A method for the purification of Leishmania promastigotes from infected phlebotomine sandflies.
We describe a method for the purification of Leishmania promastigotes, isolated from infected sandflies (Lutzomyia longipalpis) using a discontinuous density centrifugation gradient (Percoll/Homem). The sandflies, infected seven days previously with Leishmania donovani chagasi or Leishmania mexicana mexicana from culture, were homogenized and centrifuged on a Percoll discontinuous gradient. Five interface bands were formed, and most of the promastigotes settled out at the interface between the (30% and 40%) Percoll concentrations. An extraction of 3.5 x 10(4) promastigotes from 90 female flies was achieved using this technique.
Stephenson
G
G
Department of Medical Entomology, Liverpool School of Tropical Medicine, U.K.
Stokes
R W
RW
Ward
R D
RD
D000818:Animals; D002499:Centrifugation, Density Gradient; D007891:Leishmania / Q000302:isolation & purification*; D007893:Leishmania donovani / Q000302:isolation & purification; D007894:Leishmania mexicana / Q000302:isolation & purification; D007896:Leishmaniasis / Q000469:parasitology; D010691:Phlebotomus / Q000469:parasitology*; D014020:Tissue Extracts / Q000032:analysis
D014020:Tissue Extracts