TGS TA LKM-1 - Technogenetics
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1. TGS TA LKM 1 REF YA500232 REF YA500232 M TEC A TGS TA TGS TA LKM 1 R GENETIC INSTRUCTIONS FOR USE 8 C eo 50 EN English INTENDED USE The TGS TA LKM 1 Liver Kidney Microsomes type 1 test is a chemiluminescent immunoassay CLIA used for quantitative determination using the TGS TA Analyzer or IDS iSYS Multi Discipline Automated System of specific IgG class antibodies directed against Liver Kidney Microsomes type 1 in samples of human serum or plasma EDTA Sodium Citrate This dosage is used as a diagnostic aid when assessing type 2 Autoimmune Hepatitis EAI or AIH infections This product must be used in strict compliance with the instructions given in this document by professional users CAUTION Medical decisions must not be based exclusively on the result of this test but must take into account all available clinical and laboratory data as a whole CLINICAL SIGNIFICANCE Autoimmune Hepatitis AIH is a chronic inflammation of the liver of unknown origin in which immune tolerance to hepatocytes is lost The disease is detected by the presence of a number of characteristic alterations from the histological point of view when blood tests show the presence of gamma globulins and antibodies The disease is found mainly in females both of paediatric and adult ages It is an autoimmune process and therefore is associated sometimes with othe2. periodically corrected according to the responses obtained for dosage of the calibrators For detailed information on how the system calculates the results please see the operating manual for the system Interpretation of the results The dosage range measurable for TGS TA LKM 7 is 0 0 300 AU mL Readings lower than 0 0 AU mL are extrapolated values the message OMR and or ORA appears and they are shown as equal to 0 0 AU mL Readings higher than 300 AU mL are accompanied by the message OMR and or ORA and may be retested after suitable dilution The results of the samples may be interpreted in the following way AU mL Interpretation lt 15 The sample must be considered to be negative for the presence of anti LKM 1 antibodies 2 15 The sample must be considered to be positive for the presence of anti LKM 1 antibodies The above readings are to be considered only suggested readings Each laboratory must establish its own reference ranges LIMITS OF THE ASSAY METHOD For diagnostic purposes the results obtained with the TGS TA LKM 1 kit and the TGS TA Analyser system must be used together with the other clinical and laboratory data available to the physician Bacterial contamination of the sample and heat inactivation may influence the result of the dosage Heterophyllous antibodies present in human serum samples may react with immunoglobulin based reagents causing interference with in vitro immuno
3. Barut A Frank H and Manns MP Epitope mapping of cytochrome P450 2D6 autoantigen in patients with chronic hepatitis C during alpha interferon treatment J Hepatol 30 3 366 375 1999 IFUO60TGS TA Version 01_EN 15 January 2015 Page 13 of 14 TGS TA LKM 1 REF YA500232 12 Miyakawa H Kikazawa E Abe K Kikuchi K Fujikawa H Matsushita M Kawaguchi N Morizane T Ohya K Kako M Detection of anti LKM 1 anti CYP2D6 by enzyme linked immunosorbent assay in adult patients with chronic liver diseases Autoimmunity 1999 30 2 107 14 13 Kekar N Ma Y Davies ET Cheeseman P Mieli Vergani G Vergani D Detection of liver kidney microsomal type 1 antibody using molecularly based immunoassay Journal of Clinical Pathology 2002 55 12 906 909 TECHNOGENETICS S r l Via Vanvitelli 4 20129 Milano Italy IFUO60TGS TA Version 01_EN 15 January 2015 Page 14 of 14
4. HCV hepatitis even though the epitopes recognised in HCV patients are different from those recognised in patients suffering from type 2 AIH Anti LKM 1 antibodies are found in around 90 of patients with type 2 AIH and can be identified by the IFA test using sections of mouse liver and kidney tissue It is also possible to test for anti LKM 1 antibodies with indirect solid phase assays using the recombinant protein CYP2D6 with high levels of sensitivity and specificity PRINCIPLE OF THE METHOD The TGS TA LKM 1 kit for quantitative determination of specific IgG class antibodies directed against Liver Kidney Microsomes employs an indirect two step