PEPFAR Quality Control and Method Validation Activity 14 Handout 4: AST Validation Summary

Handout 4: AST Validation Summary 1406 1 of 6 Validation Summary Report Purpose: Validation Re-Validation X Other: Verification of Manufacturer’s Claims Description of Equipment/Process: Equipment/Process: Chemistry Analyzer regarding AST only Serial Number: 12345 Location: Cape Clinic Laboratory, 111 Main Street, Arusha, Tanzania Date: November 1 through 30, 2013 FDA Approval Status: X Approved Not approved Procedure: Refer to the Cape Clinic Laboratory Validation Plan regarding AST Results: All raw data reports and statistical analysis can be found in the Validation binder. 1. Precision -- refer to tab A Observed Results Expected Results Analyte Between Day Acceptability Manufacturer’s Normal Control Precision 33% of CLIA Abn Control CV% CV% Ex:ALT 3.3% 6.6% 3.8% 4.3% Acceptable AST 1.3% 6.6% Observed Results Expected Results Analyte Within Run Acceptability Manufacturer’s Normal Control Abn Control Precision 25% of CLIA CV% CV% Ex:ALT 2.6% 5% 2.6% 4.3% Acceptable AST 0.8% 5% Activity 14: Introduction to Method Evaluation Modified from pSMILE.org website. Equ35-A-02_Chem_Val_Sum_Template.Last accessed May 3, 2013 HILS1771 Effective Date: 12/06/2016 Handout 4: AST Validation Summary (1406) 2 of 6 2. Accuracy -- refer to tab B Correlation with Cape Clinic chemistry department’s XYZ Chemistry Analyzer using forty patient specimens spanning the AMR for AST will be analyzed in duplicate on each instrument. At least twenty of the specimens selected must be outside of the current reference interval for AST. Specimens selected will be analyzed on the same day of collection and within two hours between each instrument. Replicates will be analyzed in different runs and then reviewed for any significant discrepancies, such as sample mix-ups or transposition errors. Patient samples will be collected and handled in accordance with Cape Clinic’s collection and handling procedures. A serum aliquot from a centrifuged red top collection tube will be analyzed. Sufficient quantities of each sample (minimum 1.0 ml serum) must be available to: 1-) analyze duplicates by the test methods, 2- ) analyze duplicates by the comparative methods, and 3-) perform follow-up studies if needed. Data collection will occur over a span of 10 working days, analyzing four specimens per day. Total Correlation Linear Regression Acceptability Analyte Coefficient (r) Allowable Statistics For Linear Error Expected Slope Intercept Regression >0.975 Ex.ALT 0.5U/L or 20% 0.997 1.002 2.19 Acceptable AST 20% Analyte Xc Linear Regression Equation Bias at Xc %Bias at Xc Ex.ALT 40 U/L Y= 1.002 X + 2.19 2.3 U/L 5.7% AST 35 U/L AST 120 U/L Activity 14: Introduction to Method Evaluation HILS1771 Modified from pSMILE.org website. Equ35-A-02_Chem_Val_Sum_Template.Last accessed May 3, 2013 Effective Date: 12/06/2016 Handout 4: AST Validation Summary (1406) 3 of 6 3. Linearity -- refer to tab C i. Linearity verification will be determined using patient samples previously analyzed on the XYZ analyzer. A known high near or slightly above the upper measurement range and a known low near the limit of detection will be used. Linear Regression Allowable Linear Analyte Statistics Systematic Error Range Evaluation Slope Intercept 50% of TEa Verified Ex:ALT 0.970 0.282 10% 2.5-770 Linear 4. Analytical Measurement Range (AMR) and Clinical Reportable Range (CRR) -- refer to tab C Low High Analyte Mfg’s AMR Value Value Reportable Verified Verified Range Dilutions CRR Ex:ALT 5-700 U/L 2.5 770 5-700 1:10 5-7000 5. Sensitivity and Specificity -- refer to tab D for instrument package inserts Summary of Manufacturer’s Claims for Sensitivity and Specificity Analyte Specificity (Interfering Substances) Sensitivity AST Icterus – No significant effect 5 U/L Hemolysis – No significant effect Lipemia – No significant effect; >Abs flagging may occur Drugs – No interference at therapeutic concentrations; Isoniazid may cause artificially low and Furosemide may cause artificially high AST results at therapeutic concentrations Activity 14: Introduction to Method Evaluation Modified from pSMILE.org website. Equ35-A-02_Chem_Val_Sum_Template.Last accessed May 3, 2013 HILS1771 Effective Date: 12/06/2016 Handout 4: AST Validation Summary (1406) 4 of 6 5. Reference ranges -- refer to tab E a. The reference range studies have been verified for the following populations: i. Adult reference ranges –Reference ranges for males and females were transferred from the XYZ manufacturer’s expected values. The transference was verified using forty serum samples (twenty adult males and twenty adult females) from Cape Clinic’s employee population. ii. Pediatric reference ranges– Reference ranges for pediatrics were adopted from Ugandan reference ranges in use at the XYZ Research lab in Kampala, Uganda. The ranges were evaluated by the Cape Clinic medical leadership team in consultation with local pediatricians and determined to be appropriate for the local Tanzanian population. Adoption of these ranges was approved by the Cape Clinic Medical Director. All ranges will be verified and monitored over time as appropriate data becomes available. Analyte Adult Reference Ranges Reference Range Cited % Verified (Expected ≥90%) Males ≤ 38 U/L XYZ Manufacturer Range AST Females ≤32 U/L Reference Range Approval Medical Director: _______________________________ Date: _______________ Insert Medical director name here Activity 14: Introduction to Method Evaluation Modified from pSMILE.org website. Equ35-A-02_Chem_Val_Sum_Template.Last accessed May 3, 2013 HILS1771 Effective Date: 12/06/2016 Handout 4: AST Validation Summary (1406) 5 of 6 Acceptability of Method The Chemistry Analyzer may be judged acceptable for the AST analyte if one of the following conditions is met 1. Manufacturer’s claims for linearity, precision and accuracy have been verified 2. The total error calculation (bias + 3 sd or %bias + 3 %CV) for the test method is less than the CLIA total allowable error for each Medical Decision Point (Xc). 3. The Sigma-metric is > 3.0. Concentration AST %TEA at Long- %Bias or Total Sigma Acceptability of Xc concentration term %difference Error Metric of Xc Precision with (in %) (%CV) comparative method Activity 14: Introduction to Method Evaluation Modified from pSMILE.org website. Equ35-A-02_Chem_Val_Sum_Template.Last accessed May 3, 2013 HILS1771 Effective Date: 12/06/2016 Handout 4: AST Validation Summary (1406) 6 of 6 Method Approval Approved / Not Approved If not approved, provide recommendations/corrective actions below. Laboratory Director: ____________________________ Date: _________________ Insert Lab Director name here Prepared by: ______________________________ Date: ___________________ Insert name and title here Activity 14: Introduction to Method Evaluation Modified from pSMILE.org website. Equ35-A-02_Chem_Val_Sum_Template.Last accessed May 3, 2013 HILS1771 Effective Date: 12/06/2016