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

• 412 Accesses
• 9 Citations
• 3 Altmetric
• Explore all metrics

Abstract

Intraerythrocytic parasites of_Plasmodium vinckei_ and_Plasmodium 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.

Access this article

Log in via an institution

Subscribe and save

• Starting from 10 chapters or articles per month
• Access and download chapters and articles from more than 300k books and 2,500 journals
• Cancel anytime View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

• Eling W (1977) Ficoll fractionation for the separation of parasitized erythrocytes from malaria infected blood. Bull WHO 57: 105–114
  Google Scholar
• Eugui EM, Allison AC (1979) Separation of erythrocytes infected with murine malaria parasites in metrizamide gradients. Parasitology 79: 267–275
  Article CAS Google Scholar
• 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 347–350
• Heidrich HG, Rieckmann KH (1980) Free-flow electrophoresis separation of intracellular parasites (Plasmodium falciparum) from culture. Eur J Cell Biol 22: 516
  Google Scholar
• 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: 151–159
  Article CAS Google Scholar
• Howard RJ, Battye FL (1979)Plasmodium berghei — infected red cells sorted according to DNA stain. Parasitology 78: 263–270
  Article CAS Google Scholar
• 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: 421–425
  Article CAS Google Scholar
• Kilejian A (1980) Stage-specific proteins and glycoproteins of_Plasmodium falciparum_: identification of antigens unique to schizonts and merozoites. Proc Natl Acad Sci USA 77: 3695–3699
  Article CAS Google Scholar
• Lunde MM, Powers KS (1976) The preparation of malaria hemagglutination antigen. Ann Trop Med Parasitol 70: 283–291
  Article CAS Google Scholar
• McAllister RO, Gordon DM (1976) Schizont-infected cell enrichment in rodent malaria. J Parasitol 62: 664–669
  Article Google Scholar
• 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: 887–890
  Article Google Scholar
• Miller LH, Chien S (1971) Density distribution of red cells infected by_Plasmodium knowlesi_ and_Plasmodium coatneyi_. Exp Parasitol 29: 451–456
  Article CAS Google Scholar
• Mrema JE, Campbell GH, Miranda R, Jamarillo AL, Rieckmann KH (1979) Concentration and separation of erythrocytes infected with_Plasmodium falciparum_ by gradient centrifugation. Bull WHO 57: 133–138
  CAS PubMed Google Scholar
• Pasvol G, Wilson RJ, Smalley ME, Brown J (1978) Separation of viable schizont-infected red cells of_Plasmodium falciparum_ from human blood. Ann Trop Med Parasitol 72: 87–88
  Article CAS Google Scholar
• 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 25–56
  Google Scholar
• Reese RT, Langreth SG, Trager W (1979) Isolation of stages of the human parasite_Plasmodium falciparum_ from culture and from blood. Bull WHO 57: 53–61
  PubMed Google Scholar
• Rowley PT, Siddiqui WA, Geiman OM (1967) Separation of malarial parasites according to age by density gradient centrifugation. J Lab Clin Med 70: 933–937
  CAS PubMed Google Scholar
• Sanderson RJ, Bird KE (1977) Cell separation by counterflow centrifugation. Methods Cell Biol 15: 1–14
  Article CAS Google Scholar
• Siddiqui WA, Kan SC, Kramer K, Richmond-Crum SM (1979) In vitro production and partial purification of_Plasmodium falciparum_ antigen. Bull WHO 57: 75–82
  PubMed Google Scholar
• Sodeman TM (1970) The use of fluorochromes for the detection of malaria parasites. Am J Trop Med Hyp 19: 40–45
  Article CAS Google Scholar
• Tosta CE, Sedegah M, Henderson DC, Wedderburn N (1980)Plasmodium yoelii and_Plasmodium berghei_: isolation of infected erythrocytes from blood by colloidal silica gradient centrifugation. Exp Parasitol 50: 7–15
  Article CAS Google Scholar
• Ward DC, Reich E, Goldberg IH (1965) Base specificity in the interaction of polynucleotides with antibiotic drugs. Science 149: 1259–1263
  Article CAS Google Scholar
• 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: 425–427
  Article Google Scholar

Download references

Author information

Authors and Affiliations

1. Max-Planck-Institut für Biochemie, D-8033, Martinsried bei München, Germany
   Lorenz Rüssmann & Hans-G. Heidrich
2. Physiologisch-chemisches Institut, University of Tübingen, D-7400, Tübingen, Germany
   Albrecht Jung

Authors

1. Lorenz Rüssmann
2. Albrecht Jung
3. Hans-G. Heidrich

Rights and permissions

About this article

Cite this article

Rüssmann, L., Jung, A. & Heidrich, HG. The use of percoll gradients, elutriator rotor elution, and mithramycin staining for the isolation and identification of intraerythrocytic stages of_Plasmodium berghei_.Z. Parasitenkd. 66, 273–280 (1982). https://doi.org/10.1007/BF00925344

Download citation

• Received: 17 February 1981
• Published: 01 October 1982
• Issue date: October 1982
• DOI: https://doi.org/10.1007/BF00925344

Keywords