Part No. J04498 Cat No Performance Verifiers: Training Module

Transcription

1 Cat No : Training Module

2 Export authorized under general license GTDA (General Technical Data Available) IMPORTANT The information contained herein is based on the experience and knowledge relating to the subject matter gained by Ortho-Clinical Diagnostics, Inc. prior to publication. No patent license is granted by the information. Ortho-Clinical Diagnostics, Inc. reserves the right to change this information without notice, and makes no warranty, express or implied, with respect to the information. The company shall not be liable for any loss or damage, including consequential or special damages resulting from the use of this information, even if loss or damage is caused by its negligence or other fault. VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., All rights reserved.

3 Revision History Revision Date Description Added all MicroTip Drugs of Abuse Testing (DAT) Assays Added all MicroTip and MicroSlide Assays and Performance Verifiers that have become available since last release. 8/99 Company name changed to Ortho-Clinical Diagnostics, Inc., a Johnson & Johnson Company VITROS Chemistry Systems iii

4 List of Revised Pages Each page in your guide should be at the date listed below: Revision Date Chapter Page All Chapters All All chapters All 8/99 All chapters All iv VITROS Chemistry Systems

5 About This Training Manual Purpose The purpose of this module is to provide information on how VITROS can be used to monitor performance of VITROS Chemistry Systems. For setting up Quality Control files on your VITROS Chemistry System, refer to the Quality Control section of the VITROS 350/250, 250/250AT, 950/950AT Operator s Guide or the V-DOCs for the 5,1 FS Chemistry System. Audience This module is intended for use by customers of Ortho-Clinical Diagnostics using VITROS 350/250/250AT, 950/950AT and 5,1 FS Chemistry Systems. How This Module Is Organized This module describes, how the range of means and within-lab standard deviations are established, and how Performance Verifiers can be used with VITROS Chemistry Systems. Additional Training Modules Statistical Tools of Quality Control Part No. 7B6006 CAT No Additional training modules are available on orthoclinical.com. For ordering information, please contact your Ortho-Clinical Diagnostics Laboratory Specialist VITROS Chemistry Systems v

6 About This Training Manual vi VITROS Chemistry Systems

7 Contents Purpose v Audience v How This Module Is Organized v Additional Training Modules v Introduction Objectives Definitions What are? How the Range of Means (ROM) is Established How the Range of Means (ROM) is Used How the Within-lab Standard Deviation (Within-lab SD) is Established How the Within-lab Standard Deviation is Used Using Verifying a New Calibration Establishing Baseline Means for VITROS Chemistry Systems vii

8 Contents Monitoring System Performance on a Periodic (Monthly) Basis Verifying a Previous Calibration When System Performance May Have Changed Verifying a Previous Calibration When No Valid Baseline Mean for Has Been Established Appendix 1: Performance Verifier Assay Sheets Appendix 2: Tables Appendix 3: Quality Control Worksheets Exercises Answer Key viii VITROS Chemistry Systems

9 Introduction The purpose of this module is to provide you with recommendations for using to monitor the performance of the VITROS Chemistry System in your laboratory. These recommendations do not, however, exclude alternative approaches to monitoring your system s performance. Prerequisites for this module are: Ability to calculate a mean and standard deviation. Ability to define and apply the 1 2S, 2 2S, and 1 3S quality control multirules. If you have not fulfilled these prerequisites, you must complete the following module: Statistical Tools of Quality Control Module VITROS Chemistry Systems 1

10 Introduction Objectives Upon completion of this module, you will be able to: Explain what are and how to use them. Describe how to establish a mean for each level of Performance Verifier. Explain the information found on the assay sheets which accompany the. Understand Range of Means and Within-lab standard deviation. Perform daily quality control using. Perform calibration verification on a new slide/reagent lot using. Use to monitor your system s performance periodically. Verify pre-existing calibration when there are no valid baseline means available for. This module will take approximately two hours to complete. You will need the following items: Pencil Calculator This module contains practice exercises to help you apply the concepts learned. The answer key for the practice exercises is provided. 2 VITROS Chemistry Systems

11 Definitions Generation (Gen) is a term used to describe those slide lots that can use the same Supplementary Assigned Values (SAVs) for calibration. Slide lots from the same generation have the same Range of Means on the assay sheet. The Gen number for each lot of slides is found on the packaging as well as on the side and top labels of individual slide cartridges. Performance Verifier Range of Means (ROM) is a term used to describe the lowest and highest mean values your VITROS Chemistry System should produce for your slide lot. These ranges are specific for a particular Gen of slides. These ranges define the acceptable mean values for any generation of slides. On the assay sheet for, each Gen of a particular analyte is listed with its corresponding ROM. A Baseline SD is the value the VITROS Chemistry System QC software uses in combination with the mean to calculate the 1 2s, 2 2s and 1 3s control rule limits that should be used for your daily quality control decisions. The Baseline SD that is used to calculate control limits in the VITROS Chemistry System software may be a fixed criteria, such as the within-lab SD from the Performance Verifier assay sheets. This is a calculated SD determined from the laboratory s experience with the VITROS Chemistry Systems, or your laboratory s medically useful limits. The Within-lab SDs on the assay sheets were established using monthly within-lab SD for VITROS Chemistry System users participating in a commercial quality control service. These values are VITROS Chemistry Systems 3

