Technical Bulletin. Performance Characteristics of the BN ProSpecä System. By Liana Frances Romero, BS, MT(ASCP)

Transcription

1 Technical Bulletin Performance Characteristics of the BN ProSpecä System By Liana Frances Romero, BS, MT(ASCP) Introduction The BN ProSpecä System is an automated nephelometry analyzer for in vitro diagnostic use in clinical laboratories. The BN ProSpecä System is intended to determine the concentration of antigen-antibody complexes in a suspension by measuring their light scattering properties. The BN ProSpecä System is used in conjunction with Dade Behring reagents to measure the concentration of a variety of proteins in serum, plasma, urine and CSF. The performance characteristics of the BN ProSpecä System were documented via studies conducted in the field as well as internally at Dade Behring. External studies were performed at the following sixteen sites throughout Europe, Canada and the United States: Hospital Jean Verdier Bondy, France Azienda Ospedaliera di Bologna Bologna, Italy Groupe Hospitalier DASMELAB Università di Napoli - Pitié Salpètrierè Federico II Paris, Frances Napoli, Italy ` Hospital General Yagùe Burgos, Spain Hospital Universitario de Getafe Getafe, Madrid, Spain rdwestkrankenhaus Frankfurt, Germany Laboratorium Dr. Merten & Partner Köln-Lindenthal, Germany Freie Universität Berlin Berlin, Germany Ospedale Martini Nuovo Torino, Italy Hospital for Sick Children Toronto, Ontario, Canada Boston Children s Hospital Boston, MA, USA Foundation for Blood Research Scarborough, ME, USA Hospital for Joint Diseases New York, NY, USA University Medical Center SUNY at Stony Brook, NY, USA University of Maryland, Baltimore Baltimore, MD, USA 1

2 Studies performed included calibration stability, reproducibility, method comparison, interference testing, and linearity. te: Data generated from these studies represent typical performance of the BN ProSpecä System in an average laboratory setting and should not be interpreted as specifications. Product Performance Calibration Stability The following assays were evaluated for calibration stability: Table 1 Analytes Evaluated for Calibration Stability N Antiserum to Human IgG (IgG) N Latex High Sensitivity CRP (CRP2) N Antiserum to Human IgG/Urine (IgGU)* N Antiserum to Human C1-Inhibitor (C1I) N Antiserum to Human IgA (IgA) N Antiserum to Human Plasminogen (PLS) N Antiserum to Human IgM (IgM) N Latex Myoglobin (MYO) N Antiserum to Human C3c (C3) N Latex ASL (ASL) N Antiserum to Human C4 (C4) N Latex ADNase B (ADNs) N Antiserum to Human Transferrin (TRF) N Latex Cystatin C N Antiserum to Human Transferrin/Urine (TRFU) N Antiserum to Human Fibronectin (FNC)* N Antiserum to Human Albumin (ALB) N Latex Lp(a) Reagent (LPa)* N Antiserum to Human Albumin/Urine (ALBU) Total Protein (20% TCA) (TPS) N Antiserum to Human Albumin/CSF (ALBC) Total Protein (20% TCA)/Urine (TPU)* N Antiserum to Human α1-antitrypsin (AAT) Total Protein (20% TCA)/CSF (TPC)* N Antiserum to α2-macroglobulin (α2m) N Antiserum to Human Ig/L-Chain, K-Type (kap) N Antiserum to Human Haptoglobin (HPT) N Antiserum to Human Ig/L-Chain, L-Type (lam) N Antiserum to Human α 1 acidglycoprotein(aag) N Antiserum to Human Ig/L-Chain, K-Type/ Urine (kapu)* N Antiserum to Human Prealbumin (PRE) N Antiserum to Human Ig/L-Chain, L-Type/ Urine (lamu)* N Latex stfr (stfr) N Latex IgE mono (IgE) N Antiserum to Human Ceruloplasmin (CER) N Antiserum to Human IgG1-4/(IgG1-4) N Antiserum to Human Retinol-binding Protein (RBP) N Antiserum to Human IgA (Pediatric) (IgAp) N Antiserum to Human Apolipoprotein A-I (A_I) N Antiserum to Human IgM (Pediatric) (IgMp) N Antiserum to Human Apolipoprotein B (ApoB) N Antiserum to Human IgA (CSF) (IgAC) N Antiserum to Human Apolipoprotein E (ApoE)* N Antiserum to Human IgG (CSF) (IgGC) N Antiserum to Human Apolipoprotein A-II (A_II)* N Antiserum to Human IgM (CSF) (IgMC)* N Antiserum to Human Fibrinogen (FIB) N alpha1 Microglobulin (α1mu) N Antiserum to Human Antithrombin III (AT3) N Antiserum to Human alpha2-macroglobulin (α2m) N Latex RF (RF) N Antiserum to Human alpha2-macroglobulin/ Urine (α2mu) N Latex Ferritin (FRT) N Antiserum to Human alpha2-macroglobulin/ CSF (α2mc) (*These assays are currently not available in the United States) 2

