RNA sequencing of archived neonatal dried blood spots

Mol Genet Metab Rep. 2016 Dec 24:10:33-37. doi: 10.1016/j.ymgmr.2016.12.004. eCollection 2017 Mar.

Abstract

Neonatal dried blood spots (DBS) are routinely collected on standard Guthrie cards for all-comprising national newborn screening programs for inborn errors of metabolism, hypothyroidism and other diseases. In Denmark, the Guthrie cards are stored at - 20 °C in the Danish Neonatal Screening Biobank and each sample is linked to elaborate social and medical registries. This provides a unique biospecimen repository to enable large population research at a perinatal level. Here, we demonstrate the feasibility to obtain gene expression data from DBS using next-generation RNA sequencing (RNA-seq). RNA-seq was performed on five males and five females. Sequencing results have an average of > 30 million reads per sample. 26,799 annotated features can be identified with 64% features detectable without fragments per kilobase of transcript per million mapped reads (FPKM) cutoff; number of detectable features dropped to 18% when FPKM ≥ 1. Sex can be discriminated using blood-based sex-specific gene set identified by the Genotype-Tissue Expression consortium. Here, we demonstrate the feasibility to acquire biologically-relevant gene expression from DBS using RNA-seq which provide a new avenue to investigate perinatal diseases in a high throughput manner.

Keywords: DBS, Dried blood spots; DNSB, Danish Neonatal Screening Biobank; Dried blood spots; GTEx, Genotype-Tissue Expression consortium; Gene expression; Guthrie Cards; Limited material; Neonatal Screening; RNA-seq.