TGS TA TOXO IgG
Contents
1. 98 1 579 597 The relative sensitivity was found to be 94 7 95 Confidence Interval 91 3 96 8 284 300 The relative specificity was found to be 99 3 95 Confidence Interval 97 3 99 9 295 297 IFU043TGS TA Version 01_EN 10 July 2014 Page 14 of 15 TGS TA TOXO IgG REF YB500032 BIBLIOGRAPHY 1 Ryan KJ Ray CG editors 2004 Sherris Medical Microbiology 4 ed McGraw Hill pp 723 7 ISBN 0838585299 2 Jones JL et al Congenital Toxoplasmosis A Review CME Review Article Vol 56 Number 5 2001 296 305 3 Torda A 2001 Toxoplasmosis Are cats really the source Aust Fam Physician 30 8 743 7 PMID 11681144 4 Jones JL Kruszon Moran D Wilson M McQuillan G Navin T McAuley JB Toxoplasma gondii infection in the United States seroprevalence and risk factors Am J Epidemiol 2001 154 4 357 65 5 Guerina NG Hsu HW Meissner HC Maguire JH Lynfield R Stechenberg B et al Neonatal serologic screening and early treatment for congenital Toxoplasma gondii infection The New England Regional Toxoplasma Working Group N Engl J Med 1994 330 26 1858 63 Montoya JG Liesenfeld O 2004 Toxoplasmosis Lancet 363 1965 76 Montoya JG Laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis J Infect Dis 2002 185 Suppl 1 S73 82 8 Remington JS McLeod R Thulliez P Desmonts G 2001 Toxoplasmosis p 205 346 In J S Rem2. TGS TA TOXO IgG REF YB500032 REF YB500032 Fiaa TGS TA TOXO IgG ATRN INSTRUCTIONS FOR USE 8 C IVD xX Z IVD a Te EN English INTENDED USE The TGS TA TOXO lgG test is a chemiluminescent immunoassay CLIA for quantitative determination using TGS TA Analyser or IDS iSYS Multi Discipline Automated System of specific IgG class antibodies directed against the Toxoplasma gondii TOXO in samples of human serum or plasma EDTA Sodium Citrate This assay is used as a diagnostic aid when assessing immunity status of patients related to Toxoplasma 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 Toxoplasmosis is a parasitic disease caused by infection with the obligate intracellular protozoan Toxoplasma gondii The parasite infects most genera of warm blooded animals including humans but the primary host is the felid cat family Animals are infected by eating infected meat by ingestion of feces of a cat that has itself recently been infected and by transmission from mother to fetus Cats are the primary source of infection to human hosts although contact with raw meat especially pork is a more s
3. EP5 A2 The precision was calculated by analysing the results of 20 replicates of five sera one negative and four positive with differing concentrations of anti Toxoplasma gondii IgG performed with two different lots of reagents in the same test run The concentration of the negative anti Toxoplasma gondii IgG TXG N3 serum gave results always 0 0 IU mL and from 0 4 to 0 6 IU mL respectively with reagent lots nos 2 and 3 The table shows the results obtained with the 4 positive sera IFU043TGS TA Version 01_EN 10 July 2014 Page 11 of 15 TGS TA TOXO IgG REF YB500032 Sample fen a no fara GNA 2 3 4 The reproducibility was calculated by analysing the results of the determination of six sera one negative and five positive with differing concentrations of anti Toxoplasma gondii IgG performed in 17 different sessions with three lots of reagents The concentration of the negative anti Toxoplasma gondii IgG TXG N3 serum gave results in the range from 0 0 to 0 3 IU mL The table shows the results obtained with the 5 positive sera Sample aes nee arabs ON 3 1 5 0 0 28 5 6 2 77 0 36 4 7 3 13 9 0 83 6 0 4 29 9 2 26 7 6 5 54 1 5 61 10 4 Linearity of the Dilutions The linearity of the TGS TA TOXO IgG kit was assessed using a protocol based on the guidelines given in Clinical and Laboratory Standards CLSI document EP6 A Serial
4. The components of the reagent cartridge must not be removed from the cartridge and reassembled Do not use the kit after its expiry date IFU043TGS TA Version 01_EN 10 July 2014 Page 5 of 15 TGS TA TOXO IgG REF YB500032 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 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 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 f
5. accordance with a 4 parameter logistic fitting 4PL Y weighted periodically corrected according to the responses obtained for calibrators assay results For detailed information on how the system calculates the results please see the analyser user manual Interpretation of the results The measurement range of TGS TA TOXO IgG is 0 0 78 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 78 IU mL are accompanied by the message OMR and or ORA and may retested after suitable dilution The results of the samples may be interpreted in the following way IU mL Interpretation lt 1 5 The sample must be considered to be Negative for anti Toxoplasma gondii IgG 21 5 The sample must be considered to be Positive for anti Toxoplasma gondii IgG 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 TOXO IgG 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 immunoglobulin based reagents causing interference with in vitro
