A method for the quantitative assessment of malaria parasite development in organs of the mammalian host.
A non-radioactive PCR method was developed to quantify the development of malaria parasites in the infected host. This was achieved by using Plasmodium genus-specific primers corresponding to the parasite's small subunit ribosomal RNA genes. The quantification of the PCR product was performed by high performance liquid chromatography, and calibration curves were obtained by amplification from defined quantities of purified Plasmodium genomic DNA. Using this method, it was possible to quantify development of P. berghei and P. yoelii blood-stage parasites from blood and brain samples of infected mice, and of hepatic stage parasites, from liver samples of mice infected with different numbers of sporozoites.
Hulier
E
E
U313 INSERM, Paris, France.
Pétour
P
P
Snounou
G
G
Nivez
M P
MP
Miltgen
F
F
Mazier
D
D
Rénia
L
L
D000818:Animals; D001921:Brain / Q000098:blood supply / Q000469:parasitology; D002196:Capillaries / Q000469:parasitology; D016054:DNA, Protozoan / Q000032:analysis* / Q000097:blood; D004912:Erythrocytes / Q000469:parasitology; D005260:Female; D008099:Liver / Q000469:parasitology; D008288:Malaria / Q000469:parasitology*; D051379:Mice; D008815:Mice, Inbred Strains; D010962:Plasmodium berghei / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D010967:Plasmodium yoelii / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D016053:RNA, Protozoan / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D013045:Species Specificity; D013154:Spleen / Q000469:parasitology
D016054:DNA, Protozoan; D016053:RNA, Protozoan; D012335:RNA, Ribosomal