12 Definitions representative of the performance of properly operating VITROS Chemistry Systems. These values are representative of the performance of properly operating VITROS Chemistry Systems in multiple laboratories using multiple Performance Verifier lots. Additionally, each SD was compared to USA and European proficiency testing goals and adjusted accordingly. The published within-lab SD includes the variability associated with performing replicate measurements within a day and measurements from one day to the next. The day-to-day variability includes the small variation introduced by different slide cartridges, different vials of control material, multiple calibration events, environmental influences, and preventive maintenance events. These values do not include the variability due to using different slide lot numbers. A within-lab SD can be calculated from your daily quality control results and compared to the Performance Verifier assay sheets within-lab SD. A calculated laboratory SD larger than the Performance Verifier assay sheets within-lab SD indicates that system troubleshooting may be necessary. If the calculated SD is much smaller than the Performance Verifier assay sheets within-lab SD, you may not have included all of the expected sources of variability or valid QC results may have been excluded from the calculation. If you use this calculated SD as your baseline SD, valid data points may be rejected and troubleshooting may be performed more frequently than needed. Calibration Verification is a means of assessing whether a calibration is acceptable by using results from each level of. At a minimum, these results should be within the ROM for that control. A Preliminary Baseline Mean is the average of two values run sequentially (two replicates from one cup) immediately after the system is calibrated. This should be done using a fresh vial of PV. This mean is used to verify this calibration. This value should be used as the baseline mean in the VITROS Chemistry System QC software until a more robust estimate of the mean is available (refer to your analyzer Operator s Guide or VDocs for the 5,1 FS). If you are not using PVs as your daily QC materials, then record the preliminary baseline mean as a Calibration Verification Mean for use in troubleshooting and verification of a pre-existing calibration. A Baseline Mean is the average of QC results observed over days, and this mean should replace the preliminary baseline mean in VITROS Chemistry System software. It is a better estimate of the true mean than the 2-point preliminary baseline mean (refer to your analyzer Operator s Guide or VDocs for the 5,1 FS). 4 VITROS Chemistry Systems

13 Definitions A Current Mean is the mean value you should use to periodically evaluate the VITROS Chemistry System after you have established a baseline mean and when there is no evidence that anything has happened to alter your system s performance. The Current Mean will typically be your calculated monthly mean (refer to your analyzer Operator s Guide or VDocs for the 5,1 FS). A Current SD is the standard deviation calculated along with your monthly Current Mean (refer to your analyzer Operator s Guide or VDocs for the 5,1 FS). A is the absolute difference between any two means. If the is greater than the values in the Tables in Appendix 2, then the two means are different. Depending on your laboratory s policy and the magnitude of the difference, troubleshooting may be necessary if this is the case. The values in the tables for n = 2 should also be used as guidelines for acceptable replication of in any evaluation. The delta values in the tables were calculated using standard statistical techniques for the comparison of two means. We used the Performance Verifier assay sheet SD as the basis for this comparison. For small sample sizes (n=2), the calculated deltas assure that the probability of concluding that the means are not different when they truly are, is small. For sample sizes of 20 or more, the calculated deltas assure that the likelihood of saying that the means are different when they are truly not, is small. These choices give laboratory personnel a good balance between the risk of making a certain type of error and the effect that increasing sample size has on the quality of the estimate of the difference between the two means VITROS Chemistry Systems 5

14 Definitions 6 VITROS Chemistry Systems

15 What are? (PVs) are assayed controls designed for use as quality control tools. PVs provide you with a way to assess the calibration of your analyzer, monitor the daily performance of the system, and assist you in troubleshooting out-of-control conditions indicated by your daily quality control results. The following are currently available: AAT/HPT Performance Verifier Level I, II and III ApoA1 Performance Verifier Level I ApoB Performance Verifier Level I ASO/RF Performance Verifier Level I and II CRP Performance Verifier Level I and II DAT Performance Verifier Level I, II, III, IV and V dtibc Performance Verifier Level I and II HCY Performance Verifier Level I, II and III hscrp Performance Verifier Level I, II and III Isoenzyme Performance Verifier Level I and II Liquid Performance Verifier Level I and II VITROS Chemistry Systems 7

