AbIdent Software
Contents
1. 8 P Click the tabs visible at the top of the window to navigate between views 10 1e P 7 Abldent Software 5 2d User Manual 4 1 Sample Info tab The Sample Info tab contains details of the patients examined and additional details of the lab technician performing the test or analysis the kit lots used and the test parameters in the laboratory Note Those fields marked with an asterisk can not be edited using mouse or keyboard Bio Rad Abldent File view Explore Colors About Sample Info Plate 051848 1 R00 Test Parameters Explore Report Bio Rad Medical Diagnostics GmbH IndustriestraBe 1 D 53303 Dreieich Telephone 49 6103 801 807 Abldent Exe Directory C ProgrammesBio Rad BABIDS2D Bio Rad File Directroy C Programme Bio Rad BABID5S2D LotFi les Sample File Directory C sProgrammesBio RadSBABID52D SampleFiles Log File Directory C Programme Bio Rad BABID5S2D Log Export File Directory C Programme Bio Rad BABIDS2D Exports Technician 1 D Sample I D Lot Number Neuland Sample Draw Date Patient Name Test Temp C M Sample Receive Date Patient I D Elisa Test Date Extra Field 1 Date of Report File Extra Field 2 po Patient Phenotype Click to edit ET Fields market with cannot be changed via mouse or keyboard Technician I D If required enter the name of the person in the laboratory2. 27 5 2 2 Setlings Quickstep xcs Bea Emu canons 28 T 1 o P 7 15 Abldent Software 5 2d User Manual 1 Introduction With the aid of this software HLA class and HLA class II antibody identification using the Bio Rad Abldent can be easily and effectively evaluated After data have been evaluated results can be stored and printed out The software has interfaces to all standard microtiter plate readers and automated microtiter plate processors With the aid of the user friendly setup menu laboratory specific adjustments can be carried out quickly and easily 2 Installation 2 1 Minimum system requirements Computer operating system Windows 2000 or better e Screen resolution 1024 x 768 or better e Minimum free harddisk space 20 MB e For importing pipettor data access to pipettor files e g LAN connection e For importing photometer data Serial interface Note If your PC does not meet these minimum requirements this may result in errors extended waiting times and possibly program malfunctions Installation requires Administrator rights Recommendations for connecting up a photometer e Screened serial cable length lt 2m e Transfer rate lt 9600 baud e Do not position cables close to electromagnetic fields e Tuning the parameters in the reader 1e P 7 Abldent Software 5 2d User Manual 2 2 Installation instructions Log on to the
3. 50CBC internal information on the well Line 2 The tick indicates that the well was included in the analysis If more than three 3 wells are excluded from the analysis the software will not be able to evaluate this sample The number 0 918 describes the background adjustment factor BAF for this well The cut off for this well is defined by 2 x the mean of the negative controls multiplied by the BAF This factor compensates for varying reaction strengths in the cells used Line 3 The number 0 457 describes the OD value of the sample after deducting the blank 13 21e P 7 Abldent Software 5 2d User Manual 4 3 Test Parameters tab Test settings are made on the Test Parameters tab TA Bio Rad Abldent File View nfigure Explore Colors About Sample Info Plate 051849 1 R00 Test Parameters Explore Report Limits and Counts Suppressed Items Not included in analysis Positive Cutoff Factor Reset to 2 000 Suppress Patient Phenotype Palen Phet O Negative Cuno Average OD Suppressed Antigens Click to edit 8 243 Mean of 4 negative controlls Cutoff factor x negative mean 0 487000 T Neg Controls Select Using Plate Tab Positive Control Average 0D 1 856 Mean of 1 positive control s Panel Well Select Using Plate Tab Inclusion Cutoff Reset to 50 1 58 1 Antigen or Creg ignored in short tail if positive is less than or equal
4. Anthos HT II DWL Filler set 450 620 Reader data is automatically read in by the software 1e P 7 Abldent Software 5 2d User Manual 4 Working with the software The workflow is as follows e Read in the microplate as a file or using the Reader e use the Reader enter the sample numbers e Evaluate results and edit them e Save results e Print results e Optionally Export results After a pipettor or Reader file has been imported the screen automatically displays the Explore tab showing these results WE Bio Rad Abldent File view nfigure Explore Colors About Sample Info Plate 051849 1 R00 Test Parameters Explore Report Antigen Short Tail Results PRA 16wells 40wells gt 40 0 _ ant Count Sum Inci Cum Inci r Cum r X Cum x Mean 00 1 40 2 14 24 100 100 8 281 0 281 3 158 3 158 6 283 38 3 11 1 23 75 8396 0 271 8 372 2 797 5 523 0 204 35 3 8 1 23 75 80 8 337 8 471 3 978 8 889 0 098 32 1 7 0 24 100 82 8 311 0 526 3 097 11 055 6 122 24 100 0080 0 281 23 75 000 EEI 23 75 000 238 75 000 238 100 000 0 196 100 000 0 196 1 1 IAM maa 196 m Panel List mike OD Sort X suppressed r r g g 8 8 g 8 g B g 8 g g 9 9 9 B 28LB 0 51LB 8 82CBC 62LB 117CBC 98CBC 8 1808CBC 59LB B 21DFW 48LB
