AIA-600II Assay Specifications TgAb Test Code 048

Transcription

1 AIA-600II Assay Specifications TgAb Test Code 048 Screen Item Data Screen 1 Lot ***Enter current cal. lot no. Cal 1 0 IU/mL Cal 2 2 IU/mL (example) Cal 3 4 IU/mL (example) Cal 4 10 IU/mL (example) Cal 5 20 IU/mL (example) Cal 6 40 IU/mL (example) Screen 2 Name TgAb Unit IU/mL Smpl 50 Dil 100 2Reag 100 Code 48 Assay Range Low 0.24 Assay Range High 40 Reference Range Low Reference Range High DP (No. of decimal points) 2 Screen 3 Screen 4 Code 48 No. (Calibrators) 6 Mul. (Replicates) 3 Equ 3 CV (Calibration curve stability) 90 STAT (Analyte status) 0 PRCL (Assay Protocol) 3 Dilution Factors: SP1 (Specimen 1) 51 SP2 (Specimen 2) 1 CAL 1 CTRL 1 CODE (SDS code) 847 PR (Dilution mode) 3 Pretreatment: SMPL (Pretreatment sample volume) 0 VOL1 (Pretreatment 1 volume) 0 VOL2 (Pretreatment 2 volume) 0 CODE (Pretreatment code) 0 ANTI-THYROGLOBULIN ANTIBODIES REV AIA-600II ASSAY SPECIFICATIONS

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3 AIA-900 Assay Specifications TgAb Test Code 048 No. Item Data 1 Code 48 2 ACT 0 3 Analyte TgAb 4 Lot ***Enter current cal. lot no. 5 CAL 1 0 IU/mL 6 CAL 2 2 IU/mL (example) 7 CAL 3 4 IU/mL (example) 8 CAL 4 10 IU/mL (example) 9 CAL 5 20 IU/mL (example) 10 CAL 6 40 IU/mL (example) 11 Cal lot L 12 Cal lot R 13 Unit IU/mL 14 Decimal 2 15 Assay Low Assay High Reference Low 0 18 Reference High 0 19 Reschedule Low Reschedule High Factor A 1 22 Factor B 0 23 Sample Volume Diluent Volume Step reagent dispensing volume Calibration code CAL. No CAL. MUL CAL. EQU CAL. CV DIL. SP DIL. SP DIL. CAL. (Calculation of dil ratio of 1 conc.) 34 DIL. CNTL DIL. DO DIL. AH DIL. CALC DIL. CODE DIL. NAME TPOAb 40 DIL. PRTY 3 41 PRE. SPVOL (Vol. of pretreated 0 sample) 42 PRE. 1VOL (Vol. of pretreatment sol-1) 0 43 PRE. 2 VOL (Vol. of pretreatment sol-2) 0 44 PRE. CODE (pretreated sol. code) 0 45 PRE. NAME (pretreated sol. name) 0 46 Protocol 4 47 SYS. F_A 1 48 SYS. F_B 0 49 V. CONC G. ORIGIN 0 ANTI-THYROGLOBULIN ANTIBODIES REV AIA-900 ASSAY SPECIFICATIONS

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5 AIA-1800 Assay Specifications TgAb Test Code 048 No. Item Data 1 Unit IU/mL 2 Decimal places 2 3 Reference low Reference high Reschedule low 12 6 Reschedule high Assay range low Assay range high 40 9 Specimen diluent code Specimen diluent name TPOAb/TgAb 11 Dilution factor for Sp Dilution factor for Sp Dilution factor for Control 1 14 Default multiplier for DO Default multiplier for >H 5 16 Factor A e Factor B e Calibration code Calibrator replicates 3 20 Calibrator lot ***Enter current cal. lot no. 21 Calibrator Concentration 22 Cal IU/mL 23 Cal IU/mL (example) 24 Cal IU/mL (example) 25 Cal IU/mL (example) 26 Cal IU/mL (example) 27 Cal IU/mL (example) Dilution factor for calibrator 1 35 Calibration period Incubation time Assay protocol 4 38 Specimen volume Diluent volume Conjugate volume Diluted conjugate volume 0 42 Pretreatment 0 43 Pretreatment specimen volume 0 44 Pretreatment Reagent code 0 45 Pretreatment Reagent name 0 46 Pretreatment Reagent 1 volume 0 47 Pretreatment Reagent 2 volume 0 48 System Factor 1 49 Calibration Code Check Virtual Concentration Graph Origin Calculation with Dilution Factor Yes ANTI-THYROGLOBULIN ANTIBODIES REV AIA-1800 ASSAY SPECIFICATIONS

