TGS TA Rubella IgG Avidity
Contents
1. new DATA DISK was compiled Only the DATA DISK with the highest lot 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 IDS iSYS Cuvette Cube Pack of 960 cuvettes IDS iSYS System Liquid 1 bottle containing 5 liters of ready to use solution IDS iSYS Wash Solution 1 bottle containing 10 liters of ready to use solution IDS iSYS Trigger Set 7 1 250 mL bottle of Trigger A pre trigger solution 1 250 mL bottle of Trigger B trigger solution TGS TA D SORB Solution Pack of 2 bottles containing 1 liter of ready to use solution IDS iSYS Cartridge Checking System TGS TA ImmunoCleaner 6 bottles each containing 27 mL TGS TA Top Cap Set 300 red top caps to close the calibrator containers after first use Cod No TGS00001 Code No Code No Code No Code No Code No Code No Code No Code No IS CC1000 IS CS1000 IS CW 1000 IS CT1000 YC500009 IS 601000 YC500012 YC500010 IFU051TGS TA Version 01_EN 2 October 2014 Page 4 of 13 TGS TA Rubella IgG Avidity REF YB500047 manufactured by IDS France SAS 42 rue St phane Mazeau 21320 Pouilly en Auxois France and distributed by Technogenetics Srl manufactured by da IDS SA 101 103 rue Ernest Solvay 4000 Li ge Belgiqu2. 7 YB500047 TGS TA Rubella IgG Avidity RE REF YB500047 TGS TA Rubella IgG Avidity Ag INSTRUCTIONS e FOR USE a V EN English 50 03959 INTENDED USE The TGS TA Rubella IgG Avidity test is a chemiluminescent immunoassay CLIA for determination using TGS TA Analyser or IDS iSYS Multi Discipline Automated System of Avidity Index of specific IgG class antibodies directed against Rubella virus in samples of human serum or plasma EDTA Sodium Citrate The possibility of differentiating antibodies at high avidity from low avidity allows to supply interesting clinical information This assay is used as a diagnostic aid when assessing immunity status of patients related to Rubella virus infection 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 Rubella is a mild exanthematous and moderately contagious disease caused by Rubella virus which is the sole member of the Rubivirus genus of the Togavirus family The only known hosts for Rubella virus are humans and only one serotype has been identified Rubella is commonly called German measles or 3 day measles and it is the third of the six exanthemas of
3. 20 uL of Magnetic Particles 100 uL of Sample Diluent and 10 uL of sample or control for the calibrators the positive serum is supplied prediluted with Sample Diluent and the volume aspirated is 110 uL 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 a Only in the second sample cuvette b 200 uL of Buffer Avidity are added and the cuvette is incubated in the rotor at 37 C for 10 minutes b After this phase of incubation the magnetic particles are separated and washed 200 uL of conjugate are dispensed into the cuvette The reaction cuvette is incubated in the rotor at 37 C for 10 minutes 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 bonded to the solid phase expressed in RLU is directly proportional to the concentration of anti Rubella IgG present in the sample 8 The readings obtained are interpolated on the calibration curve and transformed into concentrations and Avidity Index IFU051TGS TA Version 01_EN 2 October 2014 Page 8 of 13 TGS TA Rubella IgG Avidity REF YB500047 QUALITY CONTROL To ensure the validity of the assay control sera at differing levels of avidity at least one low moderate av
4. surfactant and sodium azide lt 0 1 as preservative Sample Diluent Solution Phosphate Buffer containing bovine serum albumin a surfactant an inert blue colouring agent Pro Clin 300 and Gentamicin SO as preservatives Buffer Solution Borate Buffer containing a dissociating agent and sodium azide lt 0 1 as preservative Human serum with low Avidity of anti Rubella IgG antibodies in Phosphate Buffer containing bovine serum albumin a surfactant an inert blue colouring agent Pro Clin 300 and Gentamicin SO as preservatives Human serum with high Avidity of anti Rubella IgG antibodies in Phosphate Buffer containing bovine serum albumin a surfactant an inert blue colouring agent Pro Clin 300 and Gentamicin SO as preservatives IFU051TGS TA Version 01_EN 2 October 2014 Page 3 of 13 TGS TA Rubella IgG Avidity REF YB500047 All reagents are ready for use Reagents 1 2 3 and 4 are assembled in a single reagents cartridge unit The Calibrator concentrations are expressed in IU mL International Units and calibrated against WHO 1st IS RUBI 1 94 1996 The concentration settings specific for each production lot are recorded on the DATA DISK included in the kit DATA DISK A Mini CD containing data regarding all the products in the TGS TA line Reagents Calibrators Control Sera updated to the last production lot with the exclusion of products that have expired at the date when the