16 What are? malb Performance Verifier Level I and II PALB Performance Verifier Level I and II Performance Verifier Level I and II Protein Performance Verifier Level I, II and III TDM Performance Verifier Level I, II and III UPRO Performance Verifier Level I and II %A1c Performance Verifier Level I and II The assay sheet which accompanies these fluids includes the following information for each analyte: A generation (Gen-) specific range of means (ROM). A recommended within-lab standard deviation (SD). Appendix 1 shows an example of an assay sheet. How the Range of Means (ROM) is Established How the Range of Means (ROM) is Used The width of the ROM (highest mean value minus the lowest mean value) is a fixed parameter. The ROM reflects the performance of properly operating VITROS Chemistry Systems as monitored with Performance Verifiers. The ROM width for each analyte was compared to USA and European proficiency testing goals and adjusted as appropriate. The numerical values that define the ROM for each slide generation are established by collecting data from internal testing performed on multiple analyzers and among different slide lot numbers within the slide generation. When evaluating the performance of a VITROS Chemistry System using, the mean based on two or more replicate measurements of these fluids must be within the ROM to be acceptable. Since are manufactured fluids, they do not have the same physical and chemical characteristics or matrix as fresh patient specimens. These differences may cause the results on different slide lots to vary. The term matrix effects is commonly used to describe this phenomenon. Matrix effects result in wider ROM than would otherwise be observed. 8 VITROS Chemistry Systems

17 What are? The mean of two or more measurements can fall anywhere in the ROM (not necessarily in the center) for that particular slide Gen. For calibration verification, the mean of 2 or more replicate measurements of (called a Preliminary Baseline Mean) should fall within the ROM listed on the assay sheet. When using for routinely monitoring a properly operating system, all calculated means should be within the ROM. Each estimate of the mean should be statistically the same as all other estimates of the mean. The assessment of any two means will be discussed later on in this module. It is not expected that all individual daily QC values will fall within the ROM even if the system is showing acceptable performance. However, the mean of a distribution of daily QC values for properly operating systems should always fall within the ROM. How the Within-lab Standard Deviation (Within-lab SD) is Established The published within-lab SD on the Performance Verifier assay sheets was established using monthly within-lab SD for VITROS Chemistry System users participating in a commercial quality control service. These values are representative of the performance of properly operating VITROS Chemistry Systems in multiple laboratories using multiple Performance Verifier lots. Additionally, each SD was compared to USA and European proficiency testing goals and adjusted as appropriate. The published within-lab SD on the Performance Verifier assay sheets normally does not change from one lot of to another. However, it may be adjusted if the analyte concentration changes significantly. This should not occur very often. How the Within-lab Standard Deviation is Used The baseline SD that is used to calculate control limits in the VITROS Chemistry System software may be a fixed criteria, such as the within-lab SD from the Performance Verifier assay sheets, a calculated SD determined from the laboratory s experience with the VITROS Chemistry Systems, or your laboratory s medically useful limits. We recommend that you use the within-lab SD from the PV assay sheets for your baseline SD and your daily quality control decisions VITROS Chemistry Systems 9

18 What are? Some laboratories also calculate the actual SD for the VITROS Chemistry System. A calculated laboratory SD larger than the PV assay sheet within-lab SD indicates that system troubleshooting may be necessary. If the calculated SD is much smaller than the PV assay sheet withinlab SD, you may not have included all of the expected sources of variability or valid QC results may have been excluded from the calculation. If you use this calculated SD as your baseline, you may be rejecting valid data points and be troubleshooting more frequently than needed. 10 VITROS Chemistry Systems

19 Using (PVs) can be used: To verify calibration. As daily quality control materials. As tools for troubleshooting. The flowcharts described below illustrate how to use to address circumstances. Flowchart 1: Verifying a new calibration. Flowchart 2: Establishing Baseline Means for PVs (Using PVs as daily QC materials). Flowchart 3: Monitoring System performance on a periodic (monthly) basis. Flowchart 4: Verifying a previous calibration when system performance may have changed. Flowchart 5: Verifying a previous calibration when no valid Baseline Means for have been established VITROS Chemistry Systems 11

20 Using Verifying a New Calibration Those laboratories not using PVs as daily QC materials do not need to use flowchart 2. However, it is assumed that these laboratories will have PVs on hand to assist in troubleshooting and calibration verification. IMPORTANT: To verify a previous calibration when using Performance Verifiers as daily QC, you should always being with Flowchart 3. Some abbreviated language has been used to keep the flowcharts short and simple. Refer to the Definitions section of this module to find explanations for the some terms you will find in the flowcharts that follow. Whenever the means of two or more measurements of Performance Verifiers, made after a new calibration, are within the ROM, these results reflect the distribution of acceptable calibrations for VITROS Chemistry Systems. The mean for a given combination of analyzer, slide lot, and Performance Verifier lot is not expected to fall in the middle of the ROM for that particular generation of slides. For verifying calibration immediately following a new calibration, follow these steps (see Flowchart 1): 1. After calibration, measure in duplicate the PV fluid (from the same cup) from freshly reconstituted vials of PV Level I and PV Level II (also PV Level III, if applicable). 2. Calculate the mean for two replicates of each PV fluid. These are the preliminary baseline means. 3. If the preliminary baseline mean values of all fall within the range of means, calibration is verified. Note: In addition, you may prefer to evaluate the performance of the newly calibrated slide lot by comparing it with a previous slide lot or another method. Compare the results of 5 10 patient samples run on the current and new slide lots with your laboratory s analyte-specific evaluation criteria. 4. Input these preliminary baseline means in the analyzer software or record these mean values in your QC system. Appendix 3 contains a Calibration Verification worksheet that can be used to record these values and document your other QC records. 12 VITROS Chemistry Systems