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7 AIA-2000 Assay Specifications TgAb Test Code 048 No. Item Data 1 Unit IU/mL 2 Decimal places 2 3 Reference low Reference high Reschedule low 12 6 Reschedule high Assay range low Assay range high 40 9 Specimen diluent code Specimen diluent name TPOAb/TgAb 11 Dilution factor for Sp Dilution factor for Sp Dilution factor for Control 1 14 Default multiplier for DO Default multiplier for >H 5 16 Factor A e Factor B e Calibration code Calibrator replicates 3 20 Calibrator lot ***Enter current cal. lot no. 21 Calibrator Concentration 22 Cal IU/mL 23 Cal IU/mL (example) 24 Cal IU/mL (example) 25 Cal IU/mL (example) 26 Cal IU/mL (example) 27 Cal IU/mL (example) Dilution factor for calibrator 1 35 Calibration period Incubation time Assay protocol 4 38 Specimen volume Diluent volume Conjugate volume Diluted conjugate volume 0 42 Pretreatment 0 43 Pretreatment specimen volume 0 44 Pretreatment Reagent code 0 45 Pretreatment Reagent name 0 46 Pretreatment Reagent 1 volume 0 47 Pretreatment Reagent 2 volume 0 48 System Factor 1 49 Calibration Code Check Virtual Concentration Graph Origin Calculation with Dilution Factor Yes ANTI-THYROGLOBULIN ANTIBODIES REV AIA-2000 ASSAY SPECIFICATIONS

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9 ANTI-THYROGLOBULIN ANTIBODIES AIA-PACK TgAb Name and Intended Use AIA-PACK TgAb is designed for IN VITRO DIAGNOSTIC USE ONLY for the quantitative measurement of anti-thyroglobulin antibodies (TgAb) in human serum or heparinized plasma on specific Tosoh AIA System analyzers. Summary and Explanation of Test Primary thyroid disease typically develops as the result of an autoimmune process in which the body produces antibodies against thyroid-specific hormones and proteins. This phenomenon is associated with the autoimmune destructive process - the end result of which is thyroid dysfunction 1. The main component of the colloid contained in the thyroid follicles is the protein thyroglobulin (Tg). Tg, with a molecular weight of 660 kda, is actually synthesized in the thyroid cells and acts as the principle storage protein for T3, T4, and their precursors. Measurement of antibodies to thyroglobulin (TgAb) has proven clinically useful in diagnosing autoimmune thyroid diseases such as Graves disease, Hashimoto s thyroiditis, myxedema, and asymptomatic thyroiditis, particularly in conjunction with antibodies against thyroid peroxidase (TPOAb). 2,3 However, levels of these two antibodies may vary independently. The presence of TgAb during pregnancy has been shown to correlate with post-partum thyroid dysfunction. 4,5 Principle of the Assay The AIA-PACK TgAb is a two-step immunoenzymometric assay which is performed entirely in the AIA-PACK test cup. Anti-thyroglobulin antibody present in the test sample is bound to thyroglobulin antigen immobilized on magnetic solid phase beads. The magnetic beads are washed to remove unbound antibody complexes and are then incubated with enzyme-labeled monoclonal anti-human IgG antibody. The beads are washed again to remove unbound enzymelabeled monoclonal antibody and are then incubated with a fluorogenic substrate, 4- methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the TgAb concentration in the test sample. A standard curve is constructed, and unknown sample TgAb concentrations are calculated using this curve. Material Provided (AIA-PACK TgAb, Cat. No ) 5 trays x 20 test cups (AIA-PACK TgAb Cup) Plastic test cups containing lyophilized magnetic beads coated with Tg antigen with 0.1% sodium azide as a preservative. 5 vials x 5 ml (AIA-PACK TgAb Conjugate) Vials containing liquid monoclonal anti-human IgG antibody conjugated to bovine alkaline phosphatase with 0.1% sodium azide as a preservative. ANTI-THYROGLOBULIN ANTIBODIES REV

10 Materials Required But Not Provided The following materials are not provided but are required to perform anti-thyroglobulin antibody analysis using the AIA-PACK TgAb (Cat. No ) on specific Tosoh AIA Systems. They are available separately from Tosoh. Materials Cat. No. AIA-SYSTEMS: AIA-600II AIA-600II BCR AIA AIA-900 9tray Sorter AIA tray Sorter AIA-1800 ST AIA-1800 LA AIA-2000 ST AIA-2000 LA AIA-PACK: AIA-PACK Substrate Set II AIA-PACK Substrate/Reconstituent AIA-PACK TgAb Calibrator Set Calibrator #1 0 IU/mL Calibrator #2 2 IU/mL (approx.) Calibrator #3 4 IU/mL (approx.) Calibrator #4 10 IU/mL (approx.) Calibrator #5 20 IU/mL (approx.) Calibrator #6 40 IU/mL (approx.) AIA-PACK TPOAb/TgAb Sample Diluting Solution AIA-PACK TgAb Control Set AIA-PACK TgAb Additional Conjugate F AIA-PACK Wash Concentrate Set AIA-PACK Diluent Concentrate Set AIA-PACK Detector Standardization Test Cups AIA-PACK Sample Treatment Cups Sample Cups ADDITIONAL REQUIREMENTS: PIPETTE TIPS (1000/PKG) TIP RACK (EMPTY) PRELOADED PIPETTE TIPS (96 TIPS X 50 RACKS) PRELOADED PIPETTE TIPS (96 TIPS X 5 RACKS) Only materials obtained from Tosoh should be used. Materials obtained elsewhere should not be substituted since assay performance is characterized based strictly on Tosoh materials. 2 REV AIA-PACK TgAb

