Iran J Parasitol: Vol. 14, No. 1, Jan-Mar 2019, pp.29-40 29 Available at: http://ijpa.tums.ac.ir Original Article A Simple and Cost-Effective Freeze-Thaw Based Method for Plasmodium DNA Extraction from Dried Blood Spot Supriya SHARMA, Riti MANN, Sandeep KUMAR, Neelima MISHRA, Bina SRIVASTAVA, Neena VALECHA, *Anupkumar R. ANVIKAR ICMR-National Institute of Malaria Research, Sector 8, Dwarka, New Delhi, 110077, India Received 15 Jan 2018 Accepted 21 May 2018 Abstract Background: Available DNA isolation methods for Plasmodium involve numerous processing steps, adding to the cost and conferring risk of contamination. Here we devise a simple and cost-effective method for direct extraction of Plasmodium DNA from dried filter paper spot (DBS), appropriate for resource-limited setups. Methods: The protocol involves simple freezing and thawing of DBS, neither involves any purification step nor any chemical reagent. The method was assessed in terms of DNA quantity, PCR detection sensitivity, time requirement, cost effec- tiveness, labor intensiveness and degree of shearing. The reliability of this method was confirmed by comparing it with other in use methods for Plasmodium DNA isolation. Results: Pure DNA was obtained with this method, as exemplified by the absorb- ance ratio (260nm /280nm) of 1.2. The protocol produced digestible, PCR-grade genomic DNA, also found to be suitable for sequencing. DNA isolated remained stable and retained its integrity after storage for one month at 4 0 C. Conclusion: Our process substantiated as efficient, reproducible, simple, fast, and inexpensive. Development of this optimized freeze-thaw based DNA extraction method for malaria parasite may provide a valuable tool for molecular analysis in resource-limited setups. This is the first report of DNA extraction from DBS of Plasmodium utilizing freeze-thaw. Keywords: Plasmodium; DNA isolation; Freezing; Thawing; PCR * Correspondence Email: anvikar@gmail.com Introduction alaria caused by Plasmodium sp. is a serious global health problem, lead- ing to about 584,000 fatalities an- nually (1). Apt and correct diagnosis is neces- sary for prevention, control and suitable treatment of malaria. Accurate diagnosis can- M Iranian Society of Parasitology http://isp.tums.ac.ir Iran J Parasitol Open access Journal at http://ijpa.tums.ac.ir Tehran University of Medical Sciences Publication http://tums.ac.ir