3 All assays to be evaluated at a specific site were calibrated at the beginning of the study. The stability of the reference curves was monitored using a quality control (QC) program. The data obtained demonstrated that the calibrations remained valid for the entire study period (an average of 8 weeks) at all sites. Reproducibility Reproducibility testing was conducted for the assays listed in Table 2 using each level of the corresponding Dade Behring control product. The control products were tested twice per day, in replicates of four for five days, to total forty (40) replicates. Table 2 BN ProSpec System Summary of Precision Studies Unit n Control Level Mean Within- Run Between- Run Total 1 N Antiserum to Human IgG N Antiserum to Human IgA N Antiserum to Human IgM N Antiserum to Human C3c N Antiserum to Human C4 N/T Prot.Control SL-L N Antiserum to Human Transferrin/Serum N Antiserum to Human Albumin N Antiserum to Human α1- Antitrypsin N Antiserum to Human α2- Macroglobulin N Antiserum to Human Haptoglobin N Antiserum to Human α1-acid Glycoprotein

4 Table 2 (Cont.) BN ProSpec System Summary of Precision Studies Unit n Control Level Mean Within- Run Between- Run Total 12 N Antiserum to Human Prealbumin N Latex stfr mg/l N Antiserum to Human Ceruloplasmin N Antiserum to Human Retinol binding Protein 17 N Antiserum to Human Ig/Lchain, kappa N Antiserum to Human Ig/Lchain, lambda N Antiserum to Human Transferrin/Urine mg/l 40 N/T Prot. Control LC Urine Pool L Urine Pool H N Antiserum to Human Albumin/Urine mg/ml N Antiserum to Human α1 Microglobulin/Urine mg/ml 40 N/T Prot. Control LC Urine Pool L Urine Pool H N/T Prot. Control LC Urine Pool L Urine Pool H N Antiserum to Human Apolipoprotein A-I Apolipoprotein Control - Serum CHD Serum Pool L Serum Pool H N Antiserum to Human Apolipoprotein B Apolipoprotein Control - Serum CHD Serum Pool L Serum Pool H N Antiserum to Human Fibrinogen N/T Prot. Control PY Plasma Pool L Plasma Pool H N Antiserum to Human Antithrombin III N/T Prot. Control PY Plasma Pool L Plasma Pool H N High Sensitivity CRP mg/l 40 Apolipoprotein Control - Serum CHD Serum Pool L Serum Pool H

5 Table 2 (Cont.) BN ProSpec System Summary of Precision Studies Unit n Control Level Mean Within- Run Between- Run Total 43 N Antiserum to Human Plasminogen N/T Prot. Control PY Plasma Pool L Plasma Pool H N Latex Ferritin µg/l N Latex RF IU/mL 40 N/T Rheum. Contr. SL/1 N/T Rheum. Contr. SL/ N Latex ASL IU/mL 40 N/T Rheum. Contr. SL/1 N/T Rheum. Contr. SL/ N Latex IgE IU/mL N Latex ADNase B U/mL 40 NADNaseBControl N/T Rheum. Contr. SL/ N Latex Myoglobin µg/l 40 N Myoglobin Control Serum Pool L Serum Pool H N Antiserum to Human IgG/CSF mg/l 40 N/T Prot. Control LC CSF Pool L CSF Pool H N Latex IgA/CSF mg/l 40 N/T Prot. Control LC N IgA Control CSF Pool L CSF Pool H N Antiserum to Human C1 Inhibitor N/T Prot. Control PY Serum Pool L Serum Pool H N High Sensitivity CRP mg/l 40 N/T Rheum. Contr. SL/1 N/T Rheum. Contr. SL/