21 Using Calibrate Slide/Fluid Lot. Run fresh vials of PVs in duplicate. Possible reasons to calibrate: New slide lot New lot of immuno-wash or electrolyte reference fluids Extensive service performed Result of troubleshooting Required by regulations For each PV calculate the mean of these 2 values. Are these means within the published Range of Means? Yes Calibration is verified 2. No Investigate (see Calibration Protocol Checklist below) Calibration Protocol Checklist: Are materials fresh (slide, calibrators, fluids, and controls)? Confirm insert sheet directions for reconstitution were followed. Are calibrator responses similar to previous responses? Call Technical Support Do you use PVs as your daily QC materials? Yes Input these means as your Preliminary Baseline Means in the VITROS System software or record in your QC system 3. No Record these means in your QC system 3 as your Calibration Verification means. 1 Refer to Definitions. Use the table that corresponds to the appropriate Performance Verifier product. 2 Some accreditation agencies and state regulations may require additional protocols. Comply as required. 3 See Appendix 3. Flowchart 1. Verifying a New Calibration Using Performance Verifiers (PVs) VITROS Chemistry Systems 13

22 Using Establishing Baseline Means for If you intend to use as your daily quality control materials, you need to establish a baseline mean. This is necessary because the baseline mean established using additional data points is a better estimate of the true mean than the 2-point preliminary baseline mean. The more data collected to calculate the baseline mean, the more confidence you will have that your calculated mean approaches the true mean. To calculate a baseline mean you must run for a minimum of 20 days. This length of time will help ensure that multiple vials of and multiple slide cartridges are included in this estimate. During this period, you must change the slide cartridges and control fluid vials at least twice. We recommend that you establish the baseline mean for a new lot of while still using your current lot of Performance Verifiers to make daily QC decisions. If you need to use a new lot of for daily QC decisions while establishing the baseline mean, use the preliminary baseline mean established during calibration verification and the Withinlab SD as the criteria for making your QC decisions. Also use the 13S rule (only react to the F3 QC flags from the VITROS Chemistry System software) during this period. The baseline mean should not differ from the preliminary baseline mean by more than the for n = 2 from Appendix 2. Appendix 3 contains a Baseline Mean Evaluation worksheet to record these values. See Flowchart 2 for an outline of this process. 14 VITROS Chemistry Systems

23 Using If necessary, use your Preliminary Baseline Means and the PV Within-lab SDs for making QC decisions. 1 Apply 1 3S rule only until the Baseline Means are established. 2 Collect quality control data on each PV over a day period. For each PV calculate the Baseline Mean 3. Are these means within the published Range of Means? No Yes Investigation required: Contact Technical Support Are the differences between Preliminary Baseline Means and Baseline Means the delta 3 (for n=2) in Appendix 2? No Yes Use these Baseline Means and your laboratory specific QC procedures to make future QC decisions. 4 1 If possible, evaluate new QC material alongside existing QC material while determining a baseline mean for the new QC material. 2 Lab should use Within-lab standard deviation (SD) from assay sheet. Refer to "What Are?" for further information. 3 Refer to Definitions. Use the table that corresponds to the appropriate Performance Verifier product. 4 Source of QC procedures can be the "Statistical Tools of Quality Control: Trainining Module or your own QC procedures.. Flowchart 2. Establishing Baseline Means for Performance Verifiers VITROS Chemistry Systems 15

24 Using Monitoring System Performance on a Periodic (Monthly) Basis If you use as your daily control materials the results can be used to assess the performance of the VITROS Chemistry System on a periodic basis. This assessment might be done to detect drift or to verify a previous calibration. We recommend that this assessment be done on a monthly basis. Evaluate both the current (monthly) means and the calculated SDs. The monthly means are evaluated both for being within the ROM and whether or not the current mean is statistically the same as the baseline mean. The calculated standard deviations are evaluated for being larger than the PV assay sheet Within-lab SD. In order to evaluate whether the current mean is statistically different from the baseline mean, you need to calculate the difference (delta) between the current mean and the baseline mean. This difference is evaluated using the table in Appendix 2. The number of data points (n) included in this monthly assessment should match the number used to calculate the baseline mean (20-30). If the two (n)s are not the same, use the following guideline: If either the baseline mean or the current mean has n < 30 datapoints, use n = 20 from the table in Appendix 2. If both the baseline mean and the current mean have n 30 datapoints, use n = 30 from the table in Appendix 2. Use the steps outlined in Flowchart 3 to assess the performance of a VITROS Chemistry System on a periodic basis. In order to use this flowchart, you must use PVs as your daily control materials. Appendix 3 contains a worksheet that can be used to document the periodic monitoring of system performance. This procedure determines whether or not there is a statistically significant difference between your current mean and the baseline mean. A statistically significant difference may not be clinically significant. When you have a statistically significant difference, an investigation may be required, depending on your laboratory s action limit for monthly mean variation. 16 VITROS Chemistry Systems

