The Effect of Storage and Extraction Methods on Amplification of Plasmodium falciparum DNA from Dried Blood Spots

The American Journal of Tropical Medicine and Hygiene
Alanna SchwartzBryan Greenhouse

Abstract

Extraction and amplification of DNA from dried blood spots (DBS) collected in field studies is commonly used for detection of Plasmodium falciparum. However, there have been few systematic efforts to determine the effects of storage and extraction methods on the sensitivity of DNA amplification. We investigated the effects of storage conditions, length of storage, and DNA extraction methods on amplification via three PCR-based assays using field samples and laboratory controls. Samples stored as DBS for 2 or more years at ambient temperature showed a significant loss of sensitivity that increased with time; after 10 years only 10% samples with parasite densities > 1,000 parasites/μL were detectable by nested polymerase chain reaction (PCR). Conversely, DBS and extracted DNA stored at -20°C showed no loss of sensitivity with time. Samples with low parasite densities amplified more successfully with saponin/Chelex compared with spin-column-based extraction, though the latter method performed better on samples with higher parasite densities stored for 2 years at ambient temperature. DNA extracted via both methods was stable after 20 freeze-thaw cycles. Our results suggest that DBS should be stored at -20°C or extracted immediately...Continue Reading

References

Oct 2, 2008·Malaria Journal·Patrick H CorranEleanor Riley
Nov 30, 2012·The New England Journal of Medicine·Jane AchanMoses R Kamya

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Citations

Jul 9, 2016·Journal of Pharmaceutical and Biomedical Analysis·Yeliz EnderleJürgen Burhenne
Jul 4, 2018·The American Journal of Tropical Medicine and Hygiene·Mark D Lim
Oct 11, 2017·The American Journal of Tropical Medicine and Hygiene·Shereen KatrakBryan Greenhouse
Feb 10, 2018·Malaria Journal·Christian NsanzabanaIveth J González
May 17, 2019·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Michelle S HsiangBryan Greenhouse
Apr 4, 2017·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Nikhil RanadiveMichelle S Hsiang
Mar 27, 2019·Scientific Reports·Ailie RobinsonColin J Sutherland
Jul 16, 2020·Biology Methods and Protocols·Neta SimonWhitney E Harrington

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