective control sample. {13}------------------------------------------------ | Table 11: Serum sample set for serum stability | | | | AESKUSLIDES ANCA Formalin | | | |------------------------------------------------|---------|----------------------------|-----------------|---------------------------|-----------------------------|-----------------| | Sample<br>ID | Pos/Neg | Pattern<br>ANCA<br>Ethanol | Grading | Pos/Neg | Pattern<br>ANCA<br>Formalin | Grading | | 1 | Pos | P-ANCA | medium positive | Pos | C-ANCA | medium positive | | 2 | Pos | P-ANCA | high positive | Pos | C-ANCA | high positive | | 3 | Pos | P-ANCA | high positive | Pos | C-ANCA | high positive | | 4 | Pos | C-ANCA | low positive | Pos | C-ANCA | medium positive | | 5 | Pos | C-ANCA | high positive | Pos | C-ANCA | high positive | | 6 | Pos | C-ANCA | medium positive | Pos | C-ANCA | medium positive | | 7 | Pos | A-ANCA | medium positive | Neg | Neg | negative | | 8 | Neg | Neg | negative | Neg | Neg | negative | | 9 | Neg | Neg | negative | Neg | Neg | negative | | 10 | Neg | Neg | negative | Neg | Neg | negative | #### Acceptance criteria: - Pos/Neg/Overall Agreement: All positive sera have to be found positive and all negative sera have to be found negative. Overall Agreement: > 85%. - -All patterns have to be found correctly. - -Pattern Agreement: > 85% - -FI is allowed to differ maximum + or - 1 from the expected value. FI Agreement: > 85%. #### Results All samples fulfilled the above criteria. No differences were observed between sera thawed a single time and sera that had undergone 4 cycles of freeze/thawing: - -Positive/Negative/Overall Agreement was 100% for both AESKUSLIDES® ANCA Ethanol and Formalin. All positive samples have been found positive, all negative samples have been found negative. - -Pattern Agreement was 100% for both AESKUSLIDES® ANCA Ethanol and Formalin. All patterns have been found as expected. - FI agreement was 100% for both AESKUSLIDES® ANCA Ethanol and Formalin. -No deviations of fluorescence intensities greater than +/-1 from the initial value have been observed. #### Conclusions All criteria are fulfilled in each test. The results show that multiple freeze-thaw cycles have no effect on test results. {14}------------------------------------------------ ## Long Term Serum Stability AESKUSLIDES® ANCA To show that long term storage of serum samples at -20°C has no effect on performance and results of AESKUSLIDES® ANCA Ethanol and ANCA Formalin. #### Procedure: Nine different serum samples have been aliquoted and stored at -20°C for a time period of at least 12 months. Frozen aliquots of the different sera have been thawed and assayed on AESKUSLIDES® ANCA at the indicated time points. All tests have been performed manually according to the IFU and subsequently analyzed at the microscope. | Sample<br>ID | AESKUSLIDES ANCA Ethanol<br>Expected Result | | | AESKUSLIDES ANCA Formalin<br>Expected Result | | | |--------------|---------------------------------------------|---------|----|----------------------------------------------|---------|----| | | Pos/Neg | Pattern | FI | Pos/Neg | Pattern | FI | | 1 | Pos | P | 3 | Pos | C | 3 | | 2 | Pos | C | 3 | Pos | C | 3 | | 3 | Pos | A | 1 | Neg | Neg | 0 | | 4 | Pos | C | 1 | Pos | C | 1 | | 5 | Neg | 0 | 0 | Neg | 0 | 0 | | 6 | Neg | 0 | 0 | Neg | 0 | 0 | | 7 | Neg | 0 | 0 | Neg | 0 | 0 | | 8 | Pos | P | 3 | Pos | P | 2 | | 9 | Pos | P | 3 | Pos | C | 3 | Table 12: Serum sample set for long term serum stability ## Acceptance Criteria: Positive sera have to be found positive and negative sera have to be found negative throughout the whole testing period. Correct patterns have to be found. The fluorescence intensity (FI) is allowed to differ maximum +/- 1 level from the expected value at each test time point. #### Results: Positive samples have been found positive, negative samples have been found negative, and all patterns have been found correctly at all test time points. FI did not differ more than +/- 1 level from the expected values. | AESKUSLIDES ANCA Ethanol | | | | | | | | | | | | | |--------------------------|---------|----|---------|----|---------|----|---------|----|----------|----|----------|----| | Sample<br>ID | month 0 | | month 3 | | month 6 | | month 8 | | month 10 | | month 14 | | | | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | | 1 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | | 2 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | | 3 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | | 4 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | | 5 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 6 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 7 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 8 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | | 9 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Table 13: Long Term Serum Stability AESKUSLIDES ANCA Ethanol {15}------------------------------------------------ | AESKUSLIDES ANCA Formalin | | | | | | | | | | | | | |---------------------------|---------|----|---------|----|---------|----|---------|----|----------|----|----------|----| | Sample<br>ID | month 0 | | month 2 | | month 5 | | month 8 | | month 10 | | month 12 | | | | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | Pos/Neg | FI | | 1 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 2 | Pos | 2 | Pos | 2 | | 2 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | | 3 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 4 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | Pos | 1 | | 5 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 6 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 7 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | Neg | 0 | | 8 | Pos | 1 | na | na | Pos | 2 | na | na | Pos | 1 | Pos | 2 | | 9 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | Pos | 3 | ## Table 14: Long Term Serum Stability AESKUSLIDES® ANCA Formalin ## Conclusion: All acceptance criteria have been fulfilled for the tested sera. The results show that long term storage of sera at -20°C over a time period of at least 12 months has no effect on performance and test results of AESKUSLIDES® ANCA. ## Precision ## a) Within-Lab Precision/Repeatability AESKUSLIDES® ANCA Within-Lab Precision of AESKUSLIDES® ANCA Ethanol and ANCA Formalin processed on HELIOS® (Method A - HELIOS suggestions and Method B - Reader Confirmation of HELIOS images) and processed manually (Method C), based on CLSI Guideline EP12-A2. #### Sample Set 10 samples were assayed on AESKUSLIDES® ANCA Ethanol and Formalin. The sample set includes borderline, low, medium, high positive, and negative samples. The samples are characterized in the following table: | Sample<br>ID | Result | Pattern | Grading | |--------------|--------|----------|---------------------| | 1 | Pos | P | high positive | | 2 | Pos | P | high positive | | 3 | Pos | P | medium positive | | 4 | Pos | P | medium positive | | 5 | Pos | C | borderline positive | | 6 | Pos | C | high positive | | 7 | Pos | C | borderline positive | | 8 | Pos | C | medium positive | | 9 | Neg | negative | negative | | 10 | Neg | negative | negative | #### Table 15: Sample table for Precision study_ANCA Ethanol_Within-Lab Samnle AESKUSLIDES ANCA Ethanol {16}------------------------------------------------ Image /page/16/Picture/0 description: The image is a logo for AESKU.DIAGNOSTICS. The logo consists of a green square with a white image of a zipper inside of it. To the right of the square is the text "AESKU.DIAGNOSTICS" in gray. Below the text is the phrase "THE DIAGNOSTIC TOOL THAT WORKS" in a smaller, lighter gray font. | Sample ID | AESKUSLIDES ANCA Formalin | | | |-----------|---------------------------|----------|-----------------| | | Result | Pattern | Grading | | 1 | Pos | C | medium positive | | 2 | Pos | C | high positive | | 3 | Pos | C | medium positive | | 4 | Pos | C | high positive | | 5 | Pos | C | low positive | | 6 | Pos | C | high positive | | 7 | Pos | C | medium positive | | 8 | Pos | C | high positive | | 9 | Neg | negative | negative | | 10 | Neg | negative | negative | # Table 16: Sample table for Precision study ANCA Formalin Within-Lab ## Procedure 10 samples have been tested on five days, two runs per day, three replicates per sample per run, resulting in 30 data points for each sample. This Within-lab precision study was done for three different Methods: | Method A (HELIOS) | Processing of slides and image recording at HELIOS +<br>HELIOS result suggestion | |-----------------------------------|-------------------------------------------------------------------------------------------------| | Method B (Reader<br>Confirmation) | HELIOS images recorded in Method A + result analysis<br>by two independent readers | | Method C (Manual) | Manual processing of slides and result analysis at the<br>microscope by two independent readers | All runs have been performed according to the respective IFUs. 1 positive and 1 negative control (kit controls) were included in each run. Samples were tested at 1:20 dilution. Results were analyzed by two independent readers. #### Data Analysis Positive/Negative Classification and Pattern was recorded for each sample in each run and for each Method. Pattern suggestion in Method A (HELIOS) is provided by the Vasculitis Pattern Plus software tool. For Method C (Manual performance) fluorescence intensity was also reported. As an estimate of the imprecision of the three methods A, B and C the percentages of positive and negative results will be calculated for each sample: - % Positive (number of positive calls divided by the total number of data points for each sample) - % Negative (number of negative calls divided by the total number of data points per sample) {17}------------------------------------------------ Image /page/17/Picture/0 description: The image contains the logo for AESKU.DIAGNOSTICS. The logo consists of a green square with a white stylized image of a zipper inside. To the right of the square is the text "AESKU.DIAGNOSTICS" in a gray sans-serif font, with the word "AESKU" being larger and bolder than "DIAGNOSTICS". Below the text is the phrase "THE DIAGNOSTIC TOOL THAT WORKS" in a smaller, lighter gray font. Results of the two readers (Method B/C) will be calculated and displayed separately as well as combined. ## Acceptance Criteria Positive sera have to be found positive, and negative sera have to be found negative*. - Reported FI is allowed to differ max. ± 1 level within the study ● - Calculations should meet the following criteria: | Estimate of imprecision | HELIOS | Reader<br>Confirmation | Manual | |-------------------------------------------------------------------|--------|------------------------|--------| | % Positives of positive samples (excluding borderline<br>samples) | > 80% | > 90% | > 90% | | % Negatives of negative samples | > 80% | > 90% | > 90% | borderline samples are samples with analyte concentration near the cutoff. These samples are very low positive. They can be evaluated also negative in certain cases, depending on the reader, the subjective manner of result analysis and normal test variances. For Method A we accept lower percentage of positive/negative results for positive and negative samples, respectively, because we state in the IFU that all results generated by the HELIOS (Method A) must be confirmed by trained personnel. ## Results Results are presented as % positive and % negative results of each sample. Percentages were calculated for the two readers separately (30 replicates for each sample) and for both readers combined (60 data points per sample). All predetermined acceptance criteria were met. | Sample<br>ID | | HELIOS | | User Confirmation | | | | Manual | | | | |--------------|----|--------|-------|-------------------|-------------|-------------|-------------|-------------|-------------|-------------|-------------| | | N | n Neg | n Pos | n Neg | | n Pos | | n Neg | | n Pos | | | | | | | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | | S1 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S2 | 30 | 4 | 26 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S3 | 30 | 1 | 29 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S4 | 30 | 2 | 28 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S5 | 30 | 23 | 7 | 27 | 19 | 3 | 11 | 27 | 19 | 3 | 11 | | S6 | 30 | 3 | 27 | 1 | 1 | 29 | 29 | 0 | 0 | 30 | 30 | | S7 | 30 | 28 | 2 | 19 | 10 | 11 | 20 | 3 | 17 | 27 | 13 | | S8 | 30 | 2 | 28 | 3 | 3 | 27 | 27 | 0 | 0 | 30 | 30 | | S9 | 30 | 26 | 4 | 26 | 28 | 4 | 2 | 30 | 30 | 0 | 0 | | S10 | 30 | 25 | 5 | 30 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | Table 17: Within-Lab Precision AESKUSLIDES ANCA Ethanol number of positive and negative results {18}------------------------------------------------ Image /page/18/Picture/0 description: The image is a logo for AESKU.DIAGNOSTICS. The logo features a green square with a white symbol inside, followed by the text "AESKU.DIAGNOSTICS" in gray. Below the company