Cost-Effective and Scalable DNA Extraction Method from Dried Blood Spots

Abstract

BACKGROUND: Dried blood spot (DBS) samples have been widely used in newborn screening (NBS) for the early identification of disease to facilitate the presymptomatic treatment of congenital diseases in newborns. As molecular genetics knowledge and technology progresses, there is an increased demand on NBS programs for molecular testing and a need to establish reliable, low-cost methods to perform those analyses. Here we report a flexible, cost-efficient, high-throughput DNA extraction method from DBS adaptable to small-and large-scale screening settings.

Development of DNA Confirmatory and High-Risk Diagnostic Testing for Newborns Using Targeted Next-Generation DNA Sequencing

“…26,37 The cost of NBDx (Supplementary Note S2 online) is significantly less than that of WES, and both tests are expected to be similar in price range to diagnostic tests currently on the market and therefore should enable replacement of single-gene tests, as justified by delays and increased patient-management costs. 6,10,11 We established performance benchmarks supporting direct clinical use similar to WGS newborn/pediatric testing of Mendelian diseases.…”

“…Recently, second-tier testing using amplicon NGS has been proposed for severe combined immunodeficiency disease (OMIM 300400) after primary screening of T-cell receptor excision circles from DBS. 25,26 We undertook the development of fast-turnaround, minimally invasive, and cost-effective clinical sequencing and reporting in newborns, allowing a simplified testing menu that utilizes both WES and a more focused panel approach (NBDx), along with in silico gene filters (Figure 1a and Table 1). In a retrospective proof-of-concept study, we determined the performance in the context of sequence variants associated with metabolic and other genetic disorders responsible for common phenotypes in the neonate.…”

Superparamagnetic-bead Based Method: An Effective DNA Extraction from Dried Blood Spots (DBS) for Diagnostic PCR

“…The efficiency of DNA recovered from DBS in PCR-based techniques was an issue of interest of different researchers, where the extraction method was considered as valid and reliable not only based on the yield but also on the quality of the DNA to perform the progressive molecular diagnostic techniques [15].…”

“…The success of these DBS as an easy and cost-effective method for collecting, archiving, and transporting blood specimens has inspired researchers to explore better DNA extraction and PCR-based molecular testing methods for these samples [9][10][11][12]. The DBS sampling is an effective alternative for lymphocyte pellets because DBS sampling requires only a few droplets of whole blood, not necessarily venous blood, and can be directly spotted on Guthrie cards or filter papers [13][14][15].…”

“…The success of these DBS as an easy and cost-effective method for collecting, archiving, and transporting blood specimens has inspired researchers to explore better DNA extraction and PCR-based molecular testing methods for these samples [9][10][11][12]. The DBS sampling is an effective alternative for lymphocyte pellets because DBS sampling requires only a few droplets of whole blood, not necessarily venous blood, and can be directly spotted on Guthrie cards or filter papers [13][14][15].The commonly used methods and techniques for extracting genomic DNA from DBS are varied in terms of extraction principle, reagents used, and quantity and quality of the extracted DNA. Manual protocols include the extraction of DNA with Tris-EDTA buffer, methanol, and Chelex-100 [16][17][18][19].…”