6. dilutions performed by using System Liquid of 3 sera with high concentrations of anti Toxoplasma gondii IgG were dosed The results of this study are summed up in the table below IFU043TGS TA Version 01_EN 10 July 2014 Page 12 of 15 TGS TA TOXO IgG REF YB500032 Measured Expected Sample Dilution factor concentration concentration nee IU mL IU mL ji 1 37 5 37 5 100 1 8 3 7 4 7 78 7 1 61 7 61 7 100 2 3 In any case it must be stressed that non all the sera when measured at differing dilutions are able to provide linear results with the measurable range since the result depends not only on the concentration but also on the affinity of the antibodies present in the sample Analytical Sensitivity The analytical sensitivity of the TGS TA TOXO IgG kit was assessed using a protocol based on the guidelines given in Clinical and Laboratory Standards CLSI document EP17 A In one case 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 Cg is derived from 95 percentile of the standard Gaussian distribution was applied Five low concentration samples of analyte were used determined singly with one batch of r
7. ready for use Reagents 1 2 and 3 are assembled in a single reagents cartridge unit The Calibrator concentrations are expressed in IU mL International Units and calibrated against International Standard 1st IS 01 600 2003 The concentration settings specific for each production lot are recorded on the DATA DISK included in the kit IFU043TGS TA Version 01_EN 10 July 2014 Page 3 of 15 TGS TA TOXO IgG REF YB500032 DATA DISK A Mini DVD 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 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 Cod No TGS00001 IDS iSYS Cuvette Cube Code No IS CC1000 Pack of 960 cuvettes IDS iSYS System Liquid Code No IS CS1000 1 bottle containing 5 liters of ready to use solution TGS TA Wash Solution Code No IS CW1000 1 bottle containing 10 liters of ready to use solution IDS iSYS 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 containi
8. sera at differing levels of concentration at least one negative serum and one positive 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 IFU043TGS TA Version 01_EN 10 July 2014 Page 8 of 15 TGS TA TOXO IgG REF YB500032 CALCULATION AND INTERPRETATION OF THE RESULTS Calculation of the results The concentration of the anti Toxoplasma gondii 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
9. city Cross reactions In order to assess potential cross reactions of the antigen used to sensitise the microparticles a study was conducted using 202 samples all with IgG antibodies positive for Herpes antigens and TORCH complex antigens and IgG antibodies negative for Toxoplasma The samples used were subdivided as follows 40 CMV 40 Rubella 35 EBV VCA G 24 EBV EBNA 1 10 EBV EA D 15 Measles 16 Mumps 12 VZV 9 HSV 1 1 HSV 2 The study did not show any significant cross reaction between the solid phase antigen and 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 TOXO IgG kit since it uses two incubations is not subject to this effect A sample with an extremely high concentration above the measurement range of anti Toxoplasma gondii IgG confirmed the absence of the hook effect up to the concentration of 488 IU mL Relative Sensitivity and Specificity The presence of anti Toxoplasma gondii IgG antibodies was determined using the TGS TA TOXO IgG kit and an immunochemiluminescence assay available on the market in 597 samples 18 samples gave rise to discordant results between the TGS TA assay and the other available method The relative agreement was therefore found to be 97 0 95 Confidence Interval 95 2
10. dentify 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 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 automatically 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 IFU043TGS TA Version 01_EN 10 July 2014 Page 7 of 15 TGS TA TOXO IgG REF YB500032 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 mas
11. eagents in 17 different tests The Limit of Detection of the TGS TA TOXO IgG kit proved to be 0 9 IU mL In the other protocol calculation of the Minimum Detectable Concentration MDC is envisaged 20 replicates of the solution of the 0 IU mL Standard of the Master curve were used The average and the standard deviation SD were formulation on a lot of the kit The RLU value related to the average 2 6 SD 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 2 IU 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 IFU043TGS TA Version 01_EN 10 July 2014 Page 13 of 15 TGS TA TOXO IgG REF YB500032 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 alae Intertenng 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 Analytical Specifi
12. 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 Toxoplasma gondii 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 instructions for use provided The IgG anti Toxoplasma gondii antibody concentration in the TGS TA controls are recorded in the DATA DISK and automatically transferred to the analyser Loading of samples Place the samples in the samples area of the analyser see the analyser user manual on how to i
13. hing of particles measurement of the emitted light IFU043TGS TA Version 01_EN 10 July 2014 Page 2 of 15 TGS TA TOXO IgG REF YB500032 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 purified Toxoplasma gondii antigen 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 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 Human serum with low concentration of anti Toxoplasma gondii 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 concentration of anti Toxoplasma gondii IgG antibodies in Phosphate Buffer containing bovine serum albumin a surfactant an inert blue colouring agent Pro Clin 300 and Gentamicin SO as preservatives All reagents are