immunological method based on the principle of chemiluminescence The specific antigen used to coat the magnetic particles solid phase and the G protein able to bond with human IgGs is marked with an acridinium ester derivative conjugate During initial incubation the specific antibodies present in the sample in the calibrators or in the controls bond with the solid phase During the second incubation the conjugate reacts with the anti LKM 1antibodies sequestered by the solid phase After each incubation the material that has not bound with the solid phase is removed by aspiration and subsequent washing The quantity of marked conjugate that remains bound to the solid phase is assessed by activation of the chemiluminescence reaction and measurement of the luminous signal The
5. ready for use Leave the calibrators at room temperature for 10 minutes and then gently shake the contents either manually or using a vortex avoiding the formation of foam When using the calibrators for the first time remove the guarantee seal and the white sealing cap before placing them in the analyser If the calibrators have already been used the container will have a top cap red cap with no guarantee seal Remove the red closing cap before placing them in the analyser Place the calibrators in the samples area of the analyser see the analyser user manual on how to identify them in the analyser Bar code data must be entered manually if the label is damaged or if it is unreadable The readings for the LKM 1 antibody concentration in the calibrators are recorded in the DATA DISK and automatically transferred to the analyser At the end of the session the calibrator containers must be closed with the top caps red caps provided and stored at 2 8 C until they are used again The calibrators can be used for a maximum of four times Loading of controls Place the controls in the samples area of the analyser See the analyser user manual on how to identify them in the analyser If there is no bar code on the control or if it is not readable the control identification data must be entered manually If TGS TA Controls are used see the usage instructions provided The readings for the LKM 1 antibody concentration in the TGS TA controls are re
6. autoimmune hepatitis AIH 1 118 from patients with primary biliary cirrhosis PBC 40 with hepatitis C HCV 19 with hepatitis B HBV 20 primary sclerosing cholangitis PSC and 160 normal samples 7 samples tested positive and 408 negative Diagnostic specificity 98 3 408 415 Of the 7 samples that proved to be not negative 4 belonged to the group of patients diagnosed with AIH 1 1 belonged to the group of patients diagnosed with HCV 1 belonged to patients diagnosed with HBV and 1 to the group of normal samples In the presumably positive population studied including 8 samples from patients diagnosed with type 2 autoimmune hepatitis AIH 2 8 proved to be positive Diagnostic sensitivity 100 0 8 8 Based on the diagnostic specificity and sensitivity results the diagnostic agreement is 98 3 416 423 PERFORMANCES Caution the data presented do not represent the operating specifications of the kit but serve as experimental proof of how the kit works within these specifications in the manner envisaged by the manufacturer Precision and Reproducibility The precision and the reproducibility of the TGS TA LKM 17 kit have been assessed using a protocol based on the guidelines given in Clinical and Laboratory Standards CLSI document EP5 A2 IFUO60TGS TA Version 01_EN 15 January 2015 Page 10 of 14 TGS TA LKM 1 REF YA500232 The precision was calculated by analysing the results o
7. bovine serum albumin a surfactant an inert blue colouring agent Pro Clin 300 and Gentamicin SO as preservatives Serum with low concentration of anti LKM 1 antibodies containing sodium azide lt 0 1 as preservative Serum with high concentration of anti LKM 1 antibodies containing sodium azide lt 0 1 as preservative All reagents are ready for use Reagents 1 2 3 and 4 are assembled in a single kit forming the reagent cartridge The Calibrator concentrations are expressed in AU mL arbitrary units and calibrated against an internal reference standard The concentration settings specific for each production batch are recorded on the DATA DISK included in the kit IFUO60TGS TA Version 01_EN 15 January 2015 Page 