DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

Experience with Roche´s COBAS AmpliPrep/COBAS TaqMan96 automated PCR

system in two large blood bank settings

Lutz Pichl , DRK-Blutspendedienst WestChristine Jork, DRK-Blutspendedienst NSTOBChristine Jork, DRK-Blutspendedienst NSTOB

Lutz Pichl , DRK-Blutspendedienst West

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

DRK-BSD West

DRK-BSD NSTOB

Springe

Hagen

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

BSD NSTOB BSD WestDonations/year (2005)

Samples/Day1.15 mio~ 4.000

1.26 mio~ 4.000

Viral parameters HCV, HIV-1, HBV, HAV, PVB19

Routine testing with CAP/CTM96 since 1st Nov. 2005 1st July 2005

CAP/CTM96 Systems 6 x 9 x

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

BSD NSTOB BSD WestCAP/CTM96 combined via docking station for HBV, HCV and HIV-1

CAP/CTM96 combined via docking station for HBV, HCV and HIV-1+ ventilation shaft for waste air

2 CAP/CTM96 for HCV

2 CAP/CTM96 for HIV-1

1 CAP/CTM96 for HBV

1 CAP for TNAI HAV/PVB19 RealArt LC PCR Reagents (QIAGEN Diagnostics), LightCycler 1.2 (Roche)

3 CAP/CTM96 for HCV

3 CAP/CTM96 for HIV-1

3 CAP/CTM96 for HBV

2 CAP for TNAI HAV/PVB19 RealArt LC PCR Reagents (QIAGEN Diagnostics), LightCycler 2.0 (Roche)

System configuration

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

BSD NSTOB BSD West

Samples tested 668,469 (11/2005 – 05/2006)

Pool size 96 96

Invalidity rate (HCV / HIV-1 / HBV)

Failure rate0.9 %

(hardware error)< 0.2 %

(clot, hardware error)

NAT only positives

842,751(07/2005 -05/2006)

HCV: 2 (05) 2 (06)HBV: 1

HCV: Ø 0.27 %HIV-1: Ø 0.12 %HBV: Ø 0.05 %

HIV-1: 2 (06)HBV: 1 (06)

HCV: Ø 1.68 %HIV-1: Ø 0.15 %HBV: Ø 0.14 %

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

95% detection limits BSD NSTOB BSD West

HBV 675 IUWHO/ml SD 251 IUWHO/ml SD

HCV 582 IUWHO/ml SD 624 IUWHO/ml SD

HIV-1 4315 IUWHO/ml SD 7876 IUWHO/ml SD

PVB19 5784 IUWHO/ml SD 5735 IUWHO/ml SD

HAV 21420 IUWHO/ml SD 6157 IUWHO/ml SD

SD = single donation in pool of 96 samples

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

Turn around time

CAP/CTM M

ainten

ance

Sample

Prep

aratio

n

1 h

NA Extr

actio

n

2 h

Amplific

ation

&

Detecti

on

HIV-1 3h 15‘

HCV 2h 40‘

HBV 2h 10‘

HAV/PVB 50‘

Poolin

g

0,5 h

Result

Vali

datio

n

Data Tran

sfer (

LIMS)

15-30‘

6 a.m. ~1 p.m.

DRK-BSD NSTOB & West 19th SoGAT Meeting, 14/15 June 2006, Bern

Pro Con

fully automatedintensive maintenance caused by

high system complexity

short hands-on timeadditional manual result

interpretation

IVD conform only for single samplesoff label use for minipool testing;

extensive validation studies

virus enrichment steps unnecessary high sample volume (1050 µl brutto volume)

automation of front end sample handling is users responsibility

in-house solutions

Pros & Cons of CAP/CTM96 System