14. ignificant source of human infections in some countries Fecal contamination of hands is a significant risk factor Toxoplasmosis is one of the most chronic infections affecting one third of the world s human population The prevalence of Toxoplasma gondii infection varies among different geographical regions The infection is characterized by non specific symptoms with the consequent formation of cysts that may remain in latent form in many organs Primary infection is usually subclinical but in some patients cervical lymphadenopathy or ocular disease can be present There are four groups of individuals in whom the diagnosis of toxoplasmosis is most critical pregnant women who acquire their infection during gestation fetuses and newborns who are congenitally infected immune compromised patients and those with chorioretinitis Although these infections are usually either asymptomatic or associated with self limited symptoms in adults e g fever malaise and lymphadenopathy infections in pregnant women can cause serious health problems in the fetus if the parasites are transmitted i e congenital toxoplasmosis and cause severe IFU043TGS TA Version 01_EN 10 July 2014 Page 1 of 15 TGS TA TOXO IgG REF YB500032 sequelae in the infant including mental retardation blindness and epilepsy The most frequent challenge encountered by physicians the world over is how to determine if a pregnant woman acqu
15. immunological dosages Such samples may give rise to anomalous readings if analysed with the TGS TA TOXO IgG kit IFU043TGS TA Version 01_EN 10 July 2014 Page 9 of 15 TGS TA TOXO IgG REF YB500032 EXPECTED VALUES 89 selected non immune samples have been analysed to check absence of anti Toxoplasma gondii IgG antibodies All samples analysed proved negative with a mean value of 0 0 IU mL and a standard deviation of 0 14 IU mL Using the results obtained the Limit of Blank LoB the highest reading value 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 is equal to 0 3 IU mL with the Reagent lot no 2 DIAGNOSTIC SPECIFICITY AND SENSITIVITY By the use of TGS TA TOXO IgG TGS TA TOXO IgM and TGS TA TOXO IgG Avidity kits it is possible to evaluate the patient immune status related to Toxoplasma infection In the following table are reported the serological profiles of the 3 tests TOXO IgG TOXO IgM e TOXO IgG Avidity and their relative interpretation with respect to the parasite exposure IgG anti IgG anti IgM anti aa Toxoplasma Avidity Interpretation Toxoplasma Toxoplasma 2nd level test Non immune Individual negative negative susceptible to infection ae Suspected acute primary negative positive ea infection ie he Recent primar
16. ington and Klein ed Infectious diseases of the fetus and newborn infant 5 ed W B Saunders Philadelphia Pa 9 Thulliez P Daffos F Forestier f Diagnosis of Toxoplasma infection in pregnant woman and the unborn child current problems Scand J Infect Dis Suppl 1992 84 18 22 10 Montoya JG Remington JS Toxoplasma gondii In Mandel GL Bennett JE Dolin R eds Mandell Douglas and Bennett Principles and Practice of Infectious Diseases 5thg Ed Philadelphia Churchill Livingstone 2000 pp 2858 2888 11 Wilson M McAuley JM Toxoplasma In Murray PR Baron ES Pfaller MA et al eds Manual of Clinical Microbiology 7 Ed Washington DC ASM Press 1999 pp 1374 1382 12 Hedman K Lappalainen M Seppaia Makela O Recent primary toxoplasma infection indicated by a low avidity of specific IgG J Infect Dis 1989 159 4 736 40 13 Montoya JG Remington JS Studies on the serodiagnosis of toxoplasmic lymphadenitis Clin Infect Dis 1995 20 4 781 9 14 Jenun PA Stray Pedersen B Gundersen AG Improved diagnosis of primary Toxoplasma gondii infection in early pregnancy by determination of anti toxoplasma immunoglobulin G avidity J Clin Microbiol 1997 35 8 1972 7 TECHNOGENETICS S r l Via Vanvitelli 4 20129 Milano Italy IFU043TGS TA Version 01_EN 10 July 2014 Page 15 of 15
17. ion took place more than 4 5 months ago 4 PRINCIPLE OF THE METHOD The TGS TA TOXO IgG kit for quantitative determination of specific anti Toxoplasma gondii IgG employs an indirect two step immunological method based on the principle of chemiluminescence 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 During initial incubation the specific antibodies present in the sample in the calibrators or in the controls bind with the solid phase During the second incubation the conjugate reacts with the anti Toxoplasma gondii 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 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 assay protocol addition of samples calibrators controls magnetic particles conjugates and chemiluminescence activation solutions to the reaction cuvette magnetic separation and was
