RT Journal Article SR Electronic T1 Whole genome sequencing of Plasmodium falciparum from dried blood spots using selective whole genome amplification JF bioRxiv FD Cold Spring Harbor Laboratory SP 067546 DO 10.1101/067546 A1 Samuel O. Oyola A1 Cristina V. Ariani A1 William L. Hamilton A1 Mihir Kekre A1 Lucas N. Amenga-Etego A1 Anita Ghansah A1 Gavin G. Rutledge A1 Seth Redmond A1 Magnus Manske A1 Dushyanth Jyothi A1 Chris G. Jacob A1 Thomas D. Otto A1 Kirk Rockett A1 Chris I. Newbold A1 Matthew Berriman A1 Dominic P. Kwiatkowski YR 2016 UL http://biorxiv.org/content/early/2016/08/11/067546.abstract AB Translating genomic technologies into healthcare applications for the malaria parasite Plasmodium falciparum has been limited by the technical and logistical difficulties of obtaining high quality clinical samples from the field. Sampling by dried blood spot (DBS) finger-pricks can be performed safely and efficiently with minimal resource and storage requirements compared with venous blood (VB). Here, we evaluate the use of selective whole genome amplification (sWGA) to sequence the P. falciparum genome from clinical DBS samples, and compare the results to current methods using leucodepleted VB. Parasite DNA with high (> 95%) human DNA contamination was selectively amplified by Phi29 polymerase using short oligonucleotide probes of 8-12 mers as primers. These primers were selected on the basis of their differential frequency of binding the desired (P. falciparum DNA) and contaminating (human) genomes. Using sWGA method, we sequenced clinical samples from 156 malaria patients, including 120 paired samples for head-to-head comparison of DBS and leucodepleted VB. Greater than 18-fold enrichment of P. falciparum DNA was achieved from DBS extracts. The parasitaemia threshold to achieve >5x coverage for 50% of the genome was 0.03% (40 parasites per 200 white blood cells). Over 99% SNP concordance between VB and DBS samples was achieved after excluding missing calls. The sWGA methods described here provide a reliable and scalable way of generating P. falciparum genome sequence data from DBS samples. Our data indicate that it will be possible to get good quality sequence data on most if not all drug resistance loci from the majority of symptomatic malaria patients. This technique overcomes a major limiting factor in P. falciparum genome sequencing from field samples, and paves the way for large-scale epidemiological applications.