Transcript Document 7655292

Evaluation data on the new
Roche cobas s 201 system with
the cobas TaqScreen MPX test
Lutz Lutz
PichlPichl,
, DRK-Blutspendedienst
Volkmar SchottstedtWest
German
Red
Cross
Blood Transfusion Service
Christine
Jork,
DRK-Blutspendedienst
NSTOBWest,
Hagen, Germany
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~ 4,000 - 6,000 samples/ day
Hagen
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cobas s 201 system
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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 HBV
by multiplex PCR in a large blood bank setting
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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
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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)
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Reproducibility
 3 Run Controls:
HBV: 30 IU/ml
HCV: 50 IU/ml
HIV-1: 160 IU/ml
 8 replicates of each control were analysed on 4 days
by 2 operators with 2 different kit lots
HBV
HCV
HIV-1
Neg
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Reactive results/ tested replicates
Day 1
Day 2
Day 3
Day 4
8/8
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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.
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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
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Screening minipools (n =96) from
blood donations
Minipools
tested
365
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Non
reactive
360
Initially
reactive
5
Invalid by
Software
0
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