6 Method Comparison Correlation studies were conducted for the assays listed in Table 3 on the BN ProSpecä System versus the BN IIä System. Samples were selected to span the reporting range of each assay evaluated in this study. Regression statistics (correlation coefficient, slope, and y-intercept) were generated for each assay. Table 3 Summary of Correlation Studies BN ProSpec System versus BNäII System Sample Number (n) Coefficient of Correlation (r) Regression 1 N Antiserum to Human IgG/Serum y = 00x N Antiserum to Human IgA/Serum y = 60x N Antiserum to Human IgM/Serum y = 27x N Antiserum to Human C3c y = 96x N Antiserum to Human C y = 43x N Antiserum to Human Transferrin/Serum 7 N Antiserum to Human Albumin/Serum 8 N Antiserum to Human α1- Antitrypsin 9 N Antiserum to Human α2- Macroglobulin 10 N Antiserum to Human Haptoglobin y = 10x y = 51x y = 35x y = 53x y = 11x N Antiserum to Human α1-acid y = 35x Glycoprotein 12 N Antiserum to Human Prealbumin y = 49x N Latex stfr y = 00x N Antiserum to Human Ceruloplasmin 16 N Antiserum to Human Retinol binding Protein 17 N Antiserum to Human Ig/L-chain, kappa 18 N Antiserum to Human Ig/L-chain, lambda y = 49x y = 75x y = 74x y = 73x

7 Table 3 (Cont.) Summary of Correlation Studies BN ProSpec System versus BNäII System Sample Number (n) Coefficient of Correlation (r) Regression 19 Total Protein/Serum y = 69x N Antiserum to Human y = 24x 0.09 Transferrin/Urine 29 N Antiserum to Human α y = 93x Microglobulin/Urine 30 N Antiserum to Human y = 70x Apolipoprotein A-I 31 N Antiserum to Human y = 87x Apolipoprotein B 40 N Antiserum to Human Fibrinogen y = 45x N Antiserum to Human y = 44x Antithrombin III 42 N High Sensitivity CRP y = 16x N Antiserum to Human y = 55x Plasminogen 50 N Latex Ferritin y = 69x N Latex RF y = 34x N Latex ASL y = 23x N Latex IgE y = 29x 1 58 N Latex ADNase B y = 30x N Latex Myoglobin y = 46x N Antiserum to Human IgG/CSF y = 98x N Latex IgA/CSF y = 53x N Antiserum to Human C1 Inhibitor Serum/ Plasma y = 34x y = 40x N High Sensitivity CRP y = 63x In addition to correlation studies conducted on the BN ProSpecä System versus the BNäII System, the sixteen external sites evaluated selected analytes on the BN ProSpecä System versus the BNäA System or the BNä100 System. 7

8 Selected analytes were also tested on several other manufacturer s systems. These include the Beckman Array, the Beckman Immage, and the Roche COBAS Integra. Table 4 Summary of Additional Correlation Studies BN ProSpec System versus BN 100 System Sample Number (n) Coefficient of Correlation (r) Regression 6 N Antiserum to Human y = 47x Transferrin/Serum 10 N Antiserum to Human y = 98x 0.03 Haptoglobin 30 N Antiserum to Human y = 13x Apolipoprotein A-I 31 N Antiserum to Human y = 08x Apolipoprotein B 50 N Latex Ferritin y = 80x N High Sensitivity CRP y = 53x 0.57 Table 5 Summary of Additional Correlation Studies BN ProSpec System versus BNäA System Sample Number (n) Coefficient of Correlation (r) Regression 6 N Antiserum to Human Transferrin/Serum 12 N Antiserum to Human Prealbumin 17 N Antiserum to Human Ig/Lchain, kappa 18 N Antiserum to Human Ig/Lchain, lambda 70 N Antiserum to Human C1 Inhibitor/Serum y = 79x y = 21x y = 10x y = 65x y = 00x