11 Warnings and Precautions The AIA-PACK TgAb is intended for in vitro diagnostic use only. TgAb Conjugate of different lots should not be mixed. The AIA-PACK TgAb assay should be performed using the conjugate with test cups of the same lot and not with different lots of test cups. The AIA-PACK TgAb contains sodium azide, which may react with lead or copper plumbing to form potentially explosive metal azides. When disposing of such reagents, always flush with large volumes of water to prevent azide build-up. Human biological material used in the preparation of this product has been tested by FDA cleared methods and found negative for the presence of HBsAg and antibody to HIV-1 and HCV. Because no test method can offer complete assurance that products derived from human blood will not transmit infectious agents, it is recommended that this product be handled with the same precautions used for patient specimens. Do not use beyond the expiration date. Storage and Stability All unopened materials are stable until the expiration date on the label when stored at the specified temperature. Materials Cat. No. Refrigerator Temperature (2-8 C): AIA-PACK TgAb (Cup and Conjugate) AIA-PACK TgAb Calibrator Set AIA-PACK TgAb Control Set AIA-PACK TgAb Additional Conjugate F AIA-PACK TPOAb/TgAb Sample Diluting Solution AIA-PACK Substrate Set II AIA-PACK Wash Concentrate AIA-PACK Diluent Concentrate Room Temperature (18-25 C): AIA-PACK Detector Standardization Test Cups AIA-PACK Sample Treatment Cups AIA-PACK test cups may be stored at room temperature (18-25 C) for up to 24 hours. Once opened, TgAb Conjugate is stable for 7 days in the refrigerator (2-8 C). It is strongly recommended that this conjugate be kept refrigerated when not in use. Calibrators must be kept tightly sealed and refrigerated at (2-8 C). After opening or reconstituting, calibrators should be used within 24 hours. After opening, Sample Diluting Solution is stable for up to 90 days refrigerated at 2-8 C. Reconstituted substrate solution is stable for 3 days at C or 30 days at 2-8 C. Working diluent and wash solutions are stable for 30 days at room temperature (18-25 C). Reagents should not be used if they appear cloudy or discolored. ANTI-THYROGLOBULIN ANTIBODIES REV

12 Specimen Collection and Handling Serum or heparinized plasma is required for the assay. EDTA and citrated plasma SHOULD NOT BE USED. No special patient preparation is necessary. When using serum, a venous blood sample is collected aseptically without additives (Red top tube). Store at C until a clot has formed (usually minutes), then centrifuge to obtain the serum specimen for assay. SST or gel tubes have not been validated. To use heparinized plasma, a venous blood sample is collected aseptically with the designated additive. Centrifuge and separate plasma from the packed cells as soon as possible. Samples may be stored at 2-8 C for up to 24 hours prior to analysis. If the analysis cannot be done within 24 hours, the sample should be stored frozen at -20 C or below for up to 60 days. Repeated freeze-thaw cycles should be avoided. Turbid serum samples or samples containing particulate matter should be centrifuged prior to testing. Prior to assay, slowly bring frozen samples to room temperature (18-25 C) and mix gently. All patient specimens require dilution prior to analysis. Samples are diluted 1:51 with TPOAb/TgAb Sample Diluting Solution. The dilution factor is pre-programmed in the software. The AIA-System will automatically perform these dilutions and calculate the results. TgAb calibrators and control materials do not require dilution prior to assay. The sample required for analysis is 50 µl of serum. Procedure 1) Reagent Preparation 1a) Substrate Solution Bring all reagents to room temperature (18-25 C) before preparing the working reagent. Add the entire contents of the Substrate Reconstituent (100 ml) to the lyophilized Substrate and mix thoroughly to dissolve the solid material. 1b) Wash Solution Add the entire contents of the Wash Concentrate (100 ml) to approximately 2.0 L of CAP Class I or NCCLS (CLSI) Type I Reagent Grade water, mix well, and adjust the final volume to 2.5 L. 1c) Diluent Add the entire contents of the Diluent Concentrate (100 ml) to approximately 4.0 L of CAP Class I or NCCLS (CLSI) Type I Reagent Grade water, mix well, and adjust the final volume to 5.0 L. 1d) TgAb Conjugate The conjugate is provided ready to use. 1e) TPOAb/TgAb Sample Diluting Solution The Sample Diluting Solution (SDS) is provided ready to use. 4 REV AIA-PACK TgAb