5. 1TGS TA Version 01_EN 2 October 2014 Page 7 of 13 TGS TA Rubella IgG Avidity REF YB500047 Calibration The TGS TA Analyzer instrument uses a memorised calibration curve master curve generated by the manufacturer for each lot 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 database Calibrators A and B are used to recalibrate the master curve in both for the instrument used and for the reagents on board To recalibrate analyse three replicates of the two calibrators A and B and one replicate of each control The concentration obtained with the controls make it possible to validate the new calibration Once recalibration of the master curve has been accepted and stored in memory all subsequent samples can be analysed without any further calibration except in the following cases when a reagent cartridge with a new lot number is loaded into the instrument when the controls do not fall within the range of acceptability after instrument maintenance when the validity of the recalibrated master curve has expired The validity of the recalibrated master curve for the TGS TA Rubella IgG Avidity kit is 21 days Recalibration management is handled automatically by the analyser Assay Press the start button 1 The system aspirates 100 uL of Sample Diluent
6. 5 samples from patients infected by Rubella of which 14 follow up samples coming from 2 patients in seroconversion and 4 patients with primary infection 5 samples from as many patients with primary infection within 1 month 2 samples with not dated primary infection and 74 samples from as many patients with not recent infection IFU051TGS TA Version 01_EN 2 October 2014 Page 10 of 13 TGS TA Rubella IgG Avidity REF YB500047 Following the interpretation given above it was possible to work out the diagnostic specificity and sensitivity relating to each group under evaluation Of 98 not immune patients 97 had IgG values below cut off by using methods TGS TA Rubella IgG The diagnostic specificity of method for IgG related to non immune patients was 99 0 95 Confidence Interval 93 6 99 9 For IgM the specificity calculated on 172 samples 98 not immune samples and 74 previous infection samples was 99 4 171 172 95 Confidence Interval 96 3 100 0 Among 95 samples of patients diagnosed as infected by Rubella in seroconversion monitoring and follow up the method TGS TA Rubella IgG has shown 93 positive samples so the sensitivity was 97 9 93 95 95 Confidence Interval 96 0 100 0 Among 21 samples from patients diagnosed as infected by Rubella in act the method TGS TA Rubella IgM has shown 20 positive samples and 1 negative sample so the diagnostic sensitivity was 95 2 20 21 95 Confidence Interv
7. 99 5 75 77 The relative agreement in low avidity samples was found to be 88 9 95 Confidence Interval 63 9 98 1 16 18 The 2 discordant samples have shown high avidity The relative agreement in high avidity samples was found to be 100 0 95 Confidence Interval 92 4 100 0 59 59 IFU051TGS TA Version 01_EN 2 October 2014 Page 12 of 13 TGS TA Rubella IgG Avidity REF YB500047 BIBLIOGRAPHY 10 11 12 13 14 DM Horstmann Rubella in Evans A S ed Viral infections of humans epidemiology and control New York Plenum Press 1976 409 427 Askin DF Intrauterine infections Neonatal Netw 2004 23 5 23 30 Cooper LZ Alford Jr CA Rubella In Remington JS Klein JO editors Infectious Diseases of the Fetus and Newborn infant Philadelphia PA Elsevier Saunders 2006 893 926 Atreya CD Mohan KV Kulkami S Rubella virus and birth defects molecular insights into the viral teratogenesis at the cellular level A Clin Mol Terato 2004 70 7 431 7 Department of Vaccines and Biologicals WHO Report of a meeting on preventing congenital Rubella syndrome immunization strategies surveillance needs Geneva World Health Organization 2000 Best JM Banatvala JE Rubella In AJ Zuckerman JE Banatvala JR Pattison eds Principles and practice of clinical virology 4 ed Chichester John Wiley 2000 387 418 Best JM O Shea S Tipples G Davies N Al Khusai
8. ISK 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 instructions for use provided The Avidity Index of IgG anti Rubella antibody in the TGS TA controls are recorded 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 According to the concentration of Rubella IgG previously tested 2 protocols are available They can be selected in analyte list parameters according to the following scheme Rubella IgG concentration between 10 and 300 IU mL Rubella Avidity IgG 10 300 Rubella IgG concentration gt 300 IU mL Rubella Avidity IgG gt 300 IFU05
9. TGS TA Rubella IgG Avidity REF YB500047 The reproducibility was calculated by analyzing the results of the determination of five sera at low and high avidity of anti Rubella IgG performed in 17 different sessions with three lots of reagents The table shows the results obtained with the 5 sera Sample Mean Index SD Index CV 1 0 14 0 01 7 1 2 0 72 0 03 4 2 3 0 68 0 04 5 9 4 0 69 0 05 7 2 5 0 72 0 03 4 2 Analytical Specificity Interferences A study based on the guidelines given in the CLSI document 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 Baia Intertennig Maximum tested concentration ubstances Free bilirubin 20 mg dL Conjugated bilirubin 20 mg dL Haemoglobin 1000 mg dL Triglycerides 3000 mg dL Use of lipaemic haemolysed and turbid samples is not in any case recommended Relative Sensitivity and Specificity The avidity index of IgG anti Rubella antibodies was determined using the TGS TA Rubella IgG Avidity kit and an immunoenzymatic assay available on the market in 77 samples 18 at low avidity and 59 at high avidity 2 samples gave rise to discordant results between the TGS TA assay and the other available method The global relative agreement was therefore found to be 97 4 95 Confidence Interval 90 1
