title: Extraction of DNA from malaria-infected erythrocytes using isotachophoresis.
abstract: We demonstrate a technique for purification of nucleic acids from malaria parasites infecting human erythrocytes using isotachophoresis (ITP). We release nucleic acids from malaria-infected erythrocytes by lysing with heat and proteinase K for 10 min and immediately, thereafter, load sample onto a capillary device. We study the effect of temperature on lysis efficiency. We also implement pressure-driven counterflow during ITP extraction to extend focusing time and increase nucleic acid yield. We show that the purified genomic DNA samples are compatible with polymerase chain reaction (PCR) and demonstrate a clinically relevant limit of detection of 0.5 parasites per nanoliter using quantitative PCR.
authors.affiliation: Department of Chemical Engineering, Stanford University, Stanford, California 94305, USA.
authors.forename: Lewis A
authors.initials: LA
authors.lastname: Marshall
authors.forename: Crystal M
authors.initials: CM
authors.lastname: Han
authors.forename: Juan G
authors.initials: JG
authors.lastname: Santiago
mesh_terms: D016054:DNA, Protozoan / Q000302:isolation & purification*; D019286:Endopeptidase K / Q000378:metabolism; D004912:Erythrocytes / Q000469:parasitology*; D006801:Humans; D057625:Isotachophoresis; D008288:Malaria / Q000469:parasitology*; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification; D013696:Temperature
chemical_list: D016054:DNA, Protozoan; D019286:Endopeptidase K