name, there is a tagline that reads "THE DIAGNOSTIC TOOL THAT WORKS" in a lighter shade of gray. #### Table 18: Within-Lab Precision AESKUSLIDES ANCA Ethanol percentages of positive and negative results | Sample<br>ID | N | HELIOS | | User Confirmation | | | | Manual | | | | |--------------|----|------------|------------|-------------------|-------------|-------------|-------------|-------------|-------------|-------------|----------| | | | % Negative | % Positive | % Negative | | % Positive | | % Negative | | % Positive | | | | | | | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader 2 | | S1 | 30 | 0 | 100 | 0 | 0 | 100 | 100 | 0 | 0 | 100 | 100 | | S2 | 30 | 13.3 | 86.7 | 0 | 0 | 100 | 100 | 0 | 0 | 100 | 100 | | S3 | 30 | 3.3 | 96.7 | 0 | 0 | 100 | 100 | 0 | 0 | 100 | 100 | | S4 | 30 | 6.7 | 93.3 | 0 | 0 | 100 | 100 | 0 | 0 | 100 | 100 | | S5 | 30 | 76.7 | 23.3 | 90.0 | 63.3 | 10.0 | 36.7 | 90.0 | 63.3 | 10.0 | 36.7 | | S6 | 30 | 10.0 | 90.0 | 3.3 | 3.3 | 96.7 | 96.7 | 0 | 0 | 100 | 100 | | S7 | 30 | 93.3 | 6.7 | 63.3 | 33.3 | 36.7 | 66.7 | 10 | 56.7 | 90.0 | 43.3 | | S8 | 30 | 6.7 | 93.3 | 10.0 | 10.0 | 90.0 | 90.0 | 0 | 0 | 100 | 100 | | S9 | 30 | 86.7 | 13.3 | 86.7 | 93.3 | 13.3 | 6.7 | 100 | 100 | 0 | 0 | | S10 | 30 | 83.3 | 16.7 | 100 | 100 | 0 | 0 | 100 | 100 | 0 | 0 | #### Table 19: Within-Lab Precision AESKUSLIDES ANCA Ethanol percentages of positive and negative results for both readers combined | Sample<br>ID | N<br>(Method A) | HELIOS | | N<br>(Method B/C) | User Confirmation | | Manual | | |--------------|-----------------|------------|------------|-------------------|-------------------|------------|------------|------------| | | | % Negative | % Positive | | % Negative | % Positive | % Negative | % Positive | | S1 | 30 | 0 | 100 | 60 | 0 | 100 | 0 | 100 | | S2 | 30 | 13.3 | 86.7 | 60 | 0 | 100 | 0 | 100 | | S3 | 30 | 3.3 | 96.7 | 60 | 0 | 100 | 0 | 100 | | S4 | 30 | 6.7 | 93.3 | 60 | 0 | 100 | 0 | 100 | | S5 | 30 | 77 | 23 | 60 | 76.7 | 23.3 | 76.7 | 23.3 | | S6 | 30 | 10.0 | 90.0 | 60 | 3.3 | 96.7 | 0 | 100 | | S7 | 30 | 93 | 7 | 60 | 48.3 | 51.7 | 33.3 | 66.7 | | S8 | 30 | 7 | 93 | 60 | 10.0 | 90.0 | 0 | 100 | | S9 | 30 | 86.7 | 13.3 | 60 | 90.0 | 10.0 | 100 | 0 | | S10 | 30 | 83.3 | 16.7 | 60 | 100 | 0 | 100 | 0 | #### Table 20: Within-Lab Precision AESKUSLIDES ANCA Formalin number of positive and negative results | | N | HELIOS | | User Confirmation | | | | Manual | | | | |--------------|----|--------|-------|-------------------|-------------|-------------|-------------|-------------|-------------|-------------|-------------| | Sample<br>ID | | n Neg | | n Neg<br>n Pos | | | | n Neg | n Pos | | | | | | | n Pos | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | Reader<br>1 | Reader<br>2 | | S1 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S2 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S3 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S4 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S5 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S6 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S7 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S8 | 30 | 0 | 30 | 0 | 0 | 30 | 30 | 0 | 0 | 30 | 30 | | S9 | 30 | 29 | 1 | 27 | 30 | 3 | 0 | 29 | 26 | 1 | 4 | | S10 | 30 | 27 | 3 | 30 | 27 | 0 | 3 | 30 | 28 | 0 | 2 | {19}------------------------------------------------ Image /page/19/Picture/0 description: The image shows the logo for AESKU.DIAGNOSTICS. The logo consists of a green square with a white graphic inside, followed by the text "AESKU.DIAGNOSTICS" in gray. Below the text is the phrase "THE DIAGNOSTIC TOOL THAT WORKS" also in gray. #### Table 21: Within-Lab Precision AESKUSLIDES ANCA Formalin percentages of positive and negative results | Sample<br>ID | N | HELIOS | | User Confirmation | | | | Manual | | | | |--------------|----|---------------|---------------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------| | | | %<br>Negative | %<br>Positive | % Negative<br>Reader<br>1 | % Negative<br>Reader<br>2 | % Positive<br>Reader<br>1 | % Positive<br>Reader<br>2 | % Negative<br>Reader<br>1 | % Negative<br>Reader<br>2 | % Positive<br>Reader<br>1 | % Positive<br>Reader<br>2 | | S1 | 30 | 0 | 100 | 0 | 0 | 100 | 100 | 0 | 0 | 100 | 100 | | S2 | 30 | 0 | 100 | 0 | 0 | 100 | 100 | 0 | 0 | 100 | 100 | | S3 | 30 | 0 | 100 | 0…