Change to insert re sensitivity added EFG (Sect 1)
Contents
1. MININEPH IgG LOW LEVEL KIT For In vitro Research Use Product Code ZK004 LL Product manufactured by The Binding Site Group Ltd 8 Calthorpe Road Edgbaston Birmingham B15 1QT UK www bindingsite co uk Telephone 44 0 121 456 9500 Fax 44 0 121 456 9749 e mail info bindingsite co uk MININEPH and MININEPHPLUS are trademarks of The Binding Site Group Ltd Birmingham UK 1 INTENDED USE This kit is designed for the n Vitro measurement of IgG at low levels using the MININEPH or MININEPHPLUS When using neat sample the approximate measuring range is 10 5 168mg L The assay is for Research use only it is the responsibility of the user to validate the assay and ensure that it is fit for purpose The MININEPHPLUS analyser is not available in the USA 2 PRINCIPLE OF THE ASSAY The determination of soluble antigen concentration by nephelometric methods involves a reaction with the antibody to form insoluble complexes When light is passed through the suspension formed a portion of the light is scattered and detected by a photodiode The amount of light scattered is directly proportional to the specific protein concentration in the test sample Concentrations are automatically calculated by reference to a calibration curve stored within the instrument 3 REAGENTS 3 1 MININEPH HUMAN IgG LOW LEVEL REAGENT This has been adsorbed to monospecificity for IgG and is supplied in stabilised liquid form2. opened the reagent and the control should be stored at 2 8 C and the buffer at room temperature Opened reagent buffer and control are stable for 12 weeks when stored as recommended The On Board Buffer 1 should be stored at room lemperature Opened On Board Buffer 1 is stable for 4 weeks when stored as recommended 6 SPECIMEN COLLECTION AND PREPARATION Specimen collection preparation and storage have not been validated for this kit It is the responsibility of the user to validate this aspect of the procedure v4 METHODOLOGY 7 1 MATERIALS PROVIDED 1 x 2 mL MININEPH Human IgG Low Level Reagent 1 x 25 mL MININEPH IgG Low Level Buffer 1 x 0 5 mL MININEPH Human IgG Low Level Control Magnetic swipe card containing lot specific calibration information Quality Control Certificate Instruction leaflet NNNNNN SE cathy cael aac gs gt oonRworyo 7 2 MATERIALS REQUIRED BUT NOT PROVIDED MININEPH Te MININEPH instrument AD200 7 2 2 MININEPH printer AD210 optional 7 2 3 MININEPH reagent accessory pack ZK500 R 7 2 4 Electronic pipette e g AD205 7 2 5 Pipette 5 40uL 7 2 6 Equipment for the collection and preparation of test samples 7 3 MATERIALS REQUIRED BUT NOT PROVIDED MININEPHPLus 7 3 1 MININEPHPLUus instrument AD500 C D E 7 3 2 MININEPHPLUS PRINTER AP1310DPK1T63 optional 7 3 3 Bar Code Reader optional 7 3 4 MININEPH reagent accessory pack ZK500 R 7 3 5 Pipette 5 1000uL 7 3 6 Equipment for t
3. It contains 0 099 sodium azide 0 1 EACA and 0 01 benzamidine as preservatives 3 2 MININEPH IgG LOW LEVEL SWIPE CARD This is encoded with details of the reaction curve specific to the respective lot of reagent This card is lot specific and must be used only with this lot of reagent The curve on this card has been prepared using secondary calibration materials that have been calibrated against CRM470 3 3 MININEPH IgG LOW LEVEL BUFFER For use with this lot of IgG Low Level reagent only Contains 0 099 sodium azide as a preservative 3 4 MININEPH HUMAN IgG LOW LEVEL CONTROL This consists of pooled normal human serum and is supplied in stabilised liquid form It contains 0 099 sodium azide 0 1 EACA and 0 01 benzamidine as preservatives The acceptable range of IgG concentrations are stated on the Quality Control Certificate included in the kit The lot number quoted on the Quality Control Certificate should be identical to the kit lot number 4 All donors of human serum supplied in this kit have been serum tested and found negative for hepatitis B surface antigen HBsAg and antibodies to human immunodeficiency virus HIV1 and HIV2 and hepatitis C virus The assays used were either approved by the FDA USA or cleared for in vitro diagnostic use in the EU Directive 98 79 EC Annex ll however these tests cannot guarantee the absence of infective agents Proper handling and disposal methods should be established as for all potentially i
