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...

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Bibliographic Details
Published in:Acta tropica 1997-10, Vol.68 (1), p.105-114
Main Authors: Tan, T.M.C., Nelson, J.S., Ng, H.C., Ting, R.C.Y., Kara, U.A.K.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Blood spots
DNA, Protozoan - blood
DNA, Protozoan - chemistry
Extrachromosomal plastid
Humans
Malaria
Molecular Sequence Data
PCR amplification
Plasmodium - genetics
Polymerase Chain Reaction
RNA, Ribosomal - chemistry
Sequence Alignment
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Summary: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.
ISSN:0001-706X
1873-6254
DOI:10.1016/S0001-706X(97)00080-6