Evaluation of the AcroMetrix® OptiChallenge Inhibition Panel in different QIAGEN® sample preparation protocols automated on the QIAcube
Abstract number: P774
Kupfer C., Lee Y., Schulte G., Sprenger-Haussels M.
Objectives: The Clinical and Laboratory Standards Institute recommends that all clinical assay performances should be evaluated in the presence of potentially interfering substances to validate assays, to quantify interference effects, and to confirm interference in patient samples (CLSI EP-7-A2). This is particularly important for homebrew assays, where no data is available from the manufacturer on the performance of the assay in the presence of such substances. Clinical laboratories are responsible for verifying in all assays that the levels of interfering substances in their patient population are within an acceptable range.
Methods: The AcroMetrix OptiChallenge Inhibition Panel is a 7-member panel that mimics patient plasma containing potentially interfering substances that frequently occur due to clinical conditions such as hemolysis, icterus, or lipaemia.
We have evaluated the interference effect of the inhibition panel members on the performance of different silica-based sample preparation protocols on the QIAcube and their manual reference methods. Common bloodborne viruses, such as HCV and HBV, and different internal controls were spiked into each panel member in equal quantities.
Results: All tested automated sample-preparation protocols tested performed well and removed a good amount of the interfering substances when specific amounts of each analyte spiked were quantified in the presence of potentially interfering substances and compared with no-interference control. Results of the automated protocols were comparable to the manual reference methods. Different extraction and amplification chemistries show different susceptibilities to single interfering substances.
Conclusion: The automated QIAGEN extraction protocols, tested for isolation of viral RNA and DNA, have been proven to serve as reliable and robust methods. The inhibition panel was only minimally diluted, so the extraction systems were tested at the highest possible level of interference. The tested protocols were able to remove the majority of potentially interfering substances.
The OptiChallenge Inhibition Panel provides an effective tool to monitor the performance of nucleic acid extraction and amplification methods and simplifies the required validation processes by providing plasma samples that mimic interfering patient samples. This panel could be useful in QC release testing of DNA and RNA extraction and amplification systems.
|Session name:||18th European Congress of Clinical Microbiology and Infectious Diseases|
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