13 2) Calibration 2a) Calibration Curve The calibrators for use with the AIA-PACK TgAb are referenced against the NIBSC standard (Code 65/93). The calibration curve for the AIA-PACK TgAb is stable for up to 90 days. Calibration stability is monitored by quality control performance and is dependent on proper reagent handling and AIA System maintenance according to the manufacturer s instructions. Recalibration may be necessary more frequently if controls are out of the established range for this assay or if certain service procedures are performed (e.g. temperature adjustment, sampling mechanism changes, or detector lamp adjustment or change). For further information regarding instrument operation, consult the AIA System Operator s Manual. The sample calibration curve below shows the algorithm used to calculate the results. This is an example only. Actual results will vary depending on the Instrument type and lot number used. ANTI-THYROGLOBULIN ANTIBODIES REV

14 2b) Calibration Procedure i) Refer to the appropriate AIA System Operator s Manual for procedural instructions. ii) Verify that both lots of AIA-PACK TgAb Test Cup and Conjugate are the same. iii) Verify that both the calibrator lot and concentration numbers have been correctly entered into the software. iv) Calibrators for AIA-PACK TgAb are lyophilized with the exception of the zero calibrator. All other levels should be reconstituted with 1 ml of CAP Class I or NCCLS (CLSI) Type I Reagent Grade water. Tosoh recommends that all calibrators be run in triplicate. v) Verify that the TgAb conjugate has been placed on the reagent cassette in the position designated under REAGENT REGISTRATION. For detailed instructions regarding REAGENT REGISTRATION, please refer to the AIA System Operator s Manual. 2c) Calibration Acceptability criteria i) The mean rate for the Calibrator (1) should be < 2.0 nm/sec. ii) Since there is a direct relationship between concentration and rate, the rates should increase as the concentration increases. iii) The replicate values should be within a 10% range. 2d) Calibration Review and Acceptance i) Using the criteria above, review the calibration curve carefully. ii) Edit the calibration if necessary and then accept the calibration. iii) For further information regarding calibration, consult the specific AIA System Operator s Manual. 3) Quality Control 3a) Commercially Available Controls Commercially available controls should be run at least once per day. It is recommended that at least two (2) levels of controls, normal and abnormal, be used. Laboratory policy for this particular assay designates the following: Control Material: Frequency: Lot number of control material, acceptable limits, and corrective action to be taken if controls do not meet laboratory criteria will be found in a separate quality control document maintained by the laboratory. 6 REV AIA-PACK TgAb

15 3b) Quality Control Procedure i) Assay quality control specimens as instructed in the specific Operator s Manual for your analyzer. In addition, refer to the AIA System Operator s Manual for detailed instructions on defining and editing the files. ii) Quality control material to be run with this assay is defined by individual laboratory policy. 4) Specimen Processing 4a) Preparation Following specific instructions in the Operator s Manual for your analyzer. Place samples on the instrument appropriately. 4b) Assay Procedure i) Ensure a sufficient quantity of ST AIA-PACK TgAb test cups for the number of samples to be run. ii) Load patient samples as instructed in the Operator s Manual and proceed with analysis. Note: The AIA-AIA-900, AIA-600II, AIA-1800 and AIA-2000 will require AIA- PACK Sample Treatment Cups if onboard dilutions are utilized. iii) Verify that both the TgAb conjugate and the TPOAb/TgAb Sample Diluting Solution are on the reagent cassette in the positions designated under REAGENT REGISTRATION. For detailed instructions regarding REAGENT REGISTRATION, please refer to the AIA System Operator s Manual. Procedural Notes 1. Lyophilized Substrate must be completely dissolved. 2. Ligand assays performed by the Tosoh AIA Systems require that the laboratory use water designated by the College of American Pathologists as Class I or by NCCLS (CLSI) as Type I. Water should be tested at least once per month and should be free of particulate matter including bacteria. The ph of the water should also be routinely tested. For further information, consult the NCCLS (CLSI) document Preparation and Testing of Reagent Water in the Clinical Laboratory, NCCLS (CLSI) Document C3-A2, Volume 11 No. 13 originally approved as a guideline by NCCLS in August If a specimen anti-thyroglobulin antibody concentration after diluting 51 fold is found to be greater than the linearity limit of the assay, 40 IU/mL (2040 IU/mL after multiplying by dilution factor of 51); the specimen should be diluted further with the AIA-PACK TPOAb/TgAb Sample Diluting Solution and re-assayed according to the Assay Procedure. The recommended further dilution for samples containing greater than 40 IU/mL (2040 IU/mL after multiplying by dilution factor of 51) is 1:10. It is desirable to dilute the sample so that the diluted sample reads between 0.3 IU/mL (12 IU/mL after multiplying by dilution factor of 51) and 40 IU/mL (2040 IU/mL after multiplying by dilution factor of 51). If the autodilution feature is not being used, further dilutions must be made using the original 1:51 dilution. For further information on the dilution of specimens, refer to the AIA System Operator s Manual. ANTI-THYROGLOBULIN ANTIBODIES REV