3 of 14 TGS TA LKM 1 REF YA500232 DATA DISK A Mini CD containing data regarding all the products in the TGS TA line Reagents Calibrators Control Serums updated to the last production batch with the exclusion of products that have expired at the date when the new DATA DISK was compiled Only the DATA DISK with the highest batch number needs to be kept to maintain the information required for correct operation of the system up to date Materials and reagents required but not supplied in the kit TGS TA Analyzer 120 Cod No TGS00001 TGS TA Cuvette Cube Code No IS CC1000 Pack of 960 cuvettes TGS TA System Liquid Code No IS CS1000 1 bottle containing 5 lite
8. conformity with the instructions supplied Safety precautions This product contains material of animal origin and therefore must be handled as if it contains infecting agents This product contains components of human origin All units of serum or plasma used to produce the reagents in this kit have been analysed with FDA approved methods and found not to be reactive due to presence of HBsAg anti HCV anti HIV1 and anti HIV2 However since no analysis method is able to guarantee the absence of pathogenic agents all material of human origin must be considered to be potentially infected and handled as such In the event of damaged packaging or accidental leakage decontaminate the area concerned with a diluted solution of sodium hypochlorite after putting on suitable personal protective equipment overall gloves goggles Dispose of the material use for the clean up and of the packaging involved in the leakage according to national regulations for disposal of potentially infected waste In the event of damaged packaging or accidental leakage do not use the reagents to perform the assay Some reagents contain sodium azide as a preservative Since sodium azide may react with lead copper and leaded brass forming explosive azides in piping it is recommended that reagents or waste are not poured down drains but are disposed of in compliance with the national regulations on disposal of potentially hazardous waste Operating precautions Reliab
9. corded in the DATA DISK and automatically transferred to the analyser Select the required parameters for each control Loading of samples Place the samples in the samples area of the analyser see the analyser user manual on how to identify them in the analyser If there is no bar code on the sample or if it is not readable the sample identification data must be entered manually Select the required parameters for each sample Calibration The TGS TA Analyser instrument uses a memorised calibration curve master curve generated by the manufacturer for each batch of reagent cartridges The master curve parameters together with the calibrator concentration settings are stored in the DATA DISK and transferred to the instrument s data base Calibrators A and B are used to recalibrate the master curve in both for the instrument used and for the reagents on board IFUO60TGS TA Version 01_EN 15 January 2015 Page 7 of 14 TGS TA LKM 1 REF YA500232 To perform recalibration analyse the two calibrators A and B in triplicate and the controls singly The concentration readings obtained with the controls make it possible to validate the new calibration Once recalibration of the master curve has been accepted and memorised all subsequent samples can be analysed without any further calibration except in the following cases when a reagent cartridge with a new batch number is loaded into the in
10. cument EP7 A2 has shown that the dosage performances are not influenced by the presence in the sample of the potentially interfering substances listed in the table below up to the tested concentration deca inter onng Maximum tested concentration ubstances Free bilirubin 20 mg dL Conjugated bilirubin 20 mg dL Haemoglobin 1000 mg dL Tryglicerides 3000 mg dL Use of lipaemic haemolysed and turbid samples is not in any case recommended Analytical Specificity Cross reactions In order to assess potential cross reactions of the antigen used to sensitise the microparticles a study was conducted using 40 samples all with high levels of other antibodies and negative for anti LKM IgG The samples used were subdivided as follows SS A 2 SS B 2 U1 snRNP 2 Jo 1 2 Scl 70 2 Cenp B 2 Sm 2 PR3 2 MPO 2 B2 GLI CL 2 Gliadin 2 t TG 2 CCP 2 GBM 2 dsDNA 2 TG 3 TPO 3 and Rheumatoid Factor RF 4 The study did not show any significant cross reaction to the antigen in the solid phase with the other antibodies Saturation effect at high doses Some immunological methods used to determine samples containing the analyte at extremely high concentrations may supply apparent levels of underestimated analyte Hook effect The method used in the TGS TA LKM 1 kit since it uses two incubations is not subject to this effect A sample with an extremely high concentration above the measurement
