Optimising stool preservation methods to overcome cold chain constraints for the diagnosis of enteric RNA viral diseases

Roop Dhillon (University of Birmingham, UK)

12:39 - 12:51 Wednesday 15 April Morning

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Abstract

Traveller’s diarrhoea is an enteric dysfunction caused by long distance international travel. Bacterial causes predominate, but the RNA viruses rotavirus and norovirus cause up to 16% of cases annually. Accurate detection of RNA viruses remains a challenge when cold-chain storage is limited since RNA degradation compromises detection and diagnostics leading to false negatives and a loss in surveillance. Here we evaluated three commercial stool storage solutions under conditions relevant to field diagnostics - DNA/RNA Shield (Zymo Research), OmniGene GUT OM-200 and OMR-205 (DNA Genotek). Two RNA phages, the enveloped Pseudomonas savastanoi phage Phi6 and the non-enveloped Escherichia coli phage MS2 were used as surrogates for human RNA viruses. Phage lysates were tested alone or co-incubated with ZymoBIOTICS fecal reference material, stored for 1,3 or 7 days at -20⁰C, 4⁰C, or room temperature. Genome copy number was determined by RT-qPCR (cT < 35) and was used to quantify diagnostic viability. DNA/RNA Shield outperformed both OM-200 and OMR-205, maintaining > 90% genome recovery rates at both -20⁰C and 4⁰C. Diagnostically relevant levels of genome copy number was maintained over 7 days. OMR-205 could not preserve viral RNA for any period of time. These findings highlight the suitability of DNA/RNA Shield for the preservation of RNA viruses in stool samples taken from low-resource settings. This enables diagnosis for enteric RNA viruses without access to ultra-low freezing or advanced diagnostic tools.

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