An assessment of three noncommercial DNA extraction methods from dried blood spots for beta‐thalassaemia mutation identification

Abstract

The NaOH method is a simple method that uses minimal equipment and reagents that make it labour- and cost-effective. This method could be adopted by poorer countries to extract DNA for beta-thalassaemia mutation characterization.

The use of Taqman genotyping assays for rapid confirmation of β‐thalassaemia mutations in the Malays: accurate diagnosis with low DNA concentrations

“…Collection of dried blood spots on filter paper blotters (Guthrie spots) have been used for neonatal screening as it is convenient for sample collection, handling and shipment. One of the concerns using Guthrie spots in neonatal screening is the low yield of DNA, and many researchers have attempted various techniques to enhance the yield such as microextraction, direct amplification of DNA using methanol, sodium hydroxide (NaOH), sodium hyrdoxide (NaCl) or sodium dodecyl sulphate (SDS) method . Prenatal diagnosis is carried out for thalassaemia using invasive procedures using amniocentesis or chronic villous sampling (CVS) techniques and noninvasive procedure by analysis of circulatory cell‐free foetal DNA (cffDNA) sequences in maternal plasma.…”

Current and future use of “dried blood spot” analyses in clinical chemistry

“…DNA micro-extraction from dried blood has allowed the detection of mutations responsible for diseases such as cystic fibrosis [124], X fragile syndrome [107], spinal muscular atrophy [106], cancers [77] and thalassemia [105]. DBS, which is a fairly inexpensive sampling and storage method, is also a good choice for genetic material biobanks [125].…”

Standardization of an organic DNA extraction method from dried blood spots and its downstream molecular applications for neonatal screening and diagnostic confirmation of lysosomal disorders

“…DBS sampling has since emerged as an appropriate method for the collection, transportation, analysis, and storage of biological fluids due to its simplicity, reduced invasiveness, and low cost [2][3][4] . It is also the best option when venipuncture, transportation, and storage conditions are not favorable 5 , which is why DBS are still used as samples for newborn screening (NBS) 6 .…”

“…The use of DBS for diagnosis has increased throughout the years 7 and, despite some limitations (small amount of available material and risk of DNA degradation, among others), its performance in downstream molecular processes is similar or only slightly inferior to gold-standard sample types 2 . To date, DNA obtained from DBS has been used for diagnostic purposes using different molecular biology techniques, such as standard polymerase chain reaction (PCR) 5 , real-time PCR 8 , multiplex ligation-dependent probe amplification 9 , high-resolution melting analysis 10 , and different next-generation sequencing (NGS) approaches such as targeted NGS (TNGS), whole-exome sequencing, and whole-genome sequencing [11][12][13][14][15] .…”