liquicolor Design Verification

Transcription

1 Design Verification liquicolor 1 Introduction Imprecision Imprecision for Imprecision for Linearity and Detection Limit Linearity Detection Limit Recovery in Sera Comparison of Methods Stability Data Accelerated Stress Tests Real Time Stability Interferences Traceability... 9 QM-Element: ISO9001:2000 No /9 valid of

2 1 Introduction The performance characteristics of + T liquicolor have been tested and documented in order to verify the clinical usefulness and compliance with the essential requirements of directive 98/79/EC. 2 Imprecision The imprecision (within-run and day-to-day) of + T liquicolor was calculated from six determinations on ten consecutive days. sera with low, medium and high bilirubin levels were employed as sample material. 2.1 Imprecision for Analyte concentration Intra-assay Inter-assay () SD () CV SD () CV Imprecision for Analyte concentration Intra-assay Inter-assay () SD () CV SD () CV Linearity and Detection Limit 3.1 Linearity The linearity of + T liquicolor was controlled by employing a high concentration pool and dilutions with physiological saline. The analysed concentrations were calculated vs. the regression line. from the regression line are expressed in absolute and relative values. As an example data measured on the Hitachi 717 are reported. Hitachi 717 High Pool Analytical Data Regressed Data from Regression Line Content Design Verification and Product Data for + T liquicolor 2/9

3 MEASURED ANALYTE CONCENTRATION () 60 LINEARITY Linear Regression 50 Error Bars = ± 2SD HIGH POOL CONTENT High Pool Analytical Data Regressed Data from Regression Line Content Design Verification and Product Data for + T liquicolor 3/9

4 MEASURED ANALYTE CONCENTRATION () 30 LINEARITY Linear Regression 25 Error Bars = ± 2SD HIGH POOL CONTENT + T liquicolor has a linear range for both direct and total bilirubin up to Detection Limit A 20-fold determination of a 0 sample (phys. saline) on a Hitachi 717 analyser revealed an absolute mean of 0.00 and a SD of 0.01 (total bilirubin) and an absolute mean of 0.01 with SD of 0.08 (direct bilirubin). From the three-fold standard deviation the detection limit can therefore be calculated on the base of mean +3 SD to 0.03 total bilirubin and to 0.25 bilirubin direct. 4 Recovery in Sera Commercially available control sera have been employed. The control sera have been reconstituted/prepared according to the manufacturer s instructions. Fivefold determinations of each control have been performed with + T liquicolor reagents. The means of the fivefold determinations have been calculated and compared with the target values. recovery Reference liquicolor Target Range HumaTrol N # HumaTrol P # SERODOS # SERODOS plus # Precinorm # Precipath # Sum Mean Design Verification and Product Data for + T liquicolor 4/9

5 recovery Reference liquicolor Target Range HumaTrol N # HumaTrol P # SERODOS plus # Precinorm # Precipath # Sum Mean With the exception of Precipath all control sera were recovered within the specified ranges. It should be noted however, that the target value for this control was not established for + T liquicolor method. There was a good agreement observed of the reference test and BILIRUBIN liquicolor D + T in regard to the summary of recoveries and mean deviations. 5. Comparison of Methods + T liquicolor has been compared against commercially available bilirubin methods. sera as well as patient samples have been employed in the comparison. The results have been evaluated by a main component analysis. The linear regressions obtained could be described as follows: BILIRUBIN direct N = 52 Y = X r = ,0 Method Comparison Bilirubin D liquicolor 4,5 4,0 BILIRUBIN_T_LIQUICOLOR, 3,5 3,0 2,5 2,0 1,5 1,0 0,5 0,0 0,0 0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 REF., Design Verification and Product Data for + T liquicolor 5/9

