Abstract
Background:Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies. Methods:The purpose of this study was to extract the maximum amount of gDNA from DBS on Whatman 903 protein saver card. gDNA was extracted through column (Qiagen) & magnetic bead based (Invitrogen) methods. Quantification of extracted gDNA was performed with a spectrophotometer, fluorometer, and integrity analyzed by agarose gel electrophoresis. Result:Large variation was observed in quantity & purity (260/280 ratio, 1.8-2.9) of the extracted gDNA. The intact gDNA bands on the electrophoresis gel reflect the robustness of DBS for gDNA even after prolonged storage time. The extracted gDNA amount 2.16 – 24 ng/µl is sufficient for its PCR based downstream application, but unfortunately it can’t be used for whole genome sequencing or genotyping from extracted gDNA. Sequencing or genotyping can be achieved by after increasing template copy number through whole genome amplification of extracted gDNA. The obtained results create a base for future research to develop high-throughput research and extraction methods from blood samples.Conclusion:The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies.
Highlights
The concept of using dried blood spot (DBS) in new born screening was presented by Guthrie and Susie in 19631
Today Dried blood spot (DBS) samples are utilized to test for a variety of health related markers including; infectious pathogens, HbA1c, total cholesterol, creatinine, uric acid, low density lipoprotein (LDL), high density lipoprotein (HDL), very low density lipoprotein (VLDL), Triglyceride and many more2–4
DBS samples were collected at health checkup camps in rural and urban areas of Bangalore city through a sample registration system (SRS). 3000 DBS samples were prepared through finger prick method by using lancet (Accu Chek Softclix Lancet, Roche), puncture the finger site using lancet, drop of blood form which is lightly touch the circle of filter paper cards (GE Health Care Life Science, Catalog no. 10534612) and form valid DBS during health checkup camp by Centre for Global Health Research (CGHR) at Bangalore unit and transported to Tata memorial Centre (TMC) Mumbai for further analysis
Summary
The concept of using dried blood spot (DBS) in new born screening was presented by Guthrie and Susie in 19631. In epidemiological research the protein saver cards act as information storage devices in terms of blood based analytes and provide genetic, environmental, immunological information. Genomic DNA (gDNA) is a very robust and stable biological sample when stored on paper cards, and has been used for many decades. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies. Conclusion: The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies
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