25 Using Has a valid Baseline Mean 1 been established for PVs? No See Flowchart 5 Yes Has anything 2 happened which may have altered the performance of the VITROS Chemistry System? No Yes For each PV calculate the current mean 3 and the current standard deviation 3. You must evaluate both the mean and SD. See Flowchart 4 1 Refer to Definitions. 2 For example service or environmental changes 3 Refer to Definitions. 4 Use the delta value from Appendix 2. If either the baseline mean or the calculated monthly mean has n less than 30 data points, use n=20 from the Table. If both the baseline mean and the calculated monthly mean have n greater than or equal to 30 data points, use n=30 from the Table. Use the table that corresponds to the appropriate Performance Verifier product. 5 Some accreditation agencies and state regulations may require additional protocols. Comply as required. Are the current means 3 within the ROM? No Yes For each PV is the current SD 3 > Within-lab SD from PV assay sheet? Yes No For each PV is the difference between the current mean 3 and the Baseline Mean 1 the delta 4 in Appendix 2? Yes No Investigation may be required: Contact Technical Support Calibration is verified 5. There is no statistical evidence that the performance of the VITROS Chemistry System has changed. The current SD is within the expected range for a properly operating VITROS Chemistry System. Flowchart 3. Monitoring System Performance on a Periodic (Monthly) Basis VITROS Chemistry Systems 17

26 Using Verifying a Previous Calibration When System Performance May Have Changed should also be used to verify an existing calibration when the performance of the VITROS Chemistry System may have changed. The following situations may alter the performance of the VITROS Chemistry System to the extent that you should use Flowchart 4 to verify calibration: Major maintenance Environmental changes If you suspect that system performance has changed, use the mean results of duplicate measurements of fresh vials of to assess the current performance of the system. The calibration is still acceptable if the mean of two or more measurements of fresh vials of Performance Verifiers is within the ROM and these results are comparable to the baseline means. 18 VITROS Chemistry Systems

27 Using No Do you use PVs as your daily QC material? See Flowchart 5 Yes Run fresh vials Yes of PVs in duplicate. No Have valid Baseline Means 1 been established? Yes Yes For each PV calculate the average of these 2 values. Investigation required: Contact Technical Support No Are these means within the published Range of Means? Yes 1 Refer to Definitions. 2 Use the table that corresponds to the appropriate Performance Verifier product. 3 Some accreditation agencies and state regulations may require additional protocols. Comply as required. No For each PV is the difference between the 2-replicate average and the Baseline Mean 1 the delta 2 (for n=2) in Appendix 2? Yes Calibration is verified 3 Flowchart 4. Verifying A Previous Calibration When System Performance May Have Changed VITROS Chemistry Systems 19

28 Using Verifying a Previous Calibration When No Valid Baseline Mean for Has Been Established If you have not established a valid baseline mean for, and you need to verify a previous calibration, follow the steps outlined in Flowchart 5. There are several reasons for not having a valid baseline mean established. The two most common reasons are: You do not use as your daily QC material. There has not been enough time to establish the baseline mean (less than days). If you are verifying a previous calibration or you would like to assess the performance of the VITROS Chemistry System on a periodic basis and do not use PVs as a daily control you need to calculate the difference (delta) between your calibration verification mean (established at initial calibration) and today s 2-replicate mean on fresh vials of PVs. This difference is evaluated using the table in Appendix 2. If you are using PVs as your daily control and you need to verify a previous calibration but have not yet established a baseline mean, follow the steps outlined in Flowchart 5. You need to calculate the difference (delta) between your preliminary baseline mean (established at initial calibration) and today s 2-replicate mean on fresh vials of PVs. This difference is evaluated using the table in Appendix VITROS Chemistry Systems

29 Using Run fresh vials Yes of PVs in duplicate 1. For each PV calculate the average of these 2 values. Are these means within the published Range of Means? No Investigation required: Contact Technical Support Yes Recall the 2-replicate Calibration Verification Means or Preliminary Baseline Means. For each PV is the difference between today s 2-replicate average and the Calibration Verification Mean or Preliminary Baseline Mean the delta 2 (for n=2) in Appendix 2? Yes Calibration is verified 3 No 1 Must use the same lot number of PV now as was used to determine the inital mean. 2 Use the table that corresponds to the appropriate Performance Verifier product. 3 Some accreditation agencies and state regulations may require additional protocols. Comply as required. Flowchart 5. Verifying a Previous Calibration When No Valid Baseline Mean for (PVs) Has Been Established VITROS Chemistry Systems 21

30 Using 22 VITROS Chemistry Systems

31 Appendix 1: Performance Verifier Assay Sheets This Appendix contains a sample of the Performance Verifier assay sheets VITROS Chemistry System 23