4. V Day to day Inter instrument mg L n 30 CV n 30 CV n 15 Serum 1 151 8 3 85 7 18 3 45 Serum 2 38 8 2 84 3 01 6 84 These data represent the average coefficient of variation CV of three within batch measurements repeated ten times at each concentration Ten within batch measurements were performed on three separate occasions and the overall CV for the thirty results at each concentration calculated Assays were performed five times at each concentration on three instruments The overall CV of the fifteen results at each concentration was calculated 10 2 COMPARISON STUDIES 10 2 1 MININEPHPLUS 20 normal adult sera and 17 clinical adult sera were tested on the MININEPH and MININEPHPLUS The study demonstrated a good agreement yielding the following Passing amp Bablok equation and linear regression correlation coeffiecient y 1 05x 3 51 y MININEPHPLUus IgG Low Level x MININEPH IgG Low Level correlation coefficient r 0 998 Insert Code ZINO04 L Version 19 September 2011 Page 2 of 2
5. be used 7 5 15 Place cuvette in chamber Place a cuvette containing a stirring bar and 40uL of diluted sample in the cuvette chamber Press the cuvette down gently until it reaches the bottom of the chamber The cuvette will be detected automatically 7 5 P 7 5 16 Supplementary buffer Using the MININEPHPLUs hand held pipette aspirate 400uL of MININEPH IgG Low Level buffer 7 5 17 Air Gap Using the MININEPHPLUs hand held pipette aspirate an air gap 7 5 18 Aspirate Reagent Using the MININEPHPLUS hand held pipette aspirate 40uL of MININEPH Human IgG Low Level antiserum 7 5 19 Add Reagent Dispense the aspirated reagents into the cuvette The stirring bar will rotate and the assay will begin After a 30 second blanking time the assay will take 152 seconds to complete The result will be displayed Results will be automatically printed if a printer is connected 7 5 20 If the instrument indicates the result is higher than the standard measuring range reassay the sample at a higher dilution of 1 11 400uL of MININEPH Sample Diluent 40L of sample using the electronic pipette The sample dilution should be enetered as 1 11 see section 7 4 9 If the results is still displaying as higher than the measuring range the sample should be reassayed at a higher dilution of 1 121 400uL of MININEPH Sample Diluent 40uL of sample diluted 1 11 The sample dilution should be enetered as 1 121 7 5 21 When all assays for the chosen chemistry num
6. ber have been completed press esc and select the chemistry number for the next set of assays 7 5 22 Empty waste pot and discard the pipette tip from the hand held pipette 7 6 As with all good laboratory practice users should run the control with every batch of samples The control included in this kit should be treated in the same way as samples 8 INTERPRETATION OF RESULTS 8 1 Results are calculated by the instrument and displayed in mg L If a printer is attached the result is automatically printed out together with the patient identification code and the sample dilution Further calculations are not necessary 8 2 The assay range is limited to the ranges stated below Dilution Range Neat 10 5 168 mg L 1 11 115 5 1848 mg L 1 121 1270 5 20328 mg L The assay has an antigen excess capacity of four times the top of the range at any given sample dilution for example sample concentrations up to at least 672mg L will not result in antigen excess when using neat sample Higher concentrations may give misleading results if this is suspected samples should be reassayed using the next sample dilution in the above series 9 LIMITATIONS OF PROCEDURE 9 1 SPECIFIC TEST LIMITATIONS 9 1 1 Nephlometric assays are not suitable for measurement of highly lipaemic or haemolysed samples or samples containing high levels of circulating immune complexes CICs due to the unpredictable degree of non specific sca
