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Zeitschrift für Parasitenkunde

, Volume 66, Issue 3, pp 273–280 | Cite as

The use of percoll gradients, elutriator rotor elution, and mithramycin staining for the isolation and identification of intraerythrocytic stages ofPlasmodium berghei

  • Lorenz Rüssmann
  • Albrecht Jung
  • Hans-G. Heidrich
Original Investigations

Abstract

Intraerythrocytic parasites ofPlasmodium vinckei andPlasmodium berghei were separated according to their developmental stages using discontinuous Percoll gradients. Contaminating nucleated blood cells such as leukocytes were removed by elutriation centrifugation. The stages were unequivocally identified in smears using a newly developed DNA-specific staining procedure with mithrmycin and fluorescence microscopy. This stain can also be used to detect parasites in human blood of very low parasitemias. The combination of methods described has many possible applications in immunologic and biochemical parasite research.

Keywords

Malaria Infected Erythrocyte Percoll Gradient Gradient Fraction Plasmodium Berghei 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Eling W (1977) Ficoll fractionation for the separation of parasitized erythrocytes from malaria infected blood. Bull WHO 57: 105–114Google Scholar
  2. Eugui EM, Allison AC (1979) Separation of erythrocytes infected with murine malaria parasites in metrizamide gradients. Parasitology 79: 267–275CrossRefPubMedGoogle Scholar
  3. Heidrich HG (1970) Isolation of pure inner and outer mitochondrial membranes in preparative scale. In: Advances in automated chemistry: proceedings of the international Technicon congress 1969, MEDIAD-Technicon, Tarrytown-New York pp 347–350Google Scholar
  4. Heidrich HG, Rieckmann KH (1980) Free-flow electrophoresis separation of intracellular parasites (Plasmodium falciparum) from culture. Eur J Cell Biol 22: 516Google Scholar
  5. Heidrich HG, Rüssmann L, Bayer B, Jung A (1979) Free-flow electrophoresis for the separation of malaria infected and non-infected mouse erythrocytes and for isolation of free parasites (Plasmodium vinckei). Z Parasitenkd 58: 151–159CrossRefPubMedGoogle Scholar
  6. Howard RJ, Battye FL (1979)Plasmodium berghei — infected red cells sorted according to DNA stain. Parasitology 78: 263–270CrossRefPubMedGoogle Scholar
  7. Hyman BC, MacInnis AJ (1979) Rapid detection of malaria and other blood stream parasites by fluorescence microscopy with 4 6 diamidino-2-phenylindole (DAP). J Parasitol 65: 421–425CrossRefPubMedGoogle Scholar
  8. Kilejian A (1980) Stage-specific proteins and glycoproteins ofPlasmodium falciparum: identification of antigens unique to schizonts and merozoites. Proc Natl Acad Sci USA 77: 3695–3699CrossRefPubMedPubMedCentralGoogle Scholar
  9. Lunde MM, Powers KS (1976) The preparation of malaria hemagglutination antigen. Ann Trop Med Parasitol 70: 283–291CrossRefPubMedGoogle Scholar
  10. McAllister RO, Gordon DM (1976) Schizont-infected cell enrichment in rodent malaria. J Parasitol 62: 664–669CrossRefGoogle Scholar
  11. McEven CR, Juhos ET, Stallard RW, Schnell JV, Siddiqui WA, Geimann QM (1971) Centrifugal elutriation for the removal of leukocytes from malaria-infected monkey blood. J Parasitol 57: 887–890CrossRefGoogle Scholar
  12. Miller LH, Chien S (1971) Density distribution of red cells infected byPlasmodium knowlesi andPlasmodium coatneyi. Exp Parasitol 29: 451–456CrossRefPubMedGoogle Scholar
  13. Mrema JE, Campbell GH, Miranda R, Jamarillo AL, Rieckmann KH (1979) Concentration and separation of erythrocytes infected withPlasmodium falciparum by gradient centrifugation. Bull WHO 57: 133–138PubMedPubMedCentralGoogle Scholar
  14. Pasvol G, Wilson RJ, Smalley ME, Brown J (1978) Separation of viable schizont-infected red cells ofPlasmodium falciparum from human blood. Ann Trop Med Parasitol 72: 87–88CrossRefPubMedGoogle Scholar
  15. Pertoft H, Laurent TC (1979) Isopycnic separation of cells and cell organelles by centrifugation in modified colloidal silica gradients. In: Catsimpolas N (ed) Methods of cell separation. Plenum, New York, pp 25–56Google Scholar
  16. Reese RT, Langreth SG, Trager W (1979) Isolation of stages of the human parasitePlasmodium falciparum from culture and from blood. Bull WHO 57: 53–61PubMedPubMedCentralGoogle Scholar
  17. Rowley PT, Siddiqui WA, Geiman OM (1967) Separation of malarial parasites according to age by density gradient centrifugation. J Lab Clin Med 70: 933–937PubMedGoogle Scholar
  18. Sanderson RJ, Bird KE (1977) Cell separation by counterflow centrifugation. Methods Cell Biol 15: 1–14CrossRefPubMedGoogle Scholar
  19. Siddiqui WA, Kan SC, Kramer K, Richmond-Crum SM (1979) In vitro production and partial purification ofPlasmodium falciparum antigen. Bull WHO 57: 75–82PubMedPubMedCentralGoogle Scholar
  20. Sodeman TM (1970) The use of fluorochromes for the detection of malaria parasites. Am J Trop Med Hyp 19: 40–45Google Scholar
  21. Tosta CE, Sedegah M, Henderson DC, Wedderburn N (1980)Plasmodium yoelii andPlasmodium berghei: isolation of infected erythrocytes from blood by colloidal silica gradient centrifugation. Exp Parasitol 50: 7–15CrossRefPubMedGoogle Scholar
  22. Ward DC, Reich E, Goldberg IH (1965) Base specificity in the interaction of polynucleotides with antibiotic drugs. Science 149: 1259–1263CrossRefPubMedGoogle Scholar
  23. Williamson J, Cover B (1966) Separation of blood-cell-free trypanosomes and malaria parasites on a sucrose gradient. Trans R Soc Trop Med Hyg 60: 425–427CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Lorenz Rüssmann
    • 1
  • Albrecht Jung
    • 2
  • Hans-G. Heidrich
    • 1
  1. 1.Max-Planck-Institut für BiochemieMartinsried bei MünchenGermany
  2. 2.Physiologisch-chemisches InstitutUniversity of TübingenTübingenGermany

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