Accession Number : ADA264323


Title :   Two-Color Flow Cytometric Analysis of Intraerythrocytic Malaria Parasite DNA and Surface Membrane-Associated Antigen in Erythrocytes Infected with Plasmodium falciparum


Corporate Author : WALTER REED ARMY INST OF RESEARCH WASHINGTON DC


Personal Author(s) : Pattanapanyasat, Kovit ; Udomsangpetch, Rachanee ; Webster, H K


Full Text : https://apps.dtic.mil/dtic/tr/fulltext/u2/a264323.pdf


Report Date : Jan 1993


Pagination or Media Count : 7


Abstract : A method for fixation of Plasmodium falciparum infected erythrocytes and solubilization of the erythrocyte membrane with detergent was developed. This method was applied to two-color flow cytometric analysis of both intraerythrocytic (IE) malaria DNA and parasite-derived antigen on the erythrocyte surface membrane. Infected erythrocytes were fixed with 0.025% glutaraldehyde followed by treatment with 1% saponin to gain access to intramembranous components and allow antibody to interact with antigen. DNA of IE parasite was subsequently stained with propidium iodide. Using this procedure cell morphology was well preserved with excellent parasite DNA staining. Using anti-malaria antibodies which recognize ring-infected erythrocyte sur antigen (Pfl55/RESA), we observed that glutaraldehyde-fixed saponin treated infected erythrocytes exhibited a variable immunofluorescence intensity as assessed by both flow cytometry and fluorescence microscopy. Ring-infected cells displayed strong immunofluorescence staining, whereas a weak signal was detected on cells containing schizonts. Simultaneous measurement of parasite DNA and antigen in the infected erythrocyte membrane can facilitate the study of antigen expression in the cell membrane in association with development of IE parasites. Fluorescein, Malaria antigen, Nucleic acid fluorochrome, Propidium iodide, Red blood cell


Descriptors :   *DEOXYRIBONUCLEIC ACIDS , *PARASITES , *MALARIA , *ANTIGENS , *BIOLOGICAL STAINS , *BLOOD CELLS , *ERYTHROCYTES , REPRINTS , MEASUREMENT , DETECTION , FLUORESCENCE , HUMANS , SURFACES , SIGNALS , GAIN , CULTURES(BIOLOGY) , DECAY , PLASMODIUM FALCIPARUM , RINGS , PATIENTS , FLOW , ANTIBODIES , MEMBRANES , DETERGENTS , IODIDES , GLYCOSIDES , PHOSPHATES , NUCLEIC ACIDS , COLORS , QUALITY , MICROSCOPY , PEPTIDES , PROTEINS , MORPHOLOGY , INTENSITY


Subject Categories : Biochemistry
      Medicine and Medical Research
      Microbiology


Distribution Statement : APPROVED FOR PUBLIC RELEASE