Detection of polymerase chain reaction-amplified malarial DNA in infected blood and individual mosquitoes.
Chelex treatment of Plasmodium falciparum and P. berghei infected tissues, in lieu of organic extraction, was followed directly by polymerase chain reaction amplification of primed circumsporozoite gene sequences. The amplified DNA products were detected in stained gels and hybridization blots of extracts from individual infected mosquitoes and dissected mosquito tissues as well as small volumes of infected blood. Parasite development, within the mosquito midgut and salivary gland, was also monitored as a function of time post infectious blood meal. The temporal presence of amplifiable circumsporozoite gene sequences in the infected mosquito midgut lumen, midgut endothelium, and salivary glands corresponded directly to the visual identification of ookinetes, oocysts, and salivary gland sporozoites, respectively.
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore 21201.
M E
ME
Schriefer
J B
JB
Sacci
R A
RA
Wirtz
A F
AF
Azad
D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D000953:Antigens, Protozoan / Q000235:genetics; D001483:Base Sequence; D016054:DNA, Protozoan / Q000032:analysis* / Q000097:blood; D007475:Ion Exchange Resins; D008288:Malaria / Q000469:parasitology; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010961:Plasmodium / Q000302:isolation & purification*; D010962:Plasmodium berghei / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D010963:Plasmodium falciparum / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011137:Polystyrenes; D011145:Polyvinyls; D015800:Protozoan Proteins; D012680:Sensitivity and Specificity
D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D007475:Ion Exchange Resins; D009838:Oligodeoxyribonucleotides; D011137:Polystyrenes; D011145:Polyvinyls; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan; C006960:chelex