10. al 74 1 99 8 By using TGS TA Rubella IgG Avidity kit in 95 samples from patients diagnosed as above the Avidity index of Rubella IgG in all 14 samples in seroconversion and followed samples with primary infection was low until 2 months from the onset of symptoms as well as in all 5 samples with infection in the last month and in 2 samples with recent infection without dating In all 74 samples with past infection the Avidity index of Rubella IgG was high 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 Rubella IgG Avidity kit have been assessed using a protocol based on the guidelines given in Clinical and Laboratory Standards CLSI document EP5 A2 The precision was calculated by analysing the results of 20 replicates of three sera at low and high index of anti Rubella IgG Avidity performed with two different lots of reagents in the same test run The table shows the results obtained with the 3 sera Sample brea Mean Index SD Index CV E eae ae E a oe a a 3 0 16 0 01 6 3 Py eee ee ee 004 3 0 65 0 09 13 8 Pe oe Em eee a a ee ee on 3 0 65 0 07 10 8 IFU051TGS TA Version 01_EN 2 October 2014 Page 11 of 13
11. artridge 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 The assay 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 assay is performed after more than 8 hours separate the serum or the plasma from the clot from the red blood cells and transfer the supernatant from the gel separating tubes to the secondary tubes Before being analysed samples may be kept in a refrigerator at 2 8 C for a maximum of 7 days If the assay is to be performed after more than 7 days store the samples frozen lt 20 C Avoid repeated freezing and thawing OPERATING PROCEDURE Carefully follow the instructions given in the user manual of the instrument to obtain reliable analytical results Loading of reagents All the reagents supplied in the kit are ready for use Before inserting the reagent cartridge in the system the magnetic particle container must be horizontally agitated by rotation in order to ensure correct resuspension of the particles Avoid generating foam when performing this operation Place the reagent cartridge in the reagent area of the instrument u
12. by SM Krause A Hesketh LM Jin L Enders G Interpretation of Rubella serology in pregnancy pitfalls and problems BMJ 2002 325 7356 147 148 Banatvala JE and Brown DWG 2004 Rubella Lancet 363 1127 1139 Banatvala JE Best JM O Shea S Dudgeon JA Persistence of Rubella antibodies following vaccination detection of viremia following experimental challenge Rev Infect Dis 1985 7 suppl 1 S 86 90 Thomas HIJ Morgan Capner P Roberts A Hesketh L Persistent Rubella specific IgM reactivity in the absence of recent primary Rubella and Rubella reinfection J Med Virol 1992 36 188 92 D O White and F J Fenner 1994 Medical Virology 4 ed Academic Press San Diego CA R Hamkar S Jalivand T Mokhtari Azad K Nouri Jelyani H Dahi Far H Soleimanjahi and R Nategh 2005 Assessment of IgM enzyme immunoassay and IgG avidity assay for distinguish between primary and secondary immune response to Rubella vaccine J Virol Methods 130 59 65 J Hoffman and U G Liebert 2005 Significance of avidity and immunoblot analysis for Rubella IgM positive serum samples in pregnant women J Virol Methods 130 66 71 V Herne K Hedman and P Reedik 1997 Immunoglobulin avidity in the serodiagnosis of congenital Rubella syndrome Eur J Clin Microbiol Infect Dis 16 763 766 TECHNOGENETICS S r l Via Vanvitelli 4 20129 Milano Italy IFU051TGS TA Version 01_EN 2 October 2014 Page 13 of 13
13. chilhood The disease is transmitted human to human via airborne droplet emission from the upper respiratory tract of active patients The incubation is about 2 to 3 weeks This disease is often mild and many patients are asymptomatic The primary symptoms are appearance of a rash on the face which spreads to the trunk and limbs and usually fades after three days low grade fever swollen glands joint pains neadache and conjunctivitis The severe syndromes happen when pregnant women are infected with Rubella virus especially if the infection happens in the first trimester of the pregnancy The infection can be transmitted to the fetus through placenta and the so called congenital Rubella syndrome CRS may occur which lead to miscarriage prematurity or stillbirth If the fetus survives the CRS serious defects may occur including blindness deafness or life threatening organ disorders Although rare in many industrialized countries because of the success of vaccination programs Rubella continues to occur where uptake of the vaccine is slow and in many developing countries with no vaccination IFU051TGS TA Version 01_EN 2 October 2014 Page 1 of 13 TGS TA Rubella IgG Avidity REF YB500047 program The World Health Organization WHO has therefore encouraged all countries to assess their Rubella immunostatus and introduce immunization and surveillance if appropriate Clinical diagnosis of Rubella
