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