5. Class Filenames Abldent Sample File Selection 1 Type in new Sample I D number in the Sample File drop down list Do not type the Class or Class Il Abldent Bio Rad Lot Number part or the file extension ie without the 1 R00 z 12 Click on Bio Rad Lot Number drop down list then select the Bio Rad Lot Number to used with this sample Click OK button J OK x Cancel Abldent Sample File Selection Enter a name for the results file manually Abldent Lot Number Select the Abldent lot you are using 1e P 7 Abldent Software 5 2d User Manual Click OK to generate an empty Reader file You will see a dialogue window which enables you to control the Reader WR Abldent Elisa Module External Program Com1 9600 None 8 2 RTS CTS No XOn X EX Reader Select Reader Options Filter Select Comm Setup About Readers Read Plate J OK Temperature C 23456789 1 R88 Click to change 118986R1 qid No Temp By default an Anthos HT DWL reader is used Refer to chapter 5 1 to find out how to configure the software to use other types of readers Select lt Read Plate gt to initiate the read process In the next window you can enter the current ambient laboratory temperature during the Abldent ELISA tests Select the temperature and click lt OK gt The Bio Rad Reader interface opens Start the read process by clicking lt Start gt Bio Rad Reader Interface 2 1 0 0
6. 27 e 7 Abldent Software 5 2d User Manual 5 2 2 Settings Quickstep The settings for Quickstep are identical to those for Tecan with two minor exceptions e The driver is called Quickstep and the file name is txt Current pipettor driver Quickstep Instrument files mask st 28 Bio Rad Medical Diagnostics GmbH IndustriestraBe 1 63303 Dreieich Tel 449 0 6103 3130 0 Fax 49 0 6103 3130 724 techsupport bmd bio rad com www bio rad com Bio Rad Medical Diagnostics GmbH 2012
7. BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 031023 01IDENT 031023 02IDENT 031023 03 031023 04IDENT 031023 05IDENT 031023 06IDENT 031023 07IDENT 031023 08IDENT 031023 09 031023 10IDENT 031023 11IDENT 031023 12IDENT 031024 01 IDENT 031024 02 031024 03IDENT 031024 04IDENT 031024 05IDENT 031024 06IDENT File nar 031023 031023 031023 031023 031023 031023 031023 031023 031023 031023 031023 031023 031024 031024 031024 031024 031024 031024 w Refresh Setup Abldent f Class C Class ll Lot file 021004 31 P N Start Abldent Exit Select the corresponding files for import by clicking them once with the mouse select the corresponding lot from the Lot file menu and import data by clicking Start import After the import the names of the imported microplates are marked with a green tick and the name is modified This is to prevent the same microplates being imported again iv processed v processed 23 10 2003 23 10 2003 10 24 24 10 32 02 BT_Abld 2x _ver2 BT_Abld 2x _ver2 031023 01IDENT 031023 031023 02IDENT 031023 Note It is important to know the names of the patients which were used when the samples were entered because the software uses th
8. Independent Antigen Statistics means that each specificity is considered and evaluated independently There are therefore no cumulative results This table lists all the specificities on the microplate beginning with the largest number of positive reactions in the sample having a certain specificity There is no relation between the specificities listed as is the case with the Antigen Short Tail Results table and the reactivity of each specificity is assessed individually Antigen Panel List 1e P 7 Abldent Software 5 2d User Manual Antigen Panel List means that each well is assessed individually Wells are listed in the order of their relative OD values The letter X shows whether or not the user has omitted wells from examination Well shows the position of the well and PanID is its internal designation The Relative OD shows how close the OD value is to the cut off value and whether it is above positive rel OD value or below it negative rel OD value A positive rel OD value which is above the cut off value and is still positive after deduction of the cut off value is a positive reaction Antigens shows all the specificities in a well Menu functions in the Antigen Panel List A right click in the Antigen Panel List opens the Panel List Menu window TA Bio Rad Abldent File View Explore Colors About Sample Info Plate 051849 1 R00 Test Parameters Explore Report Antigen Short Tail Results P
