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Performance evaluation of the QIAGEN EZ1 DSP virus kit with Abbott RealTime HIV-1, HCV and HBV assays

Abstract number: 1733_1230

Schneider G., Kuper K., Abravaya K., Mullen C., Schmidt M., Sprenger-Haussels M.

Objectives: Automated systems must meet the demands of routine diagnostics laboratories with regard to performance characteristics and compatibility with downstream assays. The EZ1 DSP Virus system is a generic sample preparation system utilising QIAGEN's second generation magnetic-bead technology on the QIAGEN BioRobot EZ1 DSP workstation for isolation and purification of viral nucleic acids from human plasma or serum. The improved EZ1 DSP Virus protocol is taking advantage of optimised lysis conditions as well as increased binding and washing efficiencies. The performance of the QIAGEN EZ1 DSP Virus Kit was evaluated in combination with the Abbott RealTime HIV-1, HCV and HBV assays.

Methods: RNA viruses (HIV-1 and HCV) and a DNA virus (HBV) were spiked into negative human plasma or serum and processed in batches of six. An internal control was added to each sample to control for recovery and inhibition. Nucleic acids were extracted from 0.4 ml of plasma or serum using the EZ1 DSP Virus Kit and were eluted in a volume of 90 microliters. The resulting nucleic acids were analysed by the appropriate RealTime assay on the Abbott m2000rt thermalcycler/reader according to the package inserts. Run controls were included on every thermalcycler run.

Results: RealTime assay results were linear over the ranges tested. The observed inter-assay SDs were less than or equal to 0.13 log except at the lowest concentration tested in each assay. The EZ1 DSP Virus kit enabled detection of low levels of virus. Good correlation between the BioRobot EZ1 and the Abbott m2000sp was observed. EZ1 DSP Virus kit reagent lot-to-lot variability was evaluated. No detectable false positives among negative samples were detected in six carryover challenge runs. No inhibition was observed in the presence of potentially interfering substances.

Conclusions: The QIAGEN EZ1 DSP Virus system enables reproducible purification of viral nucleic acids from human plasma or serum and allows for reliable, sensitive and specific detection and quantification of RNA and DNA viruses by the Abbott RealTime HIV-1, HCV and HBV assays.

*Under development for in-vitro-diagnostic use in Europe. The EZ1 DSP Virus Kit as described in this poster will not be available in the USA.

** Abbott RealTime HIV-1 and HCV are CE marked, not available in the USA. Abbott RealTime HBV is in development. Abbott Molecular makes no Abbott RealTime assay performance claims using the EZ1 DSP Virus Kit.

Session Details

Date: 31/03/2007
Time: 00:00-00:00
Session name: European Society of Clinical Microbiology and Infectious Diseases
Subject:
Location: ICC, Munich, Germany
Presentation type:
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