32 Appendix 1: Performance Verifier Assay Sheets VITROS Performance Verifier II P AcP ALB ALKP ALT AMYL AST Bc Bu GEN U/L g/dl U/L U/L U/L U/L mg/dl mg/dl U/L g/l U/L U/L U/L U/L µmol/l µmol/l VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. 1 (9) 24 VITROS Chemistry System

33 Appendix 2: Tables The Tables in this Appendix are used to evaluate system drift or performance shifts. Data in the tables were developed using the within-lab SD found on the Performance Verifier assay sheets. Table 1. VITROS I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 AcP 1 U/L U/L AcP 2 U/L U/L ALB 1 g/dl g/l ALB 2 g/dl g/l ALKP 1 U/L U/L ALKP 2 U/L U/L ALT 1 U/L U/L ALT 2 U/L U/L AMYL 1 U/L U/L AMYL 2 U/L U/L AST 1 U/L U/L AST 2 U/L U/L Bc 1 mg/dl µmol/l Bc 2 mg/dl µmol/l VITROS Chemistry Systems 25

34 Appendix 2: Tables Chemistry PV Level Units Table 1. VITROS I and II (Continued) Conventional Units SI Units n=2 n=20 n=30 Units Bu 1 mg/dl µmol/l Bu 2 mg/dl µmol/l BUN 1 mg/dl mmol/l BUN 2 mg/dl mmol/l Ca 1 mg/dl mmol/l Ca 2 mg/dl mmol/l CHE 1 U/mL U/L CHE 2 U/mL U/L CHOL 1 mg/dl mmol/l CHOL 2 mg/dl mmol/l CK 1 U/L U/L CK 2 U/L U/L Cl - 1 mmol/l mmol/l Cl - 2 mmol/l mmol/l CREA 1 mg/dl µmol/l CREA 2 mg/dl µmol/l DGXN 1 ng/ml nmol/l DGXN 2 ng/ml nmol/l dhdlc 1 mg/dl mmol/l dhdlc 2 mg/dl mmol/l dldl 1 mg/dl mmol/l dldl 2 mg/dl mmol/l ECO 2 1 mmol/l mmol/l ECO 2 2 mmol/l mmol/l Fe 1 µg/dl µmol/l Fe 2 µg/dl µmol/l GGT 1 U/L U/L GGT 2 U/L U/L GLU 1 mg/dl mmol/l GLU 2 mg/dl mmol/l HDLC 1 mg/dl mmol/l HDLC 2 mg/dl mmol/l K + 1 mmol/l mmol/l K + 2 mmol/l mmol/l LAC 1 mmol/l mmol/l LAC 2 mmol/l mmol/l n=2 n=20 n=30 26 VITROS Chemistry Systems

35 Appendix 2: Tables Chemistry PV Level Units Table 1. VITROS I and II (Continued) Conventional Units SI Units n=2 n=20 n=30 Units LDH 1 U/L U/L LDH 2 U/L U/L Li 1 mmol/l mmol/l Li 2 mmol/l mmol/l LIPA 1 U/L U/L LIPA 2 U/L U/L Mg 1 mg/dl mmol/l Mg 2 mg/dl mmol/l Na + 1 mmol/l mmol/l Na + 2 mmol/l mmol/l PHOS 1 mg/dl mmol/l PHOS 2 mg/dl mmol/l PHYT 1 µg/ml µmol/l PHYT 2 µg/ml µmol/l TBIL 1 mg/dl µmol/l TBIL 2 mg/dl µmol/l THEO 1 µg/ml µmol/l THEO 2 µg/ml µmol/l TIBC 1 µg/dl µmol/l TIBC 2 µg/dl µmol/l TP 1 g/dl g/l TP 2 g/dl g/l TRIG 1 mg/dl mmol/l TRIG 2 mg/dl mmol/l URIC 1 mg/dl µmol/l URIC 2 mg/dl µmol/l n=2 n=20 n= VITROS Chemistry Systems 27

36 Appendix 2: Tables Table 2. VITROS Liquid I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 ALC 1 mg/dl mmol/l ALC 2 mg/dl mmol/l AMON 1 µmol/l µmol/l AMON 2 µmol/l µmol/l GLU (CSF) 1 mg/dl mmol/l GLU (CSF) 2 mg/dl mmol/l K + (ur) 1 mmol/l mmol/l K + (ur) 2 mmol/l mmol/l Na + (ur) 1 mmol/l mmol/l Na + (ur) 2 mmol/l mmol/l PROT (CSF) 1 mg/dl mg/l PROT (CSF) 2 mg/dl mg/l SALI 1 mg/dl mmol/l SALI 2 mg/dl mmol/l Table 3. VITROS Isoenzyme I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 CKMB 1 U/L U/L CKMB 2 U/L U/L CK 1 U/L U/L CK 2 U/L U/L VITROS Chemistry Systems