9. 2d User Manual 4 5 Report tab Sample result reports can be generated in the Report tab To do so do a right click on the white field The following menu appears Bio Rad Abldent File View gure Explore Colors About Sample Info Plate 051849 1 R00 Test Parameters Explore Report ANTIGEN REPORT 051849 1 R00 05 03 2010 14 16 Bio Rad Medical Diagnostics GmbH Industriestra e 1 D 63303 Dreieich Telephone 49 6103 801 807 Preliminary HLA Class Antigen Report Subject to Supervisor Review Sample ID 651849 1 R60 Techniciai REPORT MENU Patient Name NONE Patient ID NONE Generate Style A Report Ctr A Extra Field 1 NONE Extra Field 2 NON Generate Style B Report Ctrl B Patient Phenntype NONE Beno Style C Report Ctrl C Generate Style D Report Ctrl D DATES FILES Generate Style E Report Draw Date Not Used Lot 021004R1 0ID P Receive Date Not Used Antigen File Comb Ant In Elisa Date 23 10 2003 19 06 13 Creg File Std2 Crg R Ctrl Ww Mean of 4 negative control s 8 243 gt 12 8 257 B12 8 258 C12 Print Report CtrieP Mean of 1 positive control s 1 856 E12 1 856 Suppr Phenotype NO Suppr Antigens NO Test Temp NONE NAW F12 NAW 0 387 MAb NONE bIdent Exe Directory C Programme Bio Rad BABIDS2D Rev 52 4 3 877 Bio Rad File Directroy C Programme Bio Rad BABIDS2D LotFiles Sample File Directory C Programme Bio Rad BABIDS2D Samp
10. 5 ampleFiles instrument dat om Output file format Abldent software C HLA typing AbScreen AbCross software Dutput file header P2 P1 Bio Rad Reader Interface nthos HT II Blanking wells P3 Output to Excel file ucel file nam Simulate with dummy data DUMMY DAT COM setup 25 1e P 7 Abldent Software 5 2d User Manual The required settings for the filter and output file s can be set here The COM settings can be accessed via a different button COM Port Settings Port Be Baud rate 9600 gt Data bits Iss A Parity Evn Flow control Custom Save and leave the Reader changes by clicking OK You currently have a choice of 7 readers Reader setup Reader name Anthos HT Il DWL v Anthos HT Il DWL BioTek ELx800 96 Dynatech MA 5000 Dynex MEX Reference filter Labsystems Ms EX Sample filter Tecan Spectra D wL Dutput file name Note Please also refer to the recommendations relating to Readers in the chapter on System Requirements 26 1e P 7 Abldent Software 5 2d User Manual 5 2 Configuring the pipettor Before you can take over pipettor data the Abldent software has to be configured in such a way that the paths microplate names and microplate assignments tally with the results produced by the pipetter When the software is installed the most typical settings are selected However changes may be
11. UK 189814 05 08 2012 Abldent Software Version 5 2d Copyright c 2012 Bio Rad Medical Diagnostics GmbH All rights reserved 1o P 7 15 Abldent Software 5 2d User Manual LIST OF CONTENTS 1 Introduction Ten innernennnene 2 E Lugeme 2 2 1 Minimum system requirements nennen nnm nnnm rsen nnne neret nennen 2 2 2 Installation me TUC HONS ccce ehe xeu lena enable 3 pe 5 3 1 Iinporting pdatedlots 2 5 aae ene 5 3 2 Importing pipettor fIIGs He ee ee 5 3 3 IMPOMtING areader Tile H X 8 4 Working with the software 10 4 1 Sample Info tab 11 4 2 duri ec 13 4 3 Test Parameters CIUEP c 14 4 4 Explore Ab 16 4 5 Report en 20 4 6 n 21 4 7 SAVING MOS UES m 24 4 8 Exiting the program ese EET 24 EN ee ene ee eee een 25 5 1 Contouring Me ze E nr deren 25 5 2 Configuring the pipettor nn cdd e dei c e e t dd 27 5 2 1 Settings Tecan E
12. at G 12 OD Blanking well found at H 12 OD Blank average 0 005 OD values not calculated 1 2 3 054 085 157 320 418 320 507 070 612 849 918 518 687 958 B81 856 756 889 164 422 179 zanmpmom FPRRRPeNORr orrorror oooroooo 202000902 22000008 22990099 ooooooro ecoooooooo ooor0000 TechID Neuland BleedDate NotUsed ReceiveDate NotUsed TestDate 23 10 2003 19 06 13 SampleID 051849 1 R00 PatientName PatientID ExtraFieldl 23 e 7 Abldent Software 5 2d User Manual 4 7 Saving results If anew sample is imported or the program is closed the user is automatically asked whether the changes are to be saved and in what format Save Sample Data Files CHANGES IN THE DATA MIGHT HAYE INVALIDATED THE REPORT FILES Usually it is best to generate the report files antigen and or creg as the last step prior Save Using New Run Number 1 to saving by overwriting the run number A IF SAVING TO NEW RUN NUMBER Overwrite Run Number 0 Remember that if there are report files they will not exist with the new number unless or until they have been regenerated and saved Cancel This Operation to this new run number B IF OVERWRITING THIS RUN NUMBER Remember to regenerate the report files while the current number is in effect Do Not Save Changes Changes are not saved If the sample is reopened the user will be presented with the default sett
