Direct PCR amplification and sequence analysis of extrachromosomal Plasmodium DNA from dried blood spots.
The Plasmodium parasite possesses two extrachromosomal genomes; the mitochondrial genetic element and the extrachromosomal plastid-like DNA. The latter has only been fully described for one culture strain of P. falciparum. In this study, a rapid procedure for amplifying plastid DNA from dried blood spots of blood infected with different malaria species was developed. PCR amplification of a 595 bp fragment within the plastid-like large subunit ribosomal-RNA (LSU-rRNA) gene was achieved using primers derived from the P. falciparum sequence. The PCR product was observed in all Plasmodium species examined. Sequence analysis of amplified products homologous to an LSU-rRNA fragment of the plastid-like extrachromosomal circle revealed extensive conservation between Plasmodium species including P. falciparum, P. vivax, P. malariae and P. berghei.
Tan
T M
TM
Institute of Molecular and Cell Biology, National University of Singapore, Singapore.
Nelson
J S
JS
Ng
H C
HC
Ting
R C
RC
Kara
U A
UA
D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000097:blood* / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D010961:Plasmodium / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal / Q000737:chemistry; D016415:Sequence Alignment
D016054:DNA, Protozoan; D012335:RNA, Ribosomal