37 Appendix 2: Tables Table 4. VITROS TDM I, II and III Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 ACET 1 µg/ml µmol/l ACET 2 µg/ml µmol/l ACET 3 µg/ml µmol/l CAFFN 1 µg/ml µmol/l CAFFN 2 µg/ml µmol/l CAFFN 3 µg/ml µmol/l CRBM 1 µg/ml µmol/l CRBM 2 µg/ml µmol/l CRBM 3 µg/ml µmol/l DGXN 1 µg/ml µmol/l DGXN 2 µg/ml µmol/l DGXN 3 µg/ml µmol/l GENT 1 µg/ml µmol/l GENT 2 µg/ml µmol/l GENT 3 µg/ml µmol/l PHBR 1 µg/ml µmol/l PHBR 2 µg/ml µmol/l PHBR 3 µg/ml µmol/l PHYT 1 µg/ml µmol/l PHYT 2 µg/ml µmol/l PHYT 3 µg/ml µmol/l TOBRA 1 µg/ml µmol/l TOBRA 2 µg/ml µmol/l TOBRA 3 µg/ml µmol/l VALP 1 µg/ml µmol/l VALP 2 µg/ml µmol/l VALP 3 µg/ml µmol/l VANC 1 µg/ml µmol/l VANC 2 µg/ml µmol/l VANC 3 µg/ml µmol/l VITROS Chemistry Systems 29

38 Appendix 2: Tables Table 5. VITROS CRP I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 CRP 1 mg/dl mg/l CRP 2 mg/dl mg/l Table 6. VITROS UPRO I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 UPRO 1 mg/dl g/l UPRO 2 mg/dl g/l Table 7. VITROS AAT/HPT I, II and III Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 AAT 1 mg/dl g/l AAT 2 mg/dl g/l AAT 3 mg/dl g/l HPT 1 mg/dl g/l HPT 2 mg/dl g/l HPT 3 mg/dl g/l Table 8. VITROS dtibc I and II Conventional Units Table 9. VITROS HCY I, II, and III SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 dtibc 1 ug/dl umol/l dtibc 2 ug/dl umol/l Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 HCY 1 umol/l umol/l HCY 2 umol/l umol/l HCY 3 umol/l umol/l VITROS Chemistry Systems

39 Appendix 2: Tables Table 10. VITROS ApoA1 Performance Verifier I Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 ApoA1 1 mg/dl g/l Table 11. VITROS ApoB Performance Verifier I Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 ApoB 1 mg/dl g/l Table 12. VITROS ASO/RF I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 ASO 1 IU/mL IU/mL ASO 2 IU/mL IU/mL RF 1 IU/mL IU/mL RF 2 IU/mL IU/mL Table 13. VITROS hscrp I, II and III Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 hscrp 1 mg/dl mg/l hscrp 2 mg/dl mg/l hscrp 3 mg/dl mg/l Table 14. VITROS malb I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 malb 1 mg/dl mg/l malb 2 mg/dl mg/l VITROS Chemistry Systems 31

40 Appendix 2: Tables Table 15. VITROS PALB Performance Verifier I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 PALB 1 mg/dl mg/l PALB 2 mg/dl mg/l Table 16. VITROS Protein I, II and III Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 C3 1 mg/dl mg/l C3 2 mg/dl mg/l C3 3 mg/dl mg/l C4 1 mg/dl mg/l C4 2 mg/dl mg/l C4 3 mg/dl mg/l IgA 1 mg/dl g/l IgA 2 mg/dl g/l IgA 3 mg/dl g/l IgG 1 mg/dl g/l IgG 2 mg/dl g/l IgG 3 mg/dl g/l IgM 1 mg/dl g/l IgM 2 mg/dl g/l IgM 3 mg/dl g/l TRFRN 1 mg/dl g/l TRFRN 2 mg/dl g/l TRFRN 3 mg/dl g/l VITROS Chemistry Systems

41 Appendix 2: Tables Table 17. VITROS %A1c I and II Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 %A1c (IFCC) 1 % % %A1c (IFCC) 2 % % %A1c (NGSP) 1 % % %A1c (NGSP) 2 % % Hb 1 g/dl g/l Hb 2 g/dl g/l HbA1c 1 g/dl g/l HbA1c 2 g/dl g/l VITROS Chemistry Systems 33

42 Appendix 2: Tables Table 18. VITROS DAT I, II, III, IV, and V Conventional Units SI Units Chemistry PV Level Units n=2 n=20 n=30 Units n=2 n=20 n=30 AMPH 1 ng/ml ug/l AMPH 2 ng/ml ug/l AMPH 3 ng/ml ug/l AMPH 4 ng/ml ug/l AMPH 5 ng/ml ug/l BARB 1 ng/ml ug/l BARB 2 ng/ml ug/l BARB 3 ng/ml ug/l BARB 4 ng/ml ug/l BARB 5 ng/ml ug/l BENZ 1 ng/ml ug/l BENZ 2 ng/ml ug/l BENZ 3 ng/ml ug/l BENZ 4 ng/ml ug/l BENZ 5 ng/ml ug/l COCM 1 ng/ml ug/l COCM 2 ng/ml ug/l COCM 3 ng/ml ug/l COCM 4 ng/ml ug/l COCM 5 ng/ml ug/l METD 1 ng/ml ug/l METD 2 ng/ml ug/l METD 3 ng/ml ug/l METD 4 ng/ml ug/l METD 5 ng/ml ug/l OP 1 ng/ml ug/l OP 2 ng/ml ug/l OP 3 ng/ml ug/l OP 4 ng/ml ug/l OP 5 ng/ml ug/l PCP 1 ng/ml ug/l PCP 2 ng/ml ug/l PCP 3 ng/ml ug/l THC 1 ng/ml ug/l THC 2 ng/ml ug/l VITROS Chemistry Systems

