Direct PCR amplification and sequence analysis of extrachromosomal Plasmodium DNA from dried blood spots

Acta Trop. 1997 Oct 14;68(1):105-14. doi: 10.1016/s0001-706x(97)00080-6.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA, Protozoan / blood*
  • DNA, Protozoan / chemistry
  • Humans
  • Molecular Sequence Data
  • Plasmodium / genetics*
  • Polymerase Chain Reaction*
  • RNA, Ribosomal / chemistry
  • Sequence Alignment

Substances

  • DNA, Protozoan
  • RNA, Ribosomal