Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Radler, A.
Evaluation of a novel assay for detection of norovirus RNA based on a rapid amplification and detection protocol using two different extraction platforms.
Humanmedizin; [ Diplomarbeit ] Graz Medical University ; 2020. pp. 34
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Kessler Harald
- Stelzl Evelyn
- Altmetrics:
- Abstract:
- Background: Noroviruses are significant pathogens causing infectious gastroenteritis. It is of utmost importance that patients infected with norovirus are identified as early as possible. Objectives: The aim of this study was to compare results obtained by the Anchor Norovirus PCR Kit (Anchor Diagnostics) with the RIDA®GENE Norovirus assay (R-Biopharm). Materials and Methods: The analytical performance of the Anchor Norovirus PCR Kit was assessed utilizing two international pilot study panels. The clinical performance was evaluated with 109 anonymized residual clinical stool samples. For detection of norovirus RNA, all samples were extracted on the EMAG® (bioMerieux) platform and the MagNA Pure 24 (Roche) instrument followed by amplification and detection using the Anchor Norovirus PCR Kit. The RIDA®GENE Norovirus assay in combination with the EMAG® platform served as reference test system. Amplification and detection were performed on the Light Cycler® 480 II (Roche) for all 3 test systems. Results: When the analytical performance was assessed, all positive and negative panel members were identified as positive and negative, respectively. When the clinical performance was evaluated, 69 samples tested positive and 40 negative with the reference test system. When the Anchor Norovirus PCR Kit was used in combination with the EMAG® platform, all 69 positives tested positive, 36 of 40 negatives tested negative, and 4 samples showed inhibition; in combination with the MagNA Pure 24 platform, all 69 positives tested positive, 4 of 40 negatives tested negative, and 36 samples showed inhibition. All valid results were found to be concordant. The overall time required for an extraction of 8 samples on the EMAG® platform was 80 minutes, the hands-on time 20 minutes. Corresponding times when using the MagNA Pure 24 platform were 40 and 15 minutes, respectively. Conclusions: Rapid detection of norovirus RNA in stool samples is advisable; however, the Anchor Norovirus PCR Kit in combination with the MagNA Pure 24 platform showed minor performance due to the significantly increased number of inhibitions. In combination with the EMAG® platform, significantly less inhibitions were observed. The Anchor Norovirus PCR Kit in combination with the MagNA Pure 24 platform showed a significantly shorter turn-around time; however, only 8 samples can be run in parallel.