11. e referred to as the Limit of Detection LoD that is the smallest quantity of analyte that the method is able to measure the formula for calculating LoD LoB Cg SD in which LoB is the Limit of Blank SD is the estimated standard deviation of the distribution of the sample at low concentration and Cz is derived from 95 percentile of the standard Gaussian distribution was applied Three low concentration samples of analyte were used determined singly with one batch of reagents in 15 different tests The Limit of Detection of the TGS TA LKM 1 kit proved to be 3 6 AU mL IFUO60TGS TA Version 01_EN 15 January 2015 Page 11 of 14 TGS TA LKM 1 REF YA500232 In the other protocol calculation of the Minimum Detectable Concentration MDC is envisaged 20 replicates of the solution of the 0 AU mL Standard of the Master curve were used The average and the standard deviation DS were formulation on a batch of the kit The RLU value related to the average 2 6 DS was interpolated on the curve and the related concentration was obtained from this value The analytical sensitivity expressed as the Minimum Detectable Concentration is 0 7 AU mL The minimum detection values together with considerations of a clinical kind and the results of comparison with reference methods contributed to the definition of the cut off value Analytical Specificity Interferences A study based on the guidelines given in the CLSI do
12. epatitis what is the gold standard Hepatology 2010 51 1 350 1 3 Ngu JH et al Population based epidemiology study of autoimmune disease a disease of older women J Gastroenterol Hepatol 2010 25 10 1681 6 4 Navaneethan U and Shen B Hepatopancreatobiliary manifestations and complications associated with inflammatory bowel disease Inflamm Bowel Dis 2010 16 9 1598 619 5 Nakamura H et al Prevalence of interrelated autoantibodies in thyroid diseases and autoimmune disorders J Endocrinol Invest 2008 31 10 861 5 6 Rizzetto M Swana G Doniach D Microsomal antibodies in active chronic hepatitis and other disorders Clin Exp Immunol 1973 15 331 44 7 Gueguen M Meunier Rotival M Bernard O Alvarez F Anti liver kidney microsome antibody recognizes a cytochrome P450 from IID subfamily J Exp Med 1988 168 801 6 8 Philipp T Durazzo M Trautwein C Alex B Straub P Lamb JG et al Recognition of uridine diphosphate glucoronisyl transferases by LKM 3 antibodies in chronic hepatitis D Lancet 1994 344 578 81 9 Lenzi M Ballardini G Fusconi M Cassani F Selleri L Volta U et al Type 2 autoimmune hepatitis and hepatitis C virus infection Lancet 1990 335 258 9 10 Declos Valee JC Hajaoui O Yamamoto AM Jacqz Aigrin E and Alvarez F Conformational epitopes on CYP2D6 are recognized by Liver kidney microsomal antibodies Gastroenterology 108 467 470 1995 11 Dalekos GN Wedemeyer H Obermayer Straub P Kayser A
13. f 20 replicates of four serums one negative and three positive with differing concentrations of LKM 1 antibodies performed with two different batches of reagents in the same test session The concentration of the negative anti LKM 1 serum N1 was always 0 0 AU mL with reagent batch no 1 and within the range from 0 to 0 7 AU mL with batch no 2 The table shows the results obtained with the 3 positive serums Reagents Average Concentration SD sample Batch fo 3 AU mL AU mL ye rr en ee ene 65 6 e 26S l 40 2 61 8 2 47 4 0 E A lt sesusmceemes ALe O BBB A 2 162 0 6 00 3 7 Le eee A AA A Haman UME canard ceases Ot occsteceeetace 3 2 198 9 11 51 5 8 The reproducibility was calculated by analysing the results of the determination of five serums one negative and four positive with differing concentrations of anti LKM 1 performed singly in 30 different sessions with two different batches of reagents The concentration of the negative anti LKM 1 serum N1 was always within the range from 0 0 and 0 9 AU mL The table shows the results obtained with the 4 positive serums Sample TARR Te i ents CV 1 58 9 3 16 5 4 2 149 2 8 89 6 0 3 192 8 13 62 7 1 4 86 8 4 47 5 1 Analytical Sensitivity The analytical sensitivity of the TGS TA LKM 1 kit was assessed using a protocol based on the guidelines given in Clinical and Laboratory Standards CLSI document EP17 A In one cas