18. ired the acute infection during gestation Women who acquired their infection prior to pregnancy are essentially not at risk for delivering an infected infant unless the woman is immunosuppressed Furthermore methods of diagnosis and their interpretations may differ for each clinical category The diagnosis of Toxoplasma gondii infection may be established by serologic tests amplification of specific nucleic acid sequences PCR histological demonstration of the parasite and or its antigens or by isolation of the organism The use of serologic tests for demonstration of specific antibody to Toxoplasma gondii is the initial and primary method of diagnosis Serological screening relies on IgG and IgM antibody determinations The presence of elevated levels of Toxoplasma specific antibodies indicates infection has occurred at some point but does not distinguish between an infection acquired recently and one acquired in the distant past In acute infection IgG and IgM antibodies generally rise 1 to 2 weeks of infection Detection of Toxoplasma specific IgM antibodies is so used as an aid in determining the time of infection but it is critical to remind that IgM antibodies have been reported to persist for up to 18 months post infection To differentiate between a recently acquired infection and a past infection IgM and IgG positive specimens should be tested for IgG avidity A high avidity index for IgG antibodies is a strong indication that an infect
19. ng 1 liter of ready to use solution IDS iSYS Cartridge Checking System Code No IS 601000 TGS TA ImmunoCleaner Code No YC500012 6 bottles each containing 27 mL 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 da IDS SA 101 103 rue Ernest Solvay 4000 Li ge Belgique and distributed by Technogenetics Srl IFU043TGS TA Version 01_EN 10 July 2014 Page 4 of 15 TGS TA TOXO IgG REF YB500032 Other Recommended Reagents TGS TA TOXO Control Set Code No YB500040 Three 1 5 mL vials of negative human serum and three 1 5 mL vials of human serum positive for anti Toxoplasma gondii antibodies WARNINGS AND PRECAUTIONS The reagents supplied in the TGS TA TOXO IgG 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
20. rom sample collection the method TGS TA TOXO IgG has shown 164 positive samples e 4 negative samples so the sensitivity is 97 6 95 Confidence Interval 93 6 99 2 4 negative samples are of patients with on going seroconversion Among 127 samples from patients diagnosed as infected by Toxoplasma in monitoring and follow up the method TGS TA TOXO IgM has shown 123 positive samples and 4 negative samples so the diagnostic sensitivity was 96 9 95 Confidence Interval 91 6 99 0 By using TGS TA TOXO IgG Avidity kit 100 samples from patients diagnosed as infected within last 4 months from sample collection in monitoring and follow up were tested 94 samples showed low avidity 5 moderate avidity and 1 high avidity Among 41 samples from many patients infected more than last 4 months from sample collection all had positive IgG levels showing diagnostic a sensitivity of 100 positive IgM levels in 39 of patients 16 41 and all 41 samples tested by TGS TA TOXO IgG Avidity kit had high Avidity 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 TOXO IgG kit have been assessed using a protocol based on the guidelines given in Clinical and Laboratory Standards CLSI document
21. rozen 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 using the rack provided and leave it to be agitated for at least 40 minutes before use 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 IFU043TGS TA Version 01_EN 10 July 2014 Page 6 of 15 TGS TA TOXO IgG REF YB500032 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
22. 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 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
23. ter curve for the TGS TA TOXO IgG 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 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 200 uL of conjugate are dispensed into the cuvette The reaction cuvette is incubated in the rotor at 37 C for 10 minutes Par oN 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 Toxoplasma gondii IgG present in the sample 8 The readings obtained are interpolated on the calibration curve and transformed into concentrations Samples with concentration higher than the upper limit of the measurable range can be diluted and retested The new value obtained is multiplied to obtain the final result by the dilution factor used QUALITY CONTROL To ensure the validity of the assay control
24. y infection last 4 positive positive Low Avidity months from sample collection Recent previous Primary x r ae infection with IgM presence positive positive High Avidity more than last 4 months from sample collection positive negative High Avidity Past infection Using the TGS TA TOXO IgG TGS TA TOXO IgM TGS TA TOXO IgG Avidity tests the diagnostic specificity and sensitivity was assessed in the following population of selected samples examined with different methods and classified according to the rule of general consensus 139 non immune patients 127 samples from 61 patients diagnosed as infected by Toxoplasma in monitoring and follow up 41 samples from many patients known as infected more than 4 months from sample collection IFU043TGS TA Version 01_EN 10 July 2014 Page 10 of 15 TGS TA TOXO IgG REF YB500032 Following the interpretation given above it was possible to work out the diagnostic specificity and sensitivity relating to each group under evaluation All 139 non immune patients have shown both IgG and IgM values below cut off by using methods TGS TA TOXO IgG and TGS TA TOXO IgM The diagnostic specificity of methods for IgG and IgM related to non immune patients was 100 95 Confidence Interval 95 9 100 0 Among 168 samples 127 patients diagnosed as infected by Toxoplasma in monitoring and follow up and 41 from many patients known as infected more than 4 months f
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