13. template selected check the Settings Is the right software selected check the Settings The Abldent software must be selected Are the correct identification parameters for the test names in Class I Il selected The words entered here and separated from each other by commas must all be contained in the test names in order to display the imported file check Settings Abldent Software 5 2d User Manual Bio Rad Pipettor Interface 2 1 0 0 Bio Rad Quickstep Instrument files folder quickstep dev C Programme Bio Rad BABID52D E sport m Start import Status Date Time Tesnam 2 Plate ID not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed not processed O AAAA PP PE Pe Ei es Pe ERN ERES 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 23 10 2003 24 10 2003 24 10 2003 24 10 2003 24 10 2003 24 10 2003 24 10 2003 Archive imported files 10 24 24 10 32 02 11 00 44 11 08 18 14 04 26 14 12 04 14 40 46 14 48 18 17 30 04 17 37 44 18 06 24 18 13 58 10 21 54 10 29 34 10 58 16 11 05 50 13 52 12 13 59 48 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2 BT_Abld 2x _ver2
14. 8 48LB 1 8 8191 9 9 9 819 89 9 18912 89 98 95 1 8 8 8 5 58 mmm i 0 9 3 35 5 3 1 E PRA The PRA Panel Reactive Antibodies indicates the percentage of all antigen wells on the entire panel that reacted positively The total number of positive wells the example shows 16 is shown as a percentage of the entire panel 40 wells 16 40 x 100 40 0 Antigen Short Tail Results The Antigen Short Tail Results show a statistically calculated HLA antigen identification for the patient The main criterion is the number of correct positive wells for a certain specificity in relation to the total number of wells containing that specificity If this ratio concludes with the specifications in the Test Parameters tab Inclusion Cutoff R Value Cutoff the respective specificity appears in the list The specificity with the most correct positive reactions appears on top of the list 16 7 15 Abldent Software 5 2d User Manual Second place is taken by the specificity with the next best match of correct positive wells in relation to the number of all positive wells for the same specificity Those wells included in the first line are no longer included and are subtracted from the total number of reactions Explanation using patient 21849 as an example 2 of a total of 40 wells contain HLA antig
15. LB 7LB 9 938 21DFW 0 820 SDFW 0 897 51LB 0 866 4 A B B G D D E E Panel File 021004 1 AAA 3 62LB 0 839 27LB 0 919 82CBC 0 818 69LB 0 682 12DFW 0 564 azon This blank line is needed Antigens Cregs tab Antigens and Cregs are listed 5 98CBC 0 879 127CBC 0 813 108CBC 0 828 LR 0 760 117CBC 0 908 6 NC 1 080 NC 1 000 NC 1 808 NC 1 080 PC 1 080 BEALL COMMENTS CAN BE PUT IN THESE COLUMNS 21 Abldent Software 5 2d User Manual FA Bio Rad Abldent File View Configure Explore Colors About Sample Info 021004R1 QID Pan Antigens Cregs Plate 051849 1 R00 Test Parameters Explore Report Antigens DROZ DRO4 EF DROS DROG DRO DROS DROI DR1O DR11 DR12 DR13 DR14 DR1403 DR15 DR16 DR17 DR18 DROX DRWS1 DRWS2 DRWS3 DRWOX pool 82 D003 D004 D005 DQOG ooo pagos D009 553 Cregs 01x 01A 018 10X 19X 28X 02X 024 2B 02C 05X O54 056 osc 07X O74 O7B 07C 08x 084 sB 12X 124 04x 06x b15 b16 b17 b18 b21 b22 b27 b35 b37 b38 b39 05x 08x 02x 05x 12x 07x 07x 05x 12x 08x 08x 1 Creg Panel 05b 08b 02c 05b 12a 07a 07a 05a 04x 08b 08b 5c 05x Bax 06x 07c 05b 04x 06x b3901 88x 08b 06x b3902 88x 08b 06x b40 07x 07b 07c 12x 86x b4005 07x 07b 12x 12a 06x b41 b42 b44 b45 b46 b47 b48 b49 b50 b51 07x 07x 12x 12x 05x 07
16. RA 16wells 40wells gt 40 0 fant icount Sue Hnc cue Inet Cun r X Cue Xo Mean op 40 2 14 24 100 100 5 281 5 281 3 158 3 158 0 283 38 3 11 1 23 75 83 0 271 0 372 2 797 5 523 0 204 35 3 8 1 23 75 50 0 337 0 471 3 978 8 889 0 098 32 1 7 8 24 100 82 0 311 0 526 3 097 11 055 0 122 E 14 0 24 100 8000 a mm 3 4490 8 283 3 13 1 23 75 000 8 238 2 269 8 284 3 13 1 23 75 000 0 238 2 269 0 226 3 13 1 23 75 000 8 238 2 269 8 898 1 15 8 24 100 000 0 196 1 538 8 226 Pr e B53 Ccms CAE MA 63 B44 B35 PANEL LIST SORT BY BSG Tail Antigens Cregs Ctrl T B63 Selected Antigens Cregs Ctri S B64 Relative Optical Density Ctrl O B61 Raw Optical Density Ctrl R Beg Unsorted Ctrl U B45 B45 View Blank Control Wells Ctrl B65 w View Naw amp MAb Wells Ctrl M B51 y Display Raw Optical Density Ctrl Z v Normalize Histogram B45 Increase Cutoff Factor from 2 000 Ctrl I B44 Decrease Cutoff Factor from 2 000 Ctrl D m Reset Cutoff Factor to 2 000 Ctrl F 3 y Use Antigens Ctri A B7E Use Cregs Ctrl C Beg B61 Print Panel Window Ctrl P B44 CO5 CO6 Wo4 W F 06091 B 0 0 0 B g 8 8 g g g g 9 127CBC 0 28LB 0 51LB B S82CBC 62LB B 117CBC S8CBC 8 108C6C Oooocrocoeocoeo ec eo 0979090 90909 9090 0000 e Tail Antigens Cregs highlights the specificities shown in th