9 Table 6 Summary of Additional Correlation Studies BN ProSpec System versus Beckman Immage Sample Number (n) Coefficient of Correlation (r) Regression 1 N Antiserum to Human IgG /Serum 2 N Antiserum to Human IgA /Serum 3 N Antiserum to Human IgM /Serum y = 45x y = 59x y = 78x N Antiserum to Human C3c y = 49x N Antiserum to Human C y = 61x N Antiserum to Human Ig/Lchain, kappa/urine y = 0.309x N Antiserum to Human IgG y = 85x /Urine 27 N Antiserum to Human y = 09x 9 Albumin/Urine 55 N Latex ASL y = 93x N Antiserum to Human IgG y = 49x + 6 /CSF 62 N Latex IgA /CSF y = 56x N Latex IgM /CSF y = 94x N Antiserum to Human y = 07x Albumin /CSF 71 N High Sensitivity CRP y = 28x 3 9

10 Table 7 Summary of Additional Correlation Studies BN ProSpec System versus Beckman Array Sample Number (n) Coefficient of Correlation (r) Regression 6 N Antiserum to Human Transferrin/Serum 12 N Antiserum to Human Prealbumin 30 N Antiserum to Human Apolipoprotein A-I 31 N Antiserum to Human Apolipoprotein B y = 29x y = 07x y = 86x y = 69x Table 8 Summary of Additional Correlation Studies BN ProSpec System versus Roche COBAS Integra Sample Number (n) Coefficient of Correlation (r) Regression 1 N Antiserum to Human IgG y = 09x N Antiserum to Human IgA y = 93x N Antiserum to Human IgM y = 0.691x N Antiserum to Human Albumin y = 43x N High Sensitivity CRP y = 08x N Latex ASL y = 15x

11 Interference Testing Interference testing was performed for the assays listed in Table 9 to determine the effect of lipemia, hemolysis, icterus, and rheumatoid factors on plasma protein assays when analyzed on the BN ProSpecä System. rmal sample preparations (serum or plasma) were spiked with increasing concentrations of triglycerides ( 1924 mg/dl), free hemoglobin ( 1000 mg/dl), total bilirubin ( 60 mg/dl), and rheumatoid factor (up to 3080 IU/mL). The concentrations listed in Table 9 reflect the levels of endogenous substances at which interference was first noted for the specific assay being tested. Table 9 BN ProSpec System Summary of Interference Studies Endogenous Substances Triglycerides (mg/dl) Hemoglobin (mg/dl) Bilirubin (mg/dl) RF (IU/mL) 1 N Antiserum to Human IgG N/A 2 N Antiserum to Human IgA N/A 3 N Antiserum to Human IgM ** N/A 4 N Antiserum to Human C3c N/A 5 N Antiserum to Human C N/A 6 N Antiserum to Human Transferrin/Serum N/A 7 N Antiserum to Human Albumin/Serum N/A 8 N Antiserum to Human α1-antitrypsin N/A 9 N Antiserum to Human α2-macroglobulin N/A 10 N Antiserum to Human Haptoglobin N/A 11 N Antiserum to Human α1-acid Glycoprotein N/A 12 N Antiserum to Human Prealbumin N/A 13 N Latex stfr N/A 15 N Antiserum to Human Ceruloplasmin N/A 16 N Antiserum to Human Retinol binding Protein N/A 17 N Antiserum to Human Ig/L-chain, kappa N/A 18 N Antiserum to Human Ig/L-chain, lambda N/A 26 N Antiserum to Human Transferrin/Urine N/A ** Interference noted at all levels of triglycerides tested in the study (243 mg/dl to 1824 mg/dl). 11

