Short report: Rapid DNA extraction from archive blood spots on filter paper for genotyping of Plasmodium falciparum.
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.
Bereczky
Sándor
S
Infectious Diseases Unit, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden. sandor.bereczky@medks.ki.se
Mårtensson
Andreas
A
Gil
J Pedro
JP
Färnert
Anna
A
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics; D016054:DNA, Protozoan / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D005838:Genotype; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D010963:Plasmodium falciparum / Q000235:genetics* / Q000302:isolation & purification; D015800:Protozoan Proteins / Q000235:genetics; D012680:Sensitivity and Specificity
D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D015800:Protozoan Proteins; C065974:merozoite surface protein 2, Plasmodium