17. e Short Tail Results e Under Selected Antigens Cregs individual antigens or Cregs can be highlighted e Relative Optical Density sorts antigen wells by their measured relative optical density e Raw Optical Density sorts antigen wells by their measured values for optical density Unsorted sorts antigen wells the way they are arranged on the microplate Selecting View Blank Control Wells will include the OD values for control wells in the list e Selecting View Naw amp MAb will include the No Antigen Well Naw and the well with only the monoclonal capture antibody MAb only for HLA Class II in the list 18 T 1 o P 7 15 Abldent Software 5 2d User Manual e Display Raw Optical Density shows the measured OD values e Normalize Histogram shows all the OD values in relation to the OD value of the positive control sample If this option is not activated all the OD values will be shown relative to 4 0 Increase or Decrease Cut off Factor from raises or lowers the cut off factor in such a way that the last positive microplate position becomes negative or the first negative microplate position becomes positive e Reset Cut off Factor to 2 000 resets the cut off factor to its default value of 2 000 Selecting either Use Antigens or Use Cregs allows the user to choose between the evaluation method e Print Panel Window prints the Antigen Panel List e 7 Abldent Software 5
18. ens against A31 There was a positive reaction in these two wells The Incl is therefore 100 The columns marked and show the distribution of the reactions This totals 40 The sample is then analysed for further HLA antibodies present in significant numbers One more specificity is listed A28 4 of the remaining 38 wells 40 2 38 on the microplate contain this specificity 3 wells of the sample react as correct positive Ten more positive reactions are evaluated as false positives and one is evaluated as false negative because one A28 reaction did not take place The total number of reactions for specificity A28 is 38 see calculation Since the sample reacts with A28 in 3 out of a total of 4 wells the Incl is 75 The cumulative Incl is higher because it includes the positive reactions of wells covering A31 the total of A31 and A28 specific wells is 6 of which 5 are detected as reactions with correct positives Explanations of the Antigen Short Tail Results table from left to right Antigen name of the specificity or Creg cross reactive group Count the total number of wells covering the respective specificity Sum the total number of wells for identifying one patient Class I 40 wells Class 30 wells correct positive indicates how many wells reacted false positive shows how many wells reacted positive with other antigens 4 false negative shows how many reactions for the
19. ese names and sorts the data in the selection menu for samples to be imported by these names 1e P 7 Abldent Software 5 2d User Manual Click Start Abldent gt to run the Abldent software X Bio Rad Abldent File View Configure Explore Colors About Sample Info Bio Rad Medical Diagnostics GmbH Industriestra e 1 D 63303 Dreieich Telephone 49 6103 801 807 Abldent Exe Directory C Programme Bio Rad BABID5S2D Bio Rad File Directroy C Programme Bio Rad BABIDS2D LotFi less Sample File Directory C Programme Bio Rad BaBIDS2D SampleFi less Log File Directory C sProgrammesBio Rad BABID52D SLog Export File Directory C Programme Bio Rad BABIDS2D Exports Technician I D Sample I D Lot Number Neuland Sample Draw Date Patient Name Test Temp C n E Sample Heceive Date Patient I D mooo Elisa Test Date Extra Field 1 D S585 H ll Date of Report File Extra Field 2 D Patient Phenotype Inactive e re ee Fields market with cannot be changed via mouse or keyboard In the File menu select Start Class Sample or Start Class Il Sample Class Filenames IF STARTING A NEW FILE Abldent Sample File Selection Type in new Sample number in the Sample File drop down list Do not type the Class or Class Il Abldent Bio Rad Lot Number part or the file extension ie without the 1 R00 Click on Bio Rad Lot Number drop down l
20. ick Finish to complete When installation has completed you will see new entries as desktop icons and in the All Programs start menu called Bio Rad Abldent xxxx Reader and Bio Rad Abldent xxxx Pipettor 7 15 Abldent Software 5 2d User Manual 3 Data import 3 1 Importing updated lots Lot specific software updates can be found on the internet at http www medizinische diagnostik dreieich de You can find new lot data by following these steps e Open Download service e Transplantation e Transplantation AbScreen Abldent AbCross A table with these products is shown Choose the required product Abldent Class or Il Click SW of the new lot and then Save A zip folder is downloaded Extract the files in the zip folder to the C Program Files Bio Rad BABID oooQLotFiles folder 3 2 Importing pipettor files To import pipettor data run the Bio Rad Abldent xxxx Pipettor program You will see the Bio Rad Pipettor interface You will see a list of the microplates which can be read in If this list is blank even though you have pipetted some microplates please check the following settings also described in Chapter 5 Is the right path entered under Instrument files folder It is necessary to enter a path here for the exported pipettor files select with the browse button E on the right Is the right pipettor selected check the Settings Is the right instrument file
