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Related Experiment Videos

A colorimetric assay for trypanosome viability and metabolic function

J A Ellis1, W R Fish