12 Table 9 (Cont.) BN ProSpec System Summary of Interference Studies Endogenous Substances Triglycerides (mg/dl) Hemoglobin (mg/dl) Bilirubin (mg/dl) RF (IU/mL) 30 N Antiserum to Human Apolipoprotein A-I N/A 31 N Antiserum to Human Apolipoprotein B N/A 40 N Antiserum to Human Fibrinogen N/A 41 N Antiserum to Human Antithrombin III ** N/A 42 N High Sensitivity CRP N/A 43 N Antiserum to Human Plasminogen ** N/A 50 N Latex Ferritin N Latex RF N/A 55 N Latex ASL N/A 57 N Latex IgE N Latex ADNase B ** N Latex Myoglobin N Latex IgA N/A 70 N Antiserum to Human C1 Inhibitor/ Serum Plasma N/A N/A 71 N High Sensitivity CRP N/A ** Interference noted at all levels of triglycerides tested in the study (243 mg/dl to 1824 mg/dl). 12

13 Linearity Study Samples with a high concentration of analyte were serially diluted down to the lower end of the measuring range. Each dilution was tested in replicates of five, and the mean % recovery was determined [% Recovery = (measured concentration / theoretical concentration) x 100]. Linear range is presented in Table 10 for those assays tested. Table 10 BN ProSpec System Summary of Linearity Studies Unit Linear range 1 N Antiserum to Human IgG mg/dl N Antiserum to Human IgA mg/dl N Antiserum to Human IgM mg/dl N Antiserum to Human C3c mg/dl N Antiserum to Human C4 mg/dl N Antiserum to Human mg/dl Transferrin/Serum 7 N Antiserum to Human Albumin mg/dl N Antiserum to Human α1-antitrypsin mg/dl N Antiserum to Human α2- mg/dl Macroglobulin 10 N Antiserum to Human Haptoglobin mg/dl N Antiserum to Human α1-acid mg/dl Glycoprotein 12 N Antiserum to Human Prealbumin mg/dl N Latex stfr mg/l N Antiserum to Human Ceruloplasmin mg/dl N Antiserum to Human Retinol binding Protein 17 N Antiserum to Human Ig/L-chain, kappa 18 N antiserum to Human Ig/L-chain, lambda 26 N Antiserum to Human Transferrin/Urine 29 N Antiserum to Human α1- Microglobulin/Urine mg/dl mg/dl mg/dl mg/l mg/l

14 Table 10(Cont.) BN ProSpec System Summary of Linearity Studies Unit Linear range 30 N Antiserum to Human Apolipoprotein A-I mg/dl N Antiserum to Human Apolipoprotein B mg/dl N Antiserum to Human Fibrinogen mg/dl N Antiserum to Human Antithrombin III mg/dl N High Sensitivity CRP mg/l N Antiserum to Human Plasminogen mg/dl N Latex Ferritin µg/l N Latex RF IU/mL N Latex ASL IU/mL N Latex IgE IU/mL N Latex ADNase B U/mL N Latex Myoglobin µg/l N Antiserum to Human IgG/CSF mg/l N Latex IgA/CSF mg/l N Latex IgA/Serum mg/l N Antiserum to Human C1 Inhibitor mg/dl N High Sensitivity CRP mg/l Conclusions The BN ProSpecä System demonstrates at least eight (8) weeks calibration stability for all assays tested. The BN ProSpecä System has demonstrated excellent precision. The BN ProSpecä System displays very good correlation to the BNäII, the BNäA, and the BNä100 Systems. Selected analytes tested on the BN ProSpecä System demonstrate good correlation to other manufacturer s systems with few exceptions. s tested on the BN ProSpecä System exhibit minimal, if any, interference from elevated levels of lipemia, icterus, hemolysis, and rheumatoid factors. Testing on the BN ProSpecä System confirms linearity across the assay range of all analytes tested. BN ProSpecä System,BNäII, BNä 100, BNäA are trademarks of Dade Behring Marburg GmbH. 14