21. ings as they were when the sample file was imported Save Using New Run Number 2 The number of the run is incremented by one Modifications are saved and the original file is left untouched Overwrite Run Number 0 Modifications are saved under the same file name which means the original file will be overwritten Cancel This Operation Aborts the action without saving modifications 4 8 Exiting the program e Either use the standard Windows method by clicking the in the top right hand corner or e Navigate to File Exit 24 1e P 7 Abldent Software 5 2d User Manual 5 Appendix 5 1 Configuring the reader You may configure the Reader settings This is normally the task of the Bio Rad product specialists If you modify these settings without contacting Bio Rad first Bio Rad cannot accept any guarantee for error free functioning of the interface with the Reader Navigate to File Start Class Sample or Start Class Il Sample manual entry of the microplate number and selection of the lot number select lt Read Plate gt you will see the Reader window Bio Rad Reader Interface 2 1 0 0 E3 Anthos HT II DWL Setup Filter set 450 520 Exit Select Setup to access the setup menu Reader setup Reader name Anthos HT Il DWL v Set 1 Set 2 Set 3 Sample filter 450 370 wavelength Reference filter 520 492 wavelength Output file name c SProgramme Bio R ad BABIDS52D
22. ist then select the Bio Rad Lot Number to used with this sample 3 Click OK button IF OPENING AN EXISTING FILE J x Cancel 1 Select filename from Sample File drop down list 2 Click OK button Abldent Sample File Selection Select the sample names from a list Abldent Lot Number Select the Abldent lot you are using Click OK to confirm e 7 Abldent Software 5 2d User Manual 3 3 Importing a reader file To import reader data run the Bio Rad Abldent xxxx Reader program The Bio Rad Adldent software will start WE Bio Rad Abldent File View Configure Explore Colors About Sample Info Bio Rad Medical Diagnostics GmbH Industriestra e 1 D 63303 Dreieich Telephone 49 6103 801 807 Abldent Exe Directory C Programme Bio Rad BABID52D Bio Rad File Directroy C Programme Bio Rad BABIDS2D LotFi less Sample File Directory C Programme Bio Rad BABIDS2D SampleFi les Log File Directory C sProgrammesBio Rad BABID52DSLog Export File Directory C Programme Bio Rad BABIDS2D Export Technician I D Sample I D Lot Number Neuland Sample Draw Date Patient Name Test Temp I ee Patient I D p Elisa Test Date Extra Field 1 EE Date of Report File Extra Field 2 ee Patient Phenotype Inactive Fields market with cannot be changed via mouse or keyboard In the File menu select Start Class Sample or Start Class Il Sample
23. leFi less Antigen Short Tail Results PRA l6wells 48wells gt 40 0 Antigen Cnt Sum CIn Cum r 431 48 2 14 100 100 28 281 r 428 3 1 75 83 ar 312 cas 3 8 75 80 Be IE DA 471 r B41 m 100 82 811 526 r B18 2 67 79 345 578 r B56 1 100 80 414 2632 T 3 100 81 471 r 688 r ns and or r values are their cutoff values B57 Remaining inclusions V Oy CU LU NJ CU There are five possible displays to choose from Report Style A and Report Style B These report types contain first of all the Antigen Panel List and below that the Antigen Short Tail Results The difference between Type A and Type B is the target printer laser or colour printer Report Style C This report type contains Antigen Short Tail Results first followed by the Antigen Panel List Report Style D This report type only includes the Antigen Short Tail Results Printout Results are printed by right clicking and selecting Print Report 20 Abldent Software 5 2d User Manual 4 6 Other tabs In the View menu further tabs can be displayed besides the tabs shown by default chapter 4 1 to 4 5 Lot Number Qid Pan tab The lot specific microplate assembly is displayed IA Bio Rad Abldent File View Faure Explore Colors About Sample Info 021004R1 QID Pan Plate 051849 1 R00 Test Parameters Explore Report BID File 021004R1 01D 1 2 58CBC LA 0 918 0 877 48
24. lors About Sample Info Plate 051843 1 R00 Test Parameters Explore Report Comment Portion of Elisa File Begin each line with at least one space 021004R1 0ID 8051849 1 R00 Bio Rad Pipettor Interface version 2 1 0 0 Test limit 3 000 Test wavelengths 405nm 492nm Created from file 8031823 11IDENT TXT Assay name BT AbId 2x I ver2 Blanking well found at G 12 0 Blanking well found at H 12 0 Blank average 0 005 OD values not calculated 4 5 D 8 8 D 0 0 i 50CBC L 59LB 98CBC V 8 918 V 0 877 5 V 0 740 V 0 879 0 242 0 457 0 250 0 256 0 348 48LB 7LB 1BW 127CBC V 1 828 V 8 938 jw 8 811 v 0 813 8 382 6 319 8 278 1 409 8 365 BF 21DFW 2HC 188CBC V 0 891 v 8 828 T V 8 987 V 0 828 0 590 9 284 B 355 0 339 9 312 15LB LR jv 0 816 v 8 897 jv 8 792 V 8 768 8 258 8 517 8 248 8 883 95CBC 51LB 117CBC jw 8 926 VW 8 866 jw 8 988 0 629 2BW E jw 8 847 V 0 918 v 1 008 0 469 8 678 8 305 8 218 0 100 17LB 11CBC 36CBC lv 8 789 v 1 881 0 890 5 VW 1 088 8 234 0 270 8 537 9 843 9 239 8 004 19DFW 53LB 103CBC jw 0 691 VW 0 752 V 0 852 w 8 863 VW 1 006 0 580 90 288 0 269 0 378 0 005 The information on the individual wells on the microplate is shown below by reference to one well well A7 Line 1