14. generated signal expressed in relative light units RLU is indicative of the concentration of specific antibodies present in the sample in the calibrators and in the controls AUTOMATION The TGS TA Analyser instrument automatically performs all the operations envisaged by the dosage protocol addition of samples calibrators controls magnetic particles conjugates and chemiluminescence activation solutions to the reaction container magnetic separation and washing of particles measurement of the light emitted The system calculates the dosage results for the samples and controls by means of a stored calibration curve and prints a report that includes all the information related to the dosage and to the patient IFUO60TGS TA Version 01_EN 15 January 2015 Page 2 of 14 TGS TA LKM 1 REF YA500232 MATERIALS AND REAGENTS Materials and reagents supplied Magnetic particles coated with LKM 1 cytochrome P450 2D6 recombinant antigen in Phosphate Buffer containing stabilising proteins surfactant Pro Clin 300 and sodium azide lt 0 1 as preservatives Conjugated Diluent Solution Phosphate Buffer containing stabilising proteins surfactant and sodium azide lt 0 1 as preservative G protein conjugated with an acrinidium ester derivative conjugate in Phosphate Buffer containing surfactant and sodium azide lt 0 1 as preservative Sample Diluent Solution Phosphate Buffer containing
15. le results can only be obtained by strictly complying with these instructions and scrupulously following what is written in the operating manual for the instrument The reagents supplied in the kit must be used only with the TGS TA Analyser system IFUO60TGS TA Version 01_EN 15 January 2015 Page 5 of 14 TGS TA LKM 1 REF YA500232 The components of the reagent cartridge must not be removed from the cartridge and reassembled Do not use the kit after its use by date PREPARATION OF THE REAGENTS The reagents supplied in the kit are all ready for use REAGENT STORAGE AND STABILITY Store the reagents supplied in the kit at 2 8 C in a vertical position in a dark place In these conditions unopened reagent cartridge and calibrators are stable up to the use by date After opening the reagent cartridge can be used for 60 days if kept in a refrigerator at 2 8 C or in the analyser After opening the calibrators can be used for 60 days if kept in a refrigerator at 2 8 C and if they have not been left in the analyser for more than 6 hours per session Do not freeze the reagents and calibrators SAMPLE PREPARATION AND STORAGE Dosage must be performed on samples of human serum and plasma EDTA Sodium Citrate Use of lipaemic haemolysed and turbid samples is not recommended If the dosage is performed after more than 8 hours separate the serum or the plasma from the clot from the red globules and f
16. logical dosages Such samples may give rise to anomalous readings if analysed with the TGS TA LKM 1 kit IFUD60TGS TA Version 01_EN 15 January 2015 Page 9 of 14 TGS TA LKM 1 REF YA500232 EXPECTED VALUES The samples of 100 individuals selected randomly from the normal routine work of the laboratory were analysed to check the presence of anti LKM 1 antibodies All samples analysed proved negative with an average reading of 0 9 AU mL and a standard deviation of 5 6 AU mL Using the results obtained the Limit of Blank LoB the highest reading that can be expected in a series of samples that do not contain the analyte was calculated The Limit of Blank determined as 95 percentile of the negative population proved equal to 1 6 AU mL with the Reagent batch no 2 DIAGNOSTIC SENSITIVITY AND SPECIFICITY CLINICAL A total of 423 samples were tested with the TGS TA LKM 1 kit 8 of the samples were from patients suffering from type 2 autoimmune hepatitis AIH 2 58 from patients suffering from type 1 autoimmune hepatitis AIH 1 118 from patients with primary biliary cirrhosis PBC 40 from patients with hepatitis C HCV 19 from patients with hepatitis B HBV 20 samples from patients with primary sclerosing cholangitis PSC while 160 samples were presumed normal originating from routine laboratory testing In the presumably negative population studied including 58 samples from type 1
