AJTMH Transactions of the Royal Society of Tropical Medicine and Hygiene
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Am. J. Trop. Med. Hyg., 72(3), 2005, pp. 249-251
Copyright © 2005 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT: RAPID DNA EXTRACTION FROM ARCHIVE BLOOD SPOTS ON FILTER PAPER FOR GENOTYPING OF PLASMODIUM FALCIPARUM

SÁNDOR BERECZKY, ANDREAS MÅRTENSSON, J. PEDRO GIL, AND ANNA FÄRNERT
Infectious Diseases Unit, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden; Emergency Medicine Unit, Department of Medicine, Kullbergska Hospital, Katrineholm, Sweden

 

ABSTRACT

The practical advantages of sampling and storing blood on filter paper for analyses of human and pathogen genes highlight the need for reliable, sensitive, and cost-effective DNA extraction methods. We describe a new Tris-EDTA (TE) buffer-based method for extraction of DNA from blood dried on filter paper. The method was evaluated against the commonly used methanol and Chelex® methods, regarding polymerase chain reaction detection of Plasmodium falciparum parasites from samples stored for 1–2 years. The sensitivity of detection was dependent on the parasite density and type of filter paper. For 3MM® Whatman filter paper, the sensitivity was 100%, 73%, and 93% for the TE, methanol, and Chelex® methods, respectively. For the longer stored 903® Schleicher & Schuell filter paper, the sensitivity was 93%, 73%, and 0%, respectively. This rapid, simple, and inexpensive extraction method generated superior results from archived specimens compared with the two standard methods and may represent a useful tool in molecular epidemiologic studies.

Received June 2, 2004. Accepted for publication October 18, 2004.

Acknowledgements: We thank Hanna-Stina Hanson for constructive discussions, Professor Marita Troye-Blomberg and Dr. Johan Strömberg-Nörklit for providing filter paper samples, and Professor Anders Björkman for advice and comments.

Financial support: This work was part of a project supported by the Swedish International Development Cooperation Agency (SAREC).

Authors’ addresses: Sándor Bereczky, J. Pedro Gil, and Anna Färnert, Infectious Diseases Unit, Karolinska Institutet, Karolinska University Hospital, S-171 76 Stockholm, Sweden, Telephone: 46-8-517-75-281, Fax: 46-8-517-76-740, E-mail: sandor.bereczky{at}medks.ki.se. Andreas Mårtensson, Infectious Diseases Unit, Karolinska Institutet, Karolinska University Hospital, S-171 76 Stockholm, Sweden and Emergency Medicine Unit, Department of Medicine, Kullbergska Hospital, Katrineholm, Sweden.




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