25. made as required This is normally the task of the Bio Rad product specialists If you modify these settings without contacting Bio Rad first Bio Rad cannot accept any guarantee for error free functioning of the interface with the pipetter Our customer services staff will be pleased to assist you 5 2 1 Settings Tecan After starting the program version Bio Rad Abldent xxxx pipetter selecting the Setup option takes the user to the dialogue screen for pipetter settings The required settings can now be made Pipettor setup Current pipettor driver Pipettor name Tecan v Tecan RMP ok Instrument files mask Output file name imp dat C Programme Bio Rad BABID52D 4S ampleFiles instrument dat Cancel C HLA Typing Software Auto exit program after import AbScreen AbCross Score Interpretation Abldent Software Test name Class JST ID I Test name Class Il aT ID I Ic Programme Bio Rad BABID52D Abl dent uro exe m Abldent lot file path C Programme Bio Rad BABID52D LotFiles ad Blank correcting factor gt C Subtract blank from OD values f No blank subtraction Pipettor name Enter the name of the pipetter Data input mask Tecan rmp dat Quickstep txt Beplll Input file path normally the import folder of the Abldent software Output file name This file is generated by the software itself It is expected to be in the import folder and must be named instrument dat
26. performing the analysis By default the computer enters the name of the current Windows user This can be changed Sample Draw Date The date when the blood sample was taken can be entered here Sample Receive Date The date when the sample reached the laboratory can be entered here 1o P 7 15 Abldent Software 5 2d User Manual ELISA Test Date The date the Abldent test was performed appears here Date of Report File The date the report is generated appears here once it has been generated Sample I D The sample number appears here Patient Name Patient I D The name or lab ID of the patient may be entered here Extra Fields 1 and 2 Freeform text comments can be entered here Abldent Lot Number The Lot Number of the Abldent lot used is displayed here Test Temp C The ambient temperature in the laboratory during the test can be entered here Patient Phenotype Click lt Patient Phenotype gt to enter the patient s HLA type To do so you may mark the corresponding specificities in the list Click lt OK gt to confirm If the patient s phenotype is entered here these specificities may be suppressed on the Test Parameters tab 1e P 7 Abldent Software 5 2d User Manual 4 2 Plate tab The Plate tab contains the microplate layout of the Abldent lot used The file name is shown in brackets e g 7135 1 R00 The R stands for Raw Data Bio Rad Abldent DAR File View Configure Explore Co
27. respective specificity did not take place o correct negative shows how many reactions were negative for other specificities Incl if the number Cnt matches with the number this makes 100 Incl is calculated using the following equation Incl Cnt x 100 Cum Incl 2 cumulated Incl this value is equal to the Incl value for the first position of the list but higher in line two because it includes the reaction of the specificity in line one and so on shows the reproducibility of test results An R value of e g 0 6 indicates that 60 of the results are correct positive and 4096 non specific Explanation using an example If 13 false positive reactions were included with 2 correct ones the value of R is 0 296 If there are no non specific false positive or false negative reactions the value of R will be exactly 1 000 Cumr cumulative value for R is the same as the specificity at the top of the table with the second specificity the reactions of the first specificity are included and so on nye means chi squared which is a statistical measurement of the credibility of the test results The higher the value of chi squared the more accurate and credible the meaning of the test results Cum X cumulative value of chi squared Avg OD average OD The difference between Rel OD and Avg OD can be seen from the following formula Rel OD Avg OD well cut off Independent Antigen Statistics