17. r autoimmune disorders such as ulcerative colitis and Graves Disease Autoimmune Hepatitis is classified according to the specific autoantibodies detected which are for type 1 principally represented by ANA anti nuclear and SMA anti smooth muscle antibodies type 2 on the contrary is typified by LKM 1 type 1 liver kidney microsomal antibodies type 3 is characterised by anti SLA soluble liver antigen and anti LP liver pancreas antigen antibodies The presence of anti LKM autoantibodies was described for the first time in 1973 It was subsequently demonstrated that these are a heterogeneous family of autoantibodies and depending on the target antigen are classified in three types known as anti LKM 1 2 and 3 Cytochrome P4502D6 was identified as the autoantigen of the anti LKM 1 antibodies Cytochrome P4502C9 and UDP glucuronosyltransferase were later identified respectively as the targets of the anti LKM 2 and anti LKM 3 antibodies While anti LKM 2 IFUO60TGS TA Version 01_EN 15 January 2015 Page 1 of 14 TGS TA LKM 1 REF YA500232 antibodies have been associated with drug induced hepatitis anti LKM 1 antibodies have been associated with type 2 AIH and are a characteristic marker for this disease not usually associated with the presence of ANA and or ASMA of type 1 AIH However anti LKM 1 antibodies are not specific to type 2 AIH in that they may be found in 5 10 of the patients with chronic
18. range of anti LKM 1 antibodies confirmed the absence of the hook effect up to the concentration of 22000 AU mL IFUO60TGS TA Version 01_EN 15 January 2015 Page 12 of 14 TGS TA LKM 1 REF YA500232 Relative Sensitivity and Specificity The presence of anti LKM 1 antibodies was determined using the TGS TA LKM 1 kit and an ELISA dosage method available on the market in 343 samples 8 of the samples were from patients suffering from type 2 autoimmune hepatitis AIH 2 118 from patients suffering from primary biliary cirrhosis PBC 58 from patients with type 1 autoimmune hepatitis AIH 1 40 from patients with hepatitis C HCV 19 from patients with hepatitis B HBV 20 samples from patients with primary sclerosing cholangitis PSC while 80 presumably normal samples were obtained from routine laboratory testing 8 samples gave rise to discordant results between the TGS TA dosage and the ELISA dosage available on the market The relative concordance was therefore found to be 97 7 95 Confidence Interval 95 3 98 9 335 343 The relative sensitivity was therefore found to be 100 0 95 Confidence Interval 51 7 98 5 6 6 The relative specificity was therefore found to be 97 6 95 Confidence Interval 95 2 98 9 329 337 BIBLIOGRAPHY 1 Strassburg CP Autoimmune hepatitis Best Pract Res Clin Gastroenterol 2010 24 5 667 82 2 Kachar R and Fallon M Diagnostic criteria for autoimmune h
19. rom the separating tubes with gel Before being analysed samples may be kept in a refrigerator at 2 8 C for a maximum of 7 days If the dosage is to be performed after more than 7 days store the samples frozen lt 20 C Avoid repeated freezing and thawing ASSAY PROCEDURE Scrupulously follow the instructions given in the operating manual of the instrument to obtain reliable analytical results Loading of reagents All the reagents supplied in the kit are all ready for use Before inserting the reagent cartridge in the system the magnetic particle container must be horizontally agitated by rotation in order to favour resuspension of the particles Avoid generating foam when performing this operation Place the reagent cartridge in the reagent area of the instrument using the guide provided and leave it to be agitated for at least 40 minutes before use IFUO60TGS TA Version 01_EN 15 January 2015 Page 6 of 14 TGS TA LKM 1 REF YA500232 Positioning of the reagent cartridge simultaneously determines reading of the identity bar code If the cartridge label is damaged or if it is not readable the reagent cartridge identification data can be entered manually The instrument automatically maintains the magnetic particles constantly agitated If the reagent cartridge is removed from the instrument store it at 2 8 C in a vertical position in a dark place Loading of calibrators TGS TA calibrators are