14. e and distributed by Technogenetics Srl Other Recommended Reagents TGS TA Rubella Avidity Control Set Cod No YB500048 Three 1 0 mL vials of Low moderate Avidity serum and three 1 0 mL vials of High Avidity of IgG anti Rubella antibodies WARNINGS AND PRECAUTIONS The reagents supplied in the TGS TA Rubella IgG Avidity 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 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 Dispo
15. he concentration of specific antibodies present in the sample in the calibrators and in the controls The ratio between the concentration of antibodies of the second cuvette b treated sample IgG not removed and the concentration of antibodies of the first cuvette a not treated sample Total IgG is the Index of Avidity of the Antibodies IgG Anti Rubella present in the sample IFU051TGS TA Version 01_EN 2 October 2014 Page 2 of 13 TGS TA Rubella IgG Avidity REF YB500047 AUTOMATION The TGS TA Analyser instrument automatically performs all the operations envisaged by the assay protocol addition of samples calibrators controls magnetic particles conjugates and chemiluminescence activation solutions to the reaction cuvette magnetic separation and washing of particles measurement of the emitted light The system calculates the assay 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 assay and to the patient MATERIALS AND REAGENTS Materials and reagents supplied Magnetic particles coated with inactivated Rubella antigen strain HPV 77 in Phosphate Buffer containing stabilising proteins Pro Clin 300 and sodium azide lt 0 1 as preservatives Mouse Monoclonal anti human IgG antibody labelled with an acridinium ester derivative conjugate in Phosphate Buffer containing stabilising proteins
16. idity serum and one high avidity serum must be measured every day in which dosage is performed If individual laboratory practice so dictates more frequent or more numerous controls may be performed for verification of assay results Follow local quality control procedures If TGS TA control sera are used the expected average concentration and the acceptability limits are those given on the DATA DISK included in the control set pack too If different control sera are used before using them the values expected with TGS TA reagents and system must be defined If the control values does not fall within the specified range of acceptability the related assay results are not valid and the respective samples must be analysed again In this case before repeating the tests a recalibration procedure must be performed CALCULATION AND INTERPRETATION OF THE RESULTS Calculation of the results The concentration of the anti Rubella IgG present in the samples that are being tested is automatically calculated by the system The results can be viewed on the display or printed The concentrations are expressed in IU mL Calculation of the analyte concentration in the sample takes place by interpolating the response obtained for each sample on a calibration curve calculated in accordance with a 4 parameter logistic fitting 4PL Y weighted periodically corrected according to the responses obtained for calibrators assay results For detailed informat
17. ion on how the system calculates the results please see the analyser user manual The measurement range of TGS TA Rubella IgG is 0 0 468 IU mL Values lower than 0 0 IU mL are extrapolated values the message OMR and or ORA appears and they are shown as equal to 0 0 IU mL Values higher than 468 IU mL are accompanied by the message OMR and or ORA and may retested after suitable dilution The Avidity elaborated by the instrument is expressed in Index The results of the samples may be interpreted in the following way Index Interpretation lt 0 20 The sample should be considered at Low Avidity IgG anti Rubella 0 20 0 25 The sample should be considered at Moderate Avidity IgG anti Rubella gt 0 25 The sample should be considered at High Avidity IgG anti Rubella IFU051TGS TA Version 01_EN 2 October 2014 Page 9 of 13 TGS TA Rubella IgG Avidity REF YB500047 The values reported above are indicative only Each laboratory will establish its own reference intervals LIMITS TO THE ASSAY METHOD For diagnostic purposes the results obtained with the TGS TA Rubella IgG Avidity 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 Heterophilic antibodies present in human serum samples may react with immunogl