28. sis Note For this purpose the patient s HLA type data must be entered on the Sample Info tab Suppressed Antigens Certain HLA specificities can be excluded from the analysis here Neg Controls Panel Well This option displays the microplate positions deactivated in the Plate tab In the Antigen and Creg Files window you can select whether you wish to analyse using individual HLA antigens or Cregs Cross Reactive Groups 1e P 7 Abldent Software 5 2d User Manual 4 4 Explore tab The Explore tab is subdivided into 3 sections describing the results of the Abldent ELISA WE Bio Rad Abldent File View r Explore Colors About Sample Info Plate 051849 1 R00 Test Parameters Explore Report Antigen Short Tail Results PRA 16wells 40wells gt 40 0 Jant Count Sum Inci Cum Inct r Cum r x Cum x Mean 00 1 40 2 14 0 24 100 100 0 281 0 281 3 158 3 158 0 283 38 3 11 1 23 75 83 0 271 0 372 2 797 5 523 0 204 35131611 193 75 50 0 337 0 471 3 978 8 889 0 098 32 1 7 8 24 100 82 0 311 0 526 3 097 11 055 9 122 24 100 800 0 281 23 75 000 238 23 75 000 238 23 75 000 238 24 100 800 0 196 24 100 000 0 196 1 m 24 IMM mmm m 196 e es s g g g g g B g g g g g B g 127CBC_ 0 28LB 8 51LB 8 82CBC 62LB 117CBC 8 98CBC 8 188CBC 8 8 21DFW
29. system as the Administrator Insert the CD in the CD drive Run the BABIDsetup exe program from the CD 1 First you will see the Installation Wizard followed by the welcome message i Bio Rad Abldent 5 2d InstallShield Wizard Welcome to the InstallShield Wizard for Bio Rad AbIdent 5 2d The InstallShield R Wizard will install Bio Rad AbIdent 5 2d on your computer To continue click Next WARNING This program is protected by copyright law and international treaties Click lt Next gt to display the following sequence of windows 2 Licence agreement Select accept the terms in the license agreement The Next button cannot be clicked until you do so 3 Note on installed files Next 4 PC user and company lt Next gt 5 Suggested target folder The default folder is C Program Files Bio Rad BABID ooo xxxx current version Changing the target folder Enter a changed folder name in this window or simply click Next to confirm the default 6 Overview of settings before the installation itself begins Click Install to confirm the installation will then begin 1e P 7 Abldent Software 5 2d User Manual 8 When installation is complete you will see the following message 5 Bio Rad Abldent 5 2d InstallShield Wizard InstallShield Wizard Completed The InstallShield Wizard has successfully installed Bio Rad AbIdent 5 2d Click Finish to exit the wizard Finish Cancel Cl
30. to this number ari Cre Fior R Yalue Cutoff _ Reset to 0 25 Antigen File p 25 Antigen or Creg ignored in short tail if correlation is less than or equal to this number Comb Ant Use Antigens Creg Definition File Notes std2 Crg C Use Cregs Changes on this page valid for this sample only Use configure pull down menu for persistent changes Test specific parameters can be set under Limits and Counts Positive Cut off Factor The value by which the mean value of the negative controls is to be multiplied in order to calculate the cut off Negative Control Average OD Calculation of the mean values from four negative controls after deducting the blank Positive Control Average OD Calculation of the positive control sample after deducting the blank Inclusion of cut off The inclusion cut off is recalculated for each sample and indicates that an antigen or Creg Cross reactive group is no longer included in the short tail list if this would mean that the overall percentage of 50 196 is not attained Tz 1 o P 7 15 Abldent Software 5 2d User Manual R value cut off The R value cut off indicates that if the R value is e g 0 25 or less that antigen or Creg is no longer included in the short tail list In the Suppressed Items window certain specificities can be excluded from the analysis Suppress Patient Phenotype Here the patient s HLA specificities can be excluded from the analy
31. x 07x 05x 05x 05x 07b 07a 12a 12a 05b 07a 07b O5c 12x 05a 07c 12x 86x 07b 86x 04x 06x o5c 07c 12x 84x 07c 12x 86x 12x 12a 84x 12a 86x 05b 84x b5102 85x 05a 05b 84x b5103 85x 05a 05b 84x b52 b53 b54 b55 b56 b57 05x 05x 07x 07x 07x 02x 05a 05a 07a 07a 07a 02c 05b 12a 04x 05b 04x 06x 06x 06x 05x 5c 04x 22 21e P 7 Abldent Software 5 2d User Manual e Creg Definition tab Antigens are assigned to the respective cregs Bio Rad Abldent File View Configure Explore Colors About Sample Info Creg Def Std2 Crg Plate 051849 1 R00 Test Parameters Explore Report Creg Definition File 3 Max length of cregs This blank line is needed STANDARD CREGS and Component Antigens s Set 10 06 00 01x 01a 515 10x 19x 28x 82x 02c 05x 05a 05b O5c 07x 07a 07b 07c 08x 08a 08b 12x 12a 04x 06x Blx 81a 81b 02x 2c 02x 2c 02x 2c Blx 81a 01b 02x 01x 0lb Blx 813 815 01x e ELISA Data File tab Reader data for the tested sample is shown P Bio Rad Abldent File View Configure Explore Colors About Sacre 051849 1 R00 Plate 051849 1 R00 Test Parameters Explore Report Elisa Reader Data 021004R1 0ID 051849 1 R00 Bio Rad Pipettor Interface version 2 1 0 0 Test limit 3 000 Test wavelengths 485nm 492nm Created from file 031823 11IDENT TXT Assay name BT bId 2x I ver2 Blanking well found
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