6 BILIRUBIN total N = 57 Y = X r = ,5 Method Comparison Bilirubin T liquicolor 2,0 liquicolor 1,5 1,0 0,5 0,0 0,0 0,5 1,0 1,5 2,0 2,5 REF., The methods showed a very good agreement and no significant deviation could be observed. 6. Stability Data 6.1 Accelerated Stress Tests Recovery in Sera Commercially available control sera have been employed. The control sera have been reconstituted/prepared according to the manufacturer s instructions. Fivefold determinations of each control have been performed with + T liquicolor method after 10 and 12 days storage at 37 C. The means of the fivefold determinations have been calculated and compared with the values of fresh reagent. BILIRUBIN direct CONTROL SERUM RECOVERY Reagent fresh Reagent 10 days 37 C Reagent 12 days 37 C HumaTrol N # HumaTrol P # SERODOS # SERODOS plus # Precinorm # Precipath # Mean Design Verification and Product Data for + T liquicolor 6/9

7 CONTROL SERUM RECOVERY Reagent fresh Reagent 10 days 56 C Reagent 14 days 56 C HumaTrol N # HumaTrol P # SERODOS # SERODOS plus # Precinorm # Precipath # Mean Linearity The linearity of + T liquicolor was controlled according to the following procedure control sera have been reconstituted/prepared according to the manufacturer s instructions. Twofold determinations of each control have been performed. The reagents were stressed 10 and 14 days at 56 C. High Pool Content Reagent Reagent 10 days 56 C 14 days 56 C Analytical Regressed from Analytical Regressed from Data Data Regression Line Data Data Regression Line High Pool Content Reagent Reagent 10 days 56 C 14 days 56 C Analytical Regressed from Analytical Regressed from Data Data Regression Line Data Data Regression Line Design Verification and Product Data for + T liquicolor 7/9

8 6.2 Real Time Stability The stability of + T liquicolor has been tested on real-time storage conditions over a period of 24 and 28 months for two independent production lots liquicolor (H033, exp. date: February 2004; H038, exp. date: July 2004;), and over a period of 24, 25, 28 and 30 months for four independent production lots liquicolor (H025, exp. date: December 2003; H033, exp. date: February 2004; H035, exp. date: May 2004; H038, exp. date: July 2004;). Measurements were made on Hitachi 717. Acceptance criteria were based on the recovery of controls. Twofold determinations of each control have been performed with and T liquicolor. The means of the twofold determinations have been calculated and compared with the values of the reference reagent. CONTROL fresh (Refer) H months H month HumaTrol N # HumaTrol P # SERODOS # SERO.plus # Mean CONTROL BILIRUBIN T fresh (Refer) H months H months H months H 033 H months H 025 HumaTrol N # HumaTrol P # SERODOS # SERO.plus # Mean All results support the claimed stability of 24 months from production for the test kit. 7. Interferences Interferences by lipemia (intralipid), hemoglobin and ascorbic acid have been studied by adding known amounts of the potentially interfering substances to a known sample. Recoveries have been analyzed according to the method of Glick et al. (Clin.Chem. 1986, ). Criterion for significance: recovery within 10 of original. Design Verification and Product Data for + T liquicolor 8/9

9 Intralipid.,, Hemo.,, Ascorb. acid,, Triglycer., Glick Intralipid.,, Hemo.,, Ascorb. acid,, Triglycer., Glick The test is not significantly interfered with by intralipid < 400. Hemolytic sera below 50 hemoglobin, ascorbic acid below 2 and lipemic sera (triglycerides below 250 ) do not significantly interfere with the test. The test is not significantly interfered with by intralipid < 600, and triglycerides < 750. Hemolytic samples > 50 and ascorbic acid above 2 interfere with the test. 8. Traceability Currently there is no agreed international reference preparation for direct bilirubin available. The traceability has therefore been documented by a comparison of methods (see 4). The total bilirubin concentration has been verified with the reference material SRM 909b. The total bilirubin recovery was found well within the reproducibility limits of the method, therefore the method is traceable to SRM 909b. Design Verification and Product Data for + T liquicolor 9/9