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Instructions for Use Altostar Parvovirus altona DIAGNOSTICS Instructions for Use AltoStar® Parvovirus B19 PCR Kit 1.5 09/2020 EN ® AltoStar AltoStar® Parvovirus B19 PCR Kit 1.5 For use with CFX96™ Deep Well Dx System (Bio-Rad) AS0101513 96 09 2020 MAN-AS0101510-EN-S02 altona Diagnostics GmbH • Mörkenstr. 12 • D-22767 Hamburg AltoStar® Parvovirus B19 PCR Kit 1.5 Content 1. About these Instructions for Use .............................................................8 2. Intended Use ............................................................................................... 9 3. Kit Content .................................................................................................. 9 4. Storage and Handling .............................................................................. 10 4.1 Storage .................................................................................................10 4.2 Handling ............................................................................................... 11 4.2.1 Master A and Master B .........................................................................12 4.2.2 QS and NTC .........................................................................................12 5. Background Information .........................................................................12 6. Product Description ................................................................................. 13 6.1 Master A and Master B .........................................................................14 6.2 Quantification Standards ......................................................................14 6.3 No Template Control .............................................................................14 6.4 AltoStar® Workflow ...............................................................................15 7. Samples ....................................................................................................15 7.1 Sample Types .......................................................................................15 7.2 Sample Collection and Handling ..........................................................16 7.3 Sample Volume ....................................................................................16 7.4 Sample Tubes.......................................................................................17 7.5 Sample Barcodes .................................................................................17 8. Material and Devices Required, but not Provided ................................ 17 9. Warnings, Precautions and Limitations ................................................. 20 10. Procedure ................................................................................................. 22 10.1 Overview of the AltoStar® Workflow ......................................................22 4 AltoStar® Parvovirus B19 PCR Kit 1.5 10.2 Starting the AltoStar® AM16 ..................................................................26 10.3 Performing Maintenance ......................................................................27 10.4 Programming an AltoStar® Run ............................................................29 10.4.1 Manual Programming ...........................................................................29 10.4.2 Importing from LIMS .............................................................................36 10.5 Creating an AltoStar® Run ....................................................................37 10.6 Starting a Purification Run ....................................................................39 10.6.1 Sample Preparation ..............................................................................40 10.6.1.1 Plasma .......................................................................................40 10.6.2 Preparing Reagents for a Purification Run ........................................... 41 10.6.3 Instrument Loading for a Purification Run ............................................ 42 10.7 During the Purification Run...................................................................53 10.8 End of the Purification Run ...................................................................55 10.9 Purification Run Results .......................................................................58 10.10 Eluate Stability ......................................................................................60 10.10.1 Storage .................................................................................................60 10.10.2 Sealing of the Eluate Plate ...................................................................60 10.10.3 Unsealing of the Eluate Plate ...............................................................62 10.11 Starting a PCR Setup Run....................................................................62 10.12 Preparing Reagents for a PCR Setup Run ........................................... 64 10.12.1 Loading the AltoStar® AM16 for a PCR Setup Run ............................... 65 10.13 During the PCR Setup Run ..................................................................73 10.14 End of the PCR Setup Run...................................................................74 10.15 PCR Setup Run Results .......................................................................76 10.16 Sealing of the PCR Plate ......................................................................78 10.17 PCR Mix Stability ..................................................................................79 10.18 Starting a PCR Run ..............................................................................80 10.19 During the PCR Run .............................................................................83 10.20 Assigning Assays to Well Groups .........................................................84 5 AltoStar® Parvovirus B19 PCR Kit 1.5 10.21 PCR Data Analysis ...............................................................................87 10.21.1 Baseline Correction ..............................................................................88 10.21.2 Exclusion of Irregular PCR Signals ......................................................90 10.21.3 Setting of Thresholds ............................................................................94 10.21.4 Exclusion of Wells Containing Invalid Data .......................................... 97 10.21.4.1 Validity of a Diagnostic PCR Run (qualitative) ......................... 101 10.21.4.2 Validity of a Diagnostic PCR Run (quantitative) ....................... 101 10.21.4.3 Validity of Results for a Sample ................................................ 102 10.21.5 Export of PCR Results for Automated Result Interpretation ............... 103 10.21.6 Export of PCR Results for Manual Result Interpretation .................... 105 10.21.6.1 Manual Interpretation of Results .............................................. 107 11. Disposal .................................................................................................. 110 12. Performance Evaluation ........................................................................ 111 12.1 Plasma.................................................................................................111 12.1.1 Analytical Sensitivity ............................................................................111 12.1.2 Analytical Specificity ........................................................................... 112 12.1.2.1 Negative Samples .................................................................... 113 12.1.2.2 Interfering Substances ............................................................. 113 12.1.2.3 Cross Reactivity ....................................................................... 113 12.1.3 Linear Range ...................................................................................... 114 12.1.4 Precision ............................................................................................. 115 12.1.5 Total Failure Rate ............................................................................... 116 12.1.6 Carry Over .......................................................................................... 116 12.1.7 Diagnostic Evaluation ......................................................................... 117 13. Quality Control ....................................................................................... 119 14. Technical Assistance ............................................................................. 119 15. Literature ................................................................................................. 119 6 AltoStar® Parvovirus B19 PCR Kit 1.5 16. Trademarks and Disclaimers ................................................................120 17. Explanation of Symbols ........................................................................121 7 AltoStar® Parvovirus B19 PCR Kit 1.5 1. About these Instructions for Use These Instructions for Use guide the user in utilizing the AltoStar® Parvovirus B19 PCR Kit 1.5 on the AltoStar® Automation System AM16 (Hamilton; in the following summarized as AltoStar® AM16) with the AltoStar® Connect software (Version 1.6.16 or higher, Hamilton) for automated PCR setup and on the CFX96™ Deep Well Dx System* (Bio-Rad, in the following summarized as CFX96™ DW Dx) with the CFX Manager™ Dx software (Version 3.1, Bio-Rad) for real-time PCR. The main operation steps of the complete "AltoStar® Workflow" (for details see chapter 6.4 AltoStar® Workflow) are described
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