16 4. The AIA systems can store two different calibration curves for each analyte at one time. However, only one lot of TgAb test cups should be used during the same run since the TgAb conjugate in use should be the same lot number as the test cups. 5. If the assay specifications for this test are not already in the system software, the specifications must be entered under test code 48. Calculation of Results The AIA Systems perform all sample and reagent handling operations automatically. The AIA Systems read the rate of fluorescence produced by the reaction and automatically convert the rate to anti-thyroglobulin antibody concentration in IU/mL. For samples requiring dilution, the AIA AIA-900, AIA-600II, AIA-1800 and AIA-2000 will automatically perform dilutions and calculate results if the dilution factors are entered into the software. Dilution factors may be entered when programming specimens in Assay Monitor. Evaluation of Results Quality Control In order to monitor and evaluate the precision of the analytical performance, it is recommended that commercially available control samples be assayed daily. The minimum recommendations for the frequency of running internal control material are: After calibration, a minimum of two levels of controls are run in order to accept the calibration curve. The two levels of controls are also repeated after calibration when certain service procedures are performed (e.g. temperature adjustment, sampling mechanism changes, maintenance of the wash probe or detector lamp adjustment or change). After daily maintenance, at least two levels of the control should be run in order to verify the overall performance of the Tosoh AIA System Analyzers. If one or more control sample value(s) is out of the acceptable range, it will be necessary to investigate the validity of the calibration curve before reporting patient results. Standard laboratory procedures should be followed in accordance with the regulatory agency under which the laboratory operates. Limitations of the Procedure For diagnostic purposes, the results obtained from this assay should be used in conjunction with other data (e.g., symptoms, results of other tests, clinical impressions, therapy, etc.). Using the AIA-PACK TgAb, the highest concentration of anti-thyroglobulin antibody measurable is approximately 40 IU/mL (2040 IU/mL after multiplying by dilution factor of 51), and the lowest measurable concentration is 0.24 IU/mL (12 IU/mL after multiplying by dilution factor of 51). Although the approximate value of the highest calibrator is 40 IU/mL (2040 IU/mL after multiplying by dilution factor of 51), the exact concentration may be slightly different. The assay specification, Assay Range High, should be defined as the upper limit of the assay range, 40 IU/mL (2040 IU/mL after multiplying by dilution factor of 51). 8 REV AIA-PACK TgAb

17 Although hemolysis has an insignificant effect on the assay, hemolyzed samples may indicate mistreatment of a specimen prior to assay and results should be interpreted with caution. Lipemia has an insignificant effect on the assay except in the case of gross lipemia where spatial interference may occur. Certain medications may interfere with assay performance. All results should be interpreted with respect to the clinical picture of the patient. For a more complete understanding of the limitations of this procedure, please refer to the Specimen Collection and Handling, Warnings and Precautions, Storage and Stability, and Procedural Notes sections in this insert sheet. Expected Values Each laboratory should determine a reference interval which corresponds to the characteristics of the population being tested. As with all diagnostic procedures, clinical results must be interpreted with regard to concomitant medications administered to the patient. 6 Reference Ranges In a group of serum samples from 421 apparently healthy individuals, 97.5% of the values were found to be less than 28.7 IU/mL. Reference interval = <28.7 IU/mL Performance Characteristics The following performance characteristics were determined using the Tosoh AIA-1200 Series Automated Immunoassay Analyzer. The AIA-systems demonstrate equivalent performance. 1) Accuracy 1a) Recovery: Three samples were spiked with three different levels of TgAb and assayed before and after spiking. Initial Value Sample (IU/mL) Sample A Sample B Sample C TgAb Added (IU/mL) Expected Value (IU/mL) Measured Value (IU/mL) Percent Recovery (%) ANTI-THYROGLOBULIN ANTIBODIES REV

