Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Miklau, F.
Comparison of automated extraction platforms on quantification of plasma hepatitis D virus RNA.
Humanmedizin; [ Diplomarbeit ] Graz Medical University; 2020. pp. 33
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Kessler Harald
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- Abstract:
- Background: Hepatitis D (HDV) is the most severe and rapidly progressive form of chronic viral hepatitis needing a reliable diagnostic approach. The aim of this study was to make results comparable when using the CE/IVD-labeled RoboGene® HDV RNA Quantification Kit 2.0 (Analytik Jena AG) with two different laboratory settings. Materials and Methods: All samples were extracted either with the eMAG® (bioMérieux S.A., Marcy l´Etoile, France) or the MagNA Pure 24 (Roche Molecular Diagnostics, Rotkreuz, Switzerland) platform. Amplification and detection were performed on the LightCycler 480II instrument (Roche). After determination of correction factors (CF) utilizing the 1st WHO International Standard for HDV RNA, the limit of detection (LOD) was determined for both test systems followed by accuracy testing using a reference panel. Furthermore, 30 clinical plasma samples (25 of them HDV RNA positive and 5 of them HDV RNA negative) were tested and results compared. Results: The CF was 922 for the test system including the eMAG® and 190 for the test system including the MagNA Pure 24. LODs were found to be 2.65 log10 IU/mL and 2.37 log10 IU/mL, respectively. One member of the reference panel containing 2.03 log10 IU/mL tested negative with both test systems and another member containing 3.11 log10 IU/mL gave a negative result with the test system including the eMAG®. Of 25 HDV RNA positive samples, 15 tested positive with both test systems, 2 gave discrepant results, and in 8 samples, HDV RNA was not detected with both of the methods. All clinical samples that did not contain HDV RNA tested negative with both test systems. Conclusions: Quantification of plasma HDV RNA depends on the extraction platform used. To make results comparable, it is mandatory to determine the specific CF for the test system employed. Harmonization of results is essential for monitoring patients undergoing anti-HDV therapy and making results obtained from clinical studies comparable.