7. he collection and preparation of test samples 7 3 7 MININEPHPLUs On Board Buffer 1 SN107 7 3 8 Pipette tips for use with the MININEPHPLUs refer to MININEPHPLUS User Guide 7 4 TEST PROCEDURE MININEPH ANALYSER 7 4 1 Summary of reagent volumes added to the cuvette Reagent Volume added Sample Undiluted 40uL MININEPH IgG Low Level Buffer 400uL MININEPH Hu IgG Low Level Reagent 40uL 7 4 2 Switch the analyser and printer if attached on 7 4 3 Enter chemistry number Enter the chemistry number IgG Low Level 3 and press enter 7 4 4 Swipe chemistry card This message will only be displayed if this chemistry has never been used before or you wish to change antiserum lot number Pass the swipecard through the swipecard reader moving from the front of the instrument to the back The magnetic strip should be at the bottom facing left 7 4 5 Check reagent lot number Press enter 7 4 6 IgGLL lot xxxx OK 1 Y 2 N Compare the details displayed with those on the antiserum label If the lot number displayed is identical to that printed on the antiserum vial select YES press 1 and continue to step 7 4 7 If the vial lot number is different from that displayed select NO press 2 and return to step 7 4 4 to allow the details of the correct batch to be entered 7 4 7 Prepare one MININEPH cuvette for each sample to be assayed Using the forceps provided with the MININEPH place a stirring bar in each cuvette and then us
8. ing a pipette add 40uL of sample carefully to the bottom of each cuvette 74 8 Enter sample ID Enter an identity code e g 1 for the first sample to be assayed then press enter to continue refer to user manual for choice of identity codes 7 4 9 Sample dilution 1 1 undiluted Accept the recommended dilution by pressing enter or type in a new dilution factor if an alternative dilution is to be used 7 4 10 Place cuvette in chamber Place a cuvette containing a stirring bar and 40uL of sample in the cuvette chamber Press the cuvette down gently until it reaches the bottom of the chamber The cuvette will be detected automatically 7 4 11 Add reagent Fill an electronic pipette with 400uL of MININEPH IgG Low Level buffer and 40uL of MININEPH Hu IgG Low Level antiserum and dispense its contents into the cuvette The MININEPH will detect the addition followed by movement of the stirring bar and the assay will begin It is not necessary to press enter After a 30 second blanking time the assay will take 155 seconds to complete the result will then be displayed and printed automatically if a printer is connected 7 4 12 On completion of the assay remove the cuvette and press enter to perform the next assay 7 4 13 If the instrument indicates the result is higher than the intended measuring range reassay the sample at a higher dilution of 1 11 400uL of MININEPH Sample Diluent 40L of sample using the electronic pipette The sample dilution sho
9. nfective material including but not limited to users wearing suitable protective equipment and clothing at all times Only personnel fully trained in such methods should be permitted to perform these procedures WARNING This product contains sodium azide and must be handled with caution suitable gloves and other protective clothing should be worn at all times when handling this product Do not ingest or allow contact with the skin particularly broken skin or open wounds or mucous membranes If contact does occur wash with a large volume of water and seek urgent medical advice Explosive metal azides may be formed on prolonged contact of sodium azide with lead and copper plumbing on disposal of reagent flush with a large volume of water to prevent azide build up This product should only be used by suitably trained persons for the purposes statedin the Intended Use Strict adherence to these instructions is essential at all times Results are likely o be invalid if parameters other than those stated in these instructions are used Reagents from different batch numbers of kits are NOT interchangeable If large numbers of ests are performed care should be taken to ensure that all reagents are from the same batch 5 STORAGE AND STABILITY The unopened kits should be stored at 2 8 C and can be used until the expiry date given on he kit box label DO NOT FREEZE The buffer should be allowed to equilibrate to room emperature prior to use Once
