GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 1
Evaluation data on the new Roche cobas s 201 system with the cobas TaqScreen MPX test
Lutz Pichl , DRK-Blutspendedienst West
Christine Jork, DRK-Blutspendedienst NSTOB
Lutz Pichl, Volkmar SchottstedtGerman Red Cross Blood Transfusion Service West,
Hagen, Germany
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 2
Hagen
~ 4,000 - 6,000 samples/ day
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 3
cobas s 201 system
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 4
Aim of the study
evaluation of the Roche cobas s 201 System in combination with the TaqScreen MPX test
feasibility to screen plasma minipools(pools size 96 samples) for HCV, HIV-1,HIV-1 O, HIV-2 and HBVby multiplex PCR in a large blood bank setting
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 5
Conditions
Fully automated, IVD-conf. , CE-marked system Focus on the performance of the MPX test
rather than on hardware features of the s 201(lab sample pooling unit and PCR unit with CAP/CTM systems have been operated since 2005 under routine screening conditions)
minipools are processed by the integrated Hamilton Microlab Star pipettor like single units
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 6
Cross-Contamination Study
16 high titre, positive donor samples (5 HBV-DNA pos., 5 HCV-DNA pos., 5 HIV-1 pos.) and 16 non-reactive donor samples in alternating manner of 32 position on 2 racks
titres HBV: 8.0 EE8, HCV: 1.0 EE7, HIV-1: 1.0 EE6 2 different loading schemes were analysed result: no cross-contamination observed except
1 false (weak-) positive HCV (Ct 46.3 vs. Ct 18.4)
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 7
Reproducibility
Reactive results/ tested replicatesDay 1 Day 2 Day 3 Day 4
HBV 8/8 8/8 8/8 8/8HCV 8/8 8/8 8/8 8/8HIV-1 8/8 8/8 8/8 8/8Neg 8/8 8/8 8/8 8/8
3 Run Controls: HBV: 30 IU/mlHCV: 50 IU/mlHIV-1: 160 IU/ml
8 replicates of each control were analysed on 4 daysby 2 operators with 2 different kit lots
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 8
Potential interference with endogenous substances
Haemolysed plasma samples consisted of up to 20% (v/v) lysed red cell concentrate (12,550 mg/dl Hb)
lipaemic plasma samples (871 mg/dl triglycerides) “green colored” plasma from female donors under hormonal
treatment Individual plasma samples were seperately spiked with HBV (30
IU/ml), HIV-1M (50 IU/ml) and HCV (160 IU/ml),final concentrations
5 (haemolytic) to 20 replicates (lipaemic/ green) were analysed Induced high concentrations of hemoglobin, fat and other
possibly interfering substances showed no inhibitory effect on PCR results at definite concentrations.
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 9
Genotype Inclusivity
• NIBSC HCV RNA genotype panel 02/202 for NAT,final conc. 100 IU/ml all panel members detected
• Acrometrix VQC PeliCheck HBV-DNA genotype panel (1,000 cop/ml) all panel members detected
• NIBSC 1st Int. Ref. Panel 01/466 for HIV-1 RNA all panel members detected except for HIV 01-466 (group O) further investigation
GRC-BTS West 20th SoGAT Meeting, 12 - 13 June 2007, Warsaw 10
Screening minipools (n =96) from blood donations
Minipoolstested
Nonreactive
Initiallyreactive
Invalid bySoftware
365 360 5 0