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This study evaluated two ambient temperature storage methods, FTA cards and DESS buffer, for preserving individual and pooled infective larvae of rat parasitic nematodes (Nippostrongylus brasiliensis and Strongyloides ratti) prior to whole-genome sequencing. Results showed that both methods produced lower proportions of sequence reads mapping to reference genomes when applied to individual larvae, compared to frozen controls, with DESS buffer generally outperforming FTA cards. Importantly, when larvae were pooled in groups of 10 or 50, both ambient storage methods yielded sequencing results comparable to frozen controls.
Why it matters
These findings are relevant for field-based genomic studies of soil-transmitted helminths in endemic regions where cold-chain infrastructure is limited or unavailable, suggesting that pooled sample strategies with ambient storage could be a practical alternative to frozen storage for whole-genome sequencing efforts.
⚠️ Preprint – Noch nicht peer-reviewed
Dieser Artikel wurde noch nicht von unabhängigen Experten begutachtet. Die Ergebnisse sind vorläufig und sollten mit Vorsicht interpretiert werden.
Soil-transmitted helminth (STH) infections are a major public health burden, and there are programmes of mass drug administration that attempt to ameliorate the harm that they cause. There has been increasing use of genomics to study STH infections and other parasitic nematodes, with particular interest in whole genome sequencing (WGS). For such studies, samples are commonly stored frozen, but in settings where these infections are endemic this can be difficult, and so there would be advantages to having ambient temperature storage methods. We investigated two ambient temperature storage methods – FTA cards and DESS buffer – for infective larvae of the rat parasites Nippostrongylus brasiliensis and Strongyloides ratti, prior to DNA extraction and then WGS. Our results showed that for individual larvae stored on FTA cards or in DESS buffer, this resulted in a lower proportion of sequence reads that mapped to the reference genomes, compared to the frozen control samples. Generally, for individual larvae, DESS-storage resulted in better sequencing results than FTA-storage. However, for pools of 10 or 50 larvae, then these ambient temperature storage methods generally resulted in comparable sequence read mapping to the frozen control samples.
Source: Ambient temperature storage of individual parasitic nematode larvae for whole-genome sequencing.