10. tter these sample types may generate Unexpected results should be confirmed using an alternative assay method 9 1 2 Results may be adversely affected by fragmentation or aggregation of IgG It is the responsibility of the user to assess the integrity of the IgG and validate its effect upon results 9 1 3 Assay linearity may be affected by the clonality of the IgG It is the responsibility of the user to validate the assay for each clone 9 1 4 Results may be affected by the constitution of the sample media It is the responsibility of the user to validate the assay for the type of sample media being used 9 2 TROUBLE SHOOTING Problem Possible Causes s Suggested Action s Error message Blank too high reassay displayed Controls out of range Test sample giving unexpectedly low result Very high analyte concentration Turbid samples Product deterioration Operator error Antigen excess Reassay sample at a higher dilution Clarify before use Check expiry date Repeat assay with the correct sample dilution Repeat assay at higher dilution Check if the two results agree 10 PERFORMANCE CHARACTERISTICS 10 1 Precision 10 1 1 Precision MININEPH IgG Precision Summary Mean mg L Intra batch CV n 30 Serum 1 146 6 2 99 Serum 2 44 4 2 47 10 1 2 Precision MININEPHPLUS IgG Precision Summary Mean Intra batch C
11. uld be enetered as 1 11 see section 7 4 9 If the results is still displaying as higher than the measuring range the sample should be reassayed at a higher dilution of 1 121 400uL of MININEPH Sample Diluent 40L of sample diluted 1 11 The sample dilution should be enetered as 1 121 7 4 14 When all assays for the chosen chemistry have been completed press escape esc and select the chemistry number for the next set of assays 7 5 TEST PROCEDURE FOR MININEPHPLUS ANALYSER Wda Summary of reagent volumes added to the cuvette Reagent Volume added Sample Undiluted 40uL MININEPH IgG Low Level Buffer 400uL MININEPH Hu IgG Low Level Reagent 40uL 7 5 2 Ensure that an empty waste pot is placed at the back of the MININEPHPLUS 7 5 3 Attach a new pipette tip on the end of the MININEPHPLUs hand held pipette 7 5 4 Check there is sufficient On Board buffer 1 SN107 in the drawer There needs to be at least 10mL Refer to the MININEPHPLUS User Guide for instructions on replenishing the buffer 7 5 5 The On Board Buffer 1 is automatically primed through after temperature T1 is selected and remains in the line to the hand held pipette 7 5 6 Switch on the analyser and printer if attached 7 5 7 Enter chemistry number Enter the chemistry number IgG Low Level 3 and press enter 7 5 8 Swipe chemistry card This message will only be displayed if this chemistry has never been used before or when changing antiser
12. um lot number Pass the swipecard through the swipe card reader moving in a left to right direction across the front of the analyser The magnetic stripe should be facing upwards 7 5 9 Check reagent lot number Press enter The MININEPHPLUS will automatically prime and select temperature T1 when the correct lot number is selected 7 5 10 IgG Low Level lot xxxx OK 1 Y 2 N Compare the details displayed with those on the antiserum label If the lot number displayed is identical to that printed on the antiserum vial select YES press 1 and continue to step 7 5 11 If the vial lot number is different from that displayed select NO press 2 and return to step 7 5 8 to allow the details of the correct batch to be entered 7 5 11 PipetteY N Block Y N Post priming there is a short period when the MININEPHPLUS stabilises its temperature Insert Code ZINO04 L Version 19 September 2011 Page 1 of 2 7 5 12 Prepare one MININEPH cuvette for each sample to be assayed Using the forceps provided with the MININEPHPLUS place a stirring bar in each cuvette and then using a pipette add 40uL of sample carefully to the bottom of each cuvette 7 5 13 Enter sample ID Enter an identity code e g 1 for the first sample to be assayed then press enter to continue refer to user manual for choice of identity codes Sample dilution 1 1 undiluted Accept the recommended dilution by pressing enter or type in a new dilution factor if an alternative dilution is to
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