43 Appendix 2: Tables Chemistry PV Level Units Conventional Units SI Units n=2 n=20 n=30 Units THC 3 ng/ml ug/l THC 4 ng/ml ug/l n=2 n=20 n= VITROS Chemistry Systems 35

44 Appendix 2: Tables 36 VITROS Chemistry Systems

45 Appendix 3: Quality Control Worksheets This Appendix contains the following Quality Control worksheets: Calibration Verification Baseline Mean Evaluation Periodic Monitoring of System Performance VITROS Chemistry Systems 37

46 Appendix 3: Quality Control Worksheets VITROS Chemistry System QC Worksheet Calibration Verification (Preliminary Baseline Mean) Chemistry: Body Fluid: Performance Verifier I Lot #: Performance Verifier II Lot #: Performance Verifier III Lot #: Expires: Expires: Expires: MicroSlide/MicroTip Reagent Lot #: NOTE: MicroSlide Lot is 12 digits MicroTip Reagent Lot is 10 digits Expires: Date of Calibration: Calibrated by: Performance Verifier I Performance Verifier II ROM: Is the Average within ROM: Is the Average within the ROM? the ROM? Replicate 1: Yes No Replicate 2: Average: Replicate 1: Yes No Replicate 2: Average: Performance Verifier III ROM: Is the Average within the ROM? Attach the calibration report printout to this worksheet for future reference. Replicate 1: Yes No Replicate 2: Average: Check the appropriate box If the average for each Performance Verifier is within its ROM, then CALIBRATION IS VERIFIED. If the average of any Performance Verifier is NOT within its ROM, then CALIBRATION IS NOT VERIFIED. Reviewed by: Date: 38 VITROS Chemistry Systems

47 Appendix 3: Quality Control Worksheets VITROS Chemistry System QC Worksheet Baseline Mean Evaluation Chemistry: Body Fluid: MicroSlide/MicroTip Reagent Lot #: Expires: NOTE: MicroSlide Lot is 12 digits. MicroTip Reagent Lot is 10 digits. Date of Calibration: Performance Verifier I, Lot # ROM: Value*: Baseline Mean: Is the Baseline Mean within the ROM? Preliminary Baseline Mean** Is the difference between the Baseline Mean and the Preliminary Baseline mean less than or equal to the Calculated *? Yes No Value Yes No N=: Performance Verifier II, Lot # ROM: Value*: Baseline Mean: Is the Baseline Mean within the ROM? Yes No Preliminary Baseline Mean** Is the difference between the Baseline Mean and the Preliminary Baseline mean less than or equal to the Calculated *? Value Yes No N=: Performance Verifier III, Lot # ROM: Value*: Baseline Mean: Is the Baseline Mean within the ROM? Preliminary Baseline Mean** Is the difference between the Baseline Mean and the Preliminary Baseline mean less than or equal to the Calculated *? Yes No Value Yes No N=: *Use the Calculated for n=2 to verify that the differences between the Preliminary Baseline Means and the Baseline Means are not significant. The Calculated is equal to the absolute value of the difference between the current mean and the baseline mean. ** The Preliminary Baseline Mean is recorded in the Calibration Verification Worksheet Check the appropriate box If the answers to all questions were YES, then the Baseline means are acceptable. If the answer to any question was NO, then an investigation is required. Reviewed by: Date: VITROS Chemistry Systems 39

48 Appendix 3: Quality Control Worksheets VITROS Chemistry System QC Worksheet For Periodic Monitoring of System Performance Chemistry: Body Fluid: Performance Verifier Lot #: Expires: MicroSlide/MicroTipReagent Lot # Expires Value**, n = 2: Baseline Mean: NOTE: MicroSlide Lot is 12 digits. MicroTip Reagent Lot is 10 digits. Value**, n = 20 Baseline SD***: Date of Initial Calibration: Value**, n = 30: ROM: Is the Is * < or Is Current Reporting Period Current Mean equal to SD > the Current In the ROM? Calculated Value**? Current Baseline SD? Begin End Mean n Yes No * Yes No SD Yes No Corrective Action**** **** If any of the questions have a check mark in shaded columns, corrective action may be required. * The Calculated is equal to the absolute value of the difference between the current mean and the baseline mean. current mean - baseline mean ** Use the tables in Appendix 2 of the Performance Verifier Training Module for the Values on the appropriate PV fluids. *** Use the Within-Lab SDs from the Performance Verifier assay sheets as Baseline SDs. Reviewed by: Date: 40 VITROS Chemistry Systems