20. rs of ready to use solution TGS TA Wash Solution Code No IS CW1000 1 bottle containing 10 liters of ready to use solution TGS TA Trigger Set Code No IS CT1000 1 250 mL bottle of Trigger A pre trigger solution 1 250 mL bottle of Trigger B trigger solution TGS TA D SORB Solution Code No YC500009 Pack of 2 bottles containing 1 liter of ready to use solution TGS TA Cartridge Checking System Code No IS 601000 TGS TA Top Cap Set Code No YC500010 300 red top caps to close the calibrator containers after first use Manufactured by IDS France SAS 42 rue St phane Mazeau 21320 Pouilly en Auxois France and distributed by Technogenetics Srl Manufactured by IDS S A Rue E Solvay 101 B 4000 Li ge Belgium and distributed by Technogenetics Srl IFUO60TGS TA Version 01_EN 15 January 2015 Page 4 of 14 TGS TA LKM 1 REF YA500232 Other Recommended Reagents TGS TA LIVER CONTROL SET Code No YA500233 Three 1 0 mL vials of negative human serum and three 1 0 mL vials of human serum positive for anti LKM 1 antibodies WARNINGS AND PRECAUTIONS The reagents supplied in the TGS TA LKM 1 kit are only for in vitro diagnostic use and not for in vivo use in humans or animals This product must be used in strict compliance with the instructions given in this document by professional users Technogenetics cannot be held responsible for any losses or damages caused by use not in
21. strument when the control readings do not fall within the range of acceptability when the instrument maintenance procedure is performed when the validity of the recalibrated master curve has expired The validity of the recalibrated master curve for the TGS TA LKM 1 kit is 21 days Control of recalibration is performed automatically by the instrument Assay Press the start button 1 The system aspirates 80uL of Sample Diluent 40uL of Magnetic Particles 100uL of Sample Diluent and 10uL of the sample calibrators or control The aspirated solutions and suspension are dispensed into the reaction cuvette The reaction cuvette is incubated in the rotor at 37 C for 10 minutes After this phase of incubation the magnetic particles are separated and washed 40uL of conjugate and 160 uL of conjugate diluent are dispensed into the cuvette The reaction cuvette is incubated in the rotor at 37 C for 20 minutes Dap N After this last phase of incubation the magnetic particles are separated and washed and the cuvette is transferred to the reading chamber 7 The quantity of conjugate bound to the solid phase expressed in RLU is directly proportionate the concentration of LKM 1 antibodies present in the sample 8 The responses obtained are interpolated on the calibration curve and transformed into concentrations QUALITY CONTROL To ensure the validity of the dosage control serums at differing levels of concen
22. tration at least one negative serum and one positive serum must be measured every day in which dosage is performed If your laboratory requires a more frequent use or a higher number of controls to check the dosage results comply with the set quality control procedure If TGS TA control serums are used the expected average readings and the acceptability limits are those given on the DATA DISK included in the control pack too If different control serums are used before using them the readings expected with TGS TA reagents and system must be defined If the control reading does not fall within the specified range of acceptability the related dosage results are not valid and the respective samples must be analysed again In this case before repeating the dosage a recalibration procedure must be performed IFUO60TGS TA Version 01_EN 15 January 2015 Page 8 of 14 TGS TA LKM 1 REF YA500232 CALCULATION AND INTERPRETATION OF THE RESULTS Calculation of the results The concentration of the anti LKM 1 antibodies present in the samples that are being tested is automatically calculated by the system The readings can be viewed on the display or printed The concentrations are expressed in AU mL Calculation of the analyte concentration in the sample takes place by reading the response obtained for each sample on a calibration curve processed by a logistic fitting system with four parameters 4PL Y weighted
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