18. is unreliable as Rubella virus infection can be asymptomatic in up to 50 of infected patients Furthermore the rash caused by Rubella virus and the accompanying symptoms are so similar to other viral infections that it is impossible for a physician to make a confirmed diagnosis on visual examination alone Thus serological tests are needed for a diagnosis especially when a patient is pregnant Rubella antibody testing are so the main diagnostic tool available to either establish a previous exposure to Rubella or Rubella vaccination or a current infection re infection with Rubella A primary acute Rubella infection is associated with an elevated IgM antibody response to the Rubella virus Rubella virus specific IgM are detectable after the incubation period and usually disappear after approximately 8 weeks although in rare cases they may be present a year or more after natural infection or vaccination and after asymptomatic reinfection 910 Rubella virus specific IgGs appear immediately after IgMs and they show a greater than fourfold rise in their titer and persist for life In addition to IgM and IgG determinations immunoglobulin G avidity testing has been shown to be useful for differentiating recent from past Rubella infection investigating suspected Rubella in pregnant women and congenital Rubella syndrome PRINCIPLE OF THE METHOD The TGS TA Rubella IgG Avidity kit for determination of the avidity index of specific anti R
19. obulin based reagents causing interference with in vitro immunological dosages Such samples may give rise to anomalous readings if analysed with the TGS TA Rubella IgG Avidity kit DIAGNOSTIC SPECIFICITY AND SENSITIVITY By the use of TGS TA Rubella IgG TGS TA Rubella IgM and TGS TA Rubella IgG Avidity kits it is possible to evaluate the patient immune status related to Rubella infection In the following table are reported the serological profiles of the 3 tests Rubella IgG Rubella IgM and Rubella IgG Avidity and their relative interpretation with respect to the virus exposure IgG anti IgM anti Ai g g IgG anti Rubella Avidity Interpretation Rubella Rubella 2nd level test Not Immune Individual susceptible negative negative TE o infection negative positive Suspected acute primary infection Fi 7 Recent primary infection last 2 positive positive Low Avidity l months from sample collection Recent previous infection with positive positive High Avidity persistent IgM presence more than last 2 months from sample collection positive negative High Avidity Past infection Using the TGS TA Rubella IgG TGS TA Rubella IgM TGS TA Rubella IgG Avidity tests the diagnostic specificity and sensitivity were assessed in the following population of selected samples examined with different methods and classified according to the rule of general consensus 98 not immune patients 9
20. se 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 IFU051TGS TA Version 01_EN 2 October 2014 Page 5 of 13 TGS TA Rubella IgG Avidity REF YB500047 Operating precautions Reliable results can only be obtained by strictly complying with these instructions and carefully 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 Analyzer system The components of the reagent cartridge must not be removed from the cartridge and reassembled Do not use the kit after its expiry date REAGENT PREPARATION 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 until the expiry date After opening the reagent c
21. sing the rack provided and leave it to be agitated for at 1 hour before use IFU051TGS TA Version 01_EN 2 October 2014 Page 6 of 13 TGS TA Rubella IgG Avidity REF YB500047 Positioning of the reagent cartridge simultaneously determines reading of the identification 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 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 IgG anti Rubella antibody concentration in the calibrators are recorded in the DATA D
22. ubella IgG employs an indirect two step immunological method based on the principle of chemiluminescence The test can be done only by using samples previously tested to detect the presence of IgG anti Rubella The specific antigen is used to coat the magnetic particles solid phase and an anti human IgG antibody is labelled with an acridinium ester derivative conjugate Each specimen is dispensed into two adjoining cuvettes cuvette a as reference and b in which a buffer able to prevalently break the binding Antigen Ag Antibody Ab if the antibody is at low avidity is added During initial incubation the specific antibodies present in the sample in the calibrators or in the controls bond with the solid phase In the second cuvette b only at the end of the first incubation a second incubation is performed in a buffer able to break the binding between the Ag of the magnetic particles and the Antibodies IgG of the samples if they are present at low avidity During the last incubation the conjugate reacts with the anti Rubella IgG antibodies captured 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 labelled conjugate that remains bound to the solid phase is assessed by activation of the chemiluminescence reaction and measurement of the light signal The generated signal expressed in relative light units RLU is indicative of t
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