Use of glass beads and CF 11 cellulose for removal of leukocytes from malaria-infected human blood in field settings.
Passage of malaria-infected blood through a two-layered column composed of acid-washed glass beads and CF 11 cellulose removes white cells from parasitized blood. However, because use of glass beads and CF 11 cellulose requires filtration of infected blood separately through these two resins and the addition of ADP, the procedure is time-consuming and may be inappropriate for use in the field, especially when large numbers of blood samples are to be treated. Our modification of this process yields parasitized cells free of contaminating leukocytes, and because of its operational simplicity, large numbers of blood samples can be processed. Our procedure also compares well with those using expensive commercial Sepacell resins in its ability to separate leukocytes from whole blood. As a test of usefulness in molecular biologic investigations, the parasites obtained from the blood of malaria-infected patients using the modified procedure yield genomic DNA whose single copy gene, the circumsporozoite gene, efficiently amplifies by polymerase chain reaction.
Malaria Branch, NCID-CDC, Public Health Service, Atlanta, GA 30333.
I F
IF
Goldman
S H
SH
Qari
J
J
Skinner
S
S
Oliveira
J M
JM
Nascimento
M M
MM
Póvoa
W E
WE
Collins
A A
AA
Lal
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000327:Adsorption; D000818:Animals; D001483:Base Sequence; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002482:Cellulose; D002845:Chromatography / Q000379:methods*; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D005374:Filtration; D017125:Genes, Protozoan; D005898:Glass; D006801:Humans; D007962:Leukocytes / Q000469:parasitology; D016778:Malaria, Falciparum / Q000097:blood* / Q000469:parasitology; D016780:Malaria, Vivax / Q000097:blood* / Q000469:parasitology; D008863:Microspheres; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000235:genetics / Q000302:isolation & purification*; D015539:Platelet Activation; D015800:Protozoan Proteins / Q000235:genetics
D016054:DNA, Protozoan; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan; D000244:Adenosine Diphosphate; D002482:Cellulose