18 1b) Dilution: Three samples which had been diluted by 51 fold were serially diluted with AIA-PACK TPOAb/TgAb Sample Diluting Solution and assayed. Sample Sample A2 Sample B2 Sample C2 Dilution Factor None 7.5/10 5.0/10 2.5/10 1.0/10 None 7.5/10 5.0/10 2.5/10 1.0/10 None 7.5/10 5.0/10 2.5/10 1.0/10 Expected Value (IU/mL) Measured Value (IU/mL) Percent Recovery (%) c) Comparative Analysis: 67 serum samples were analyzed with the AIA-PACK TgAb assay and another commercially available anti-thyroglobulin assay. The statistics, presented below, demonstrate good correlation between these two methods. Slope y-intercept 1.57 IU/mL Correlation Coefficient Range of Samples IU/mL Number of Samples (n) 67 2) Precision 2a) Intra-assay precision The intra-assay (within run) precision coefficient of variation was evaluated in three samples by 10 replicate determinations. Sample Sample A Sample B Sample C Number of Replicates Mean (IU/mL) Standard Deviation (IU/mL) Coefficient of Variation (%) REV AIA-PACK TgAb

19 Inter-assay precision The inter-assay (between run) precision coefficient of variation was evaluated at two different concentrations by analyzing samples in 20 separate runs. Sample Sample A Sample B Number of Replicates Mean (IU/mL) Standard Deviation (IU/mL) Coefficient of Variation (%) Sensitivity The minimal detectable concentration (MDC) of human anti-thyroglobulin antibody is estimated to be 0.24 IU/mL which corresponds to 12 IU/mL in original serum sample (dilution factor 51). The MDC is defined as that concentration of human TgAb which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. Interference Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. The results were obtained by adding the interfering substances to diluted samples. Hemoglobin (up to 450 mg/dl), free bilirubin (up to 13 mg/dl) and conjugated bilirubin (up to 19 mg/dl) do not interfere with the assay. Lipemia, as indicated by added triglyceride concentrations (up to 290 mg/dl), does not interfere with the assay. Protein, as indicated by added albumin concentrations (up to 5 g/dl), does not interfere with the assay. Ascorbic acid (up to 20 mg/dl) does not interfere with the assay. ANTI-THYROGLOBULIN ANTIBODIES REV

20 References 1. Clinical Utility of Thyroid Autoantibody Measurement. In: Standards of Laboratory Practice, Laboratory Support of the Diagnosis & Monitoring of Thyroid Disease. L.A. Kaplan and C.T. Sawin eds. National Academy of Clinical Biochemistry Monograph. p.39 (1996). 2. Feldt-Rasmussen, U., Analytical and Clinical Performance Goals for Testing Autoantibodies to Thyroperoxidase, Thyroglobulin, and Thyrotropin Receptor. Clin Chem 42(1): (1996). 3. Salvi, M., et al., Role of Autoantibodies in the Pathogenesis and Association of Endocrine Disorders. Endocrine Rev 9:450 (1988). 4. Stagnaro-Green A., et al., A Prospective Study of Lymphocyte-initiated Immuosuppression in Normal Pregnancy: Evidence of a T-cell Etiology for Postpartum Thyroid Dysfunction. J. Cin Endocrinol. Metab. 74:645 (1992). 5. Tamaki, H., et al., Usefulness of thyroglobulin antibody Detected by Ultrasensitive Immunoassay: A Good Parameter for Immune Surveillance in Healthy Subjects and for Prediction of Post-partum Thyroid Dysfunction. Clin Endocrinol. 37: (1992). 6. Young, D., Effects of Drugs on Clinical Laboratory Tests. 3rd Edition, American Association for Clinical Chemistry Press REV AIA-PACK TgAb

21 AIA-PACK TgAb Calibrator Set Intended Use The AIA-PACK TgAb Calibrator Set is intended for IN VITRO diagnostic use only for the calibration of the AIA-PACK TgAb Assay. Summary and Explanation The AIA-PACK TgAb Calibrator Set contains buffered bovine serum albumin with assigned levels of anti-thyroglobulin antibodies. Calibration should be performed according to the schedule indicated in the AIA System Operator's Manual. The calibrators in this set have been referenced against the NIBSC standard (Code 65/93). Material Provided (Cat. No ) 2 x 1mL Calibrator (1) 0 IU/mL Buffered bovine serum albumin containing no detectable concentration of TgAb (0 IU TgAb/mL), and 0.1% sodium azide as a preservative. 2 x 1mL Calibrator (2) 2 IU/mL (approx.) Calibrator (3) 4 IU/mL (approx.) Calibrator (4) 10 IU/mL (approx.) Calibrator (5) 20 IU/mL (approx.) Calibrator (6) 40 IU/mL (approx.) Buffered bovine serum albumin containing the assigned concentration of TgAb (described on each vial). (Lyophilized) Warnings and Precautions The AIA-PACK TgAb Calibrator Set is for IN VITRO diagnostic use. Calibrator (1) contains sodium azide, which may react with lead or copper plumbing to form potentially explosive metal azides. When disposing of such reagents, always flush with large volumes of water to prevent azide build-up. Although material derived from human blood is not used for TgAb Calibrators, it is recommended that this product be handled with the same precautions used for patient samples. Do not use beyond the expiration date. ANTI-THYROGLOBULIN ANTIBODIES REV

22 Preparation and Storage The CALIBRATOR (1) is provided ready to use. The AIA-PACK TgAb CALIBRATOR SET does not require dilution with SAMPLE DILUTING SOLUTION. Using volumetric pipettes, reconstitute the lyophilized calibrators (2 to 6) accurately to the volume of 1 ml with CAP Class I or NCCLS (CLSI) Type I Reagent Grade water. Allow the lyophilized material to fully dissolve, then mix the calibrators gently but thoroughly prior to performing the calibration. Bring calibrator to room temperature (18-25 C) for use. Always store the Calibrator Set in an upright position at 2-8 C when not in use. Stability When stored unopened and refrigerated at 2-8 C, the AIA-PACK TgAb Calibrator Set is stable until the expiration date on the label. After opening, and reconstituting the calibrators should be used within 24 hours. Procedure Refer to the CALIBRATION PROCEDURE in the AIA-PACK section of this analyte application. For additional procedural instructions regarding calibration, refer to the AIA System Operator s Manual. 1. Verify that both lots of AIA-PACK TgAb Test Cup and Conjugate are the same. 2. Verify that both the calibrator lot and concentration numbers have been correctly entered into the software. 3. Calibrators for AIA-PACK TgAb are lyophilized with the exception of the zero calibrator. All other levels should be reconstituted with 1 ml of CAP Class I or NCCLS (CLSI) Type I Reagent Grade water. 4. Tosoh recommends that all calibrators be run in triplicate. 5. Verify that the TgAb conjugate has been placed on the reagent cassette in the position designated under REAGENT REGISTRATION. For detailed instructions regarding REAGENT REGISTRATION, please refer to the AIA System Operator s Manual. Assignment of Values The AIA-PACK TgAb Calibrator Set contains assigned concentrations of anti-thyroglobulin antibodies. The assigned value is determined on a lot-to-lot basis and is designed to provide an assay calibration range of approximately 0.24 to 40 IU/mL (12 to 2040 IU/mL after multiplying by dilution factor of 51) of TgAb. The calibrators in this set have been referenced against the NIBSC standard (Code 65/93). 2 REV AIA-PACK TgAb CALIBRATOR SET

23 Results Since there is a direct relationship between concentration and rate, the rates should increase as the concentration increases. The replicate values should be within a 10% range. Limitations The AIA-PACK TgAb Calibrator Set is designed solely for use with AIA-PACK assay procedures. References 1. AIA Analyte Application Manual. Tosoh Bioscience, Inc., South San Francisco, CA. 2. AIA-System Operator s Manual. Tosoh Bioscience, Inc., South San Francisco, CA. ANTI-THYROGLOBULIN ANTIBODIES REV

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25 AIA-PACK TPOAb/TgAb Sample Diluting Solution Intended Use The AIA-PACK TPOAb/TgAb Sample Diluting Solution is intended for IN VITRO DIAGNOSTIC USE ONLY to dilute patient samples. Summary and Explanation The AIA-PACK TPOAb/TgAb Sample Diluting Solution contains buffered bovine serum albumin with no detectable concentration of anti-thyroid peroxidase antibodies or anti-thyroglobulin antibodies. This Sample Diluting Solution is to be used only with samples that are being tested for TPOAb and/or TgAb concentrations using the AIA-PACK TPOAb and/or the AIA-PACK TgAb assay. Material Provided (Cat. No ) 4 x 100 ml Sample Diluting Solution Buffered bovine serum albumin containing no detectable concentration of antithyroid peroxidase antibodies or anti-thyroglobulin antibodies (0 IU TPOAb/TgAb/mL ), and 0.1% sodium azide as a preservative. Warnings and Precautions The AIA-PACK TPOAb/TgAb Sample Diluting Solution is for in vitro diagnostic use. These materials contain sodium azide, which may react with lead or copper plumbing to form potentially explosive metal azides. When disposing of such reagents, always flush with large volumes of water to prevent azide build-up. Although material derived from human blood is not used for this product, it is recommended that this product be handled with the same precautions as used for patient samples. Do not use beyond the expiration date. Preparation and Storage The AIA-PACK TPOAb/TgAb Sample Diluting Solution is provided ready to use. Always store the Sample Diluting Solution in an upright position at 2-8 C when not in use. ANTI-THYROGLOBULIN ANTIBODIES REV

26 Stability When stored unopened and refrigerated at 2-8 C, the AIA-PACK TPOAb/TgAb Sample Diluting Solution is stable until the expiration date on the label. After opening, the Sample Diluting Solution is stable for up to 90 days when refrigerated at 2-8 C. Procedure Refer to the AIA System Operator s Manual for additional procedural instructions regarding sample dilution. 1. If a specimen TPOAb and/or TgAb concentration after diluting 51 fold is found to contain greater than the linearity limit of either assay, the specimen should be diluted with the Sample Diluting Solution and assayed according to the Procedure in the AIA-PACK section of the analyte application. 2. The AIA-600 II, AIA AIA-900, AIA-1800 and AIA-2000 will perform dilutions automatically if the dilution factors are entered into the software prior to assaying the diluted sample. 3. The recommended dilution for specimen containing greater than 40 IU/mL (2040 IU/mL after multiplying with the dilution factor of 51) is 1:10 or 1:100. However, it is desirable to dilute the samples so that the diluted sample reads between IU (8 to 1020 IU/mL after multiplying by dilution factor of 51) TPOAb/mL or IU (12 to 2040 IU/mL after multiplying by dilution factor of 51)TgAb/mL Results When an auto-dilution is performed, the AIA instrument will calculate the final result. Limitations The AIA-PACK TPOAb/TgAb Sample Diluting Solution is designed solely for use with AIA-PACK assay procedures. References 1. AIA Analyte Application Manual. Tosoh Bioscience, Inc., South San Francisco, CA. 2. AIA-System Operator s Manual. Tosoh Bioscience, Inc., South San Francisco, CA. 2 REV AIA-PACK TPOAb/TgAb SDS

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29 AIA-PACK TgAb Control Set Intended Use The AIA-PACK TgAb Control Set is intended for IN VITRO diagnostic use only for performing quality control procedures with the AIA-PACK TgAb Assay. Summary and Explanation The AIA-PACK TgAb Control Set contains buffered bovine serum albumin with assigned levels of anti-thyroglobulin antibodies. TgAb controls should be run once each day on which TgAb assays are scheduled. Material Provided (Cat. No ) 6 x 1 ml Control Level 1 6 x 1 ml Control Level 2 Buffered bovine serum albumin containing approximately 4 IU/mL TgAb (Lyophilized). See vial label for the assigned concentration range. Buffered bovine serum albumin containing approximately 20 IU/mL TgAb (Lyophilized). See vial label for the assigned concentration range. Warnings and Precautions The AIA-PACK TgAb Control Set is for in vitro diagnostic use. Although material derived from human blood is not used for this product, it is recommended that this product be handled with the same precautions as used for patient samples. Do not use beyond the expiration date. ANTI-THYROGLOBULIN ANTIBODIES REV

30 Preparation and Storage Using volumetric pipettes, reconstitute the lyophilized controls accurately to the volume of 1 ml with CAP Class I or NCCLS (CLSI) Type 1 Reagent Grade water. Allow the lyophilized material to fully dissolve. Bring control to C for use. Always store the control set in an upright position at 2 8 C when not in use. The AIA-PACK TgAb CONTROL SET does not require dilution with SAMPLE DILUTING SOLUTION. Stability When stored unopened and refrigerated at 2-8 C, the AIA-PACK TgAb Control Set is stable until the expiration date on the label. After opening, the controls should be used within 7 days. Procedure Refer to the ASSAY PROCEDURE in the AIA-PACK section of this analyte application. For additional procedural instructions regarding quality control, refer to the AIA System Operator s Manual. 1. Enter control Ids into the AIA-System. 2. Enter the assay request (refer to the specific Operator s Manual for details) 3. Add the appropriate amounts of each control material to sample cups (refer to the specific Operator s Manual for details). Place the sample cups and test cups in the proper positions on the instrument. 4. Verify that the TgAb conjugate is on the reagent cassette in the position designated in REAGENT REGISTRATION. Assignment of Values The AIA-PACK TgAb Control Set contains assigned concentrations of anti-thyroglobulin antibodies. The assigned concentration range is determined on a lot-to-lot basis and is designed to provide target control levels of approximately 4 and 20 IU/mL (204 and 1020 IU/mL after multiplying by dilution factor of 51) of TgAb. Since the assay values are dependent upon assay procedures as well as several other factors, each laboratory should establish its own range for the assay procedure being monitored. Limitations The AIA-PACK TgAb Control Set is designed solely for use with AIA-PACK assay procedures. 2 REV AIA-PACK TgAb CONTROLS

31 References 1. AIA Analyte Application Manual. Tosoh Bioscience, Inc., South San Francisco, CA. 2. AIA-System Operator s Manual. Tosoh Bioscience, Inc., South San Francisco, CA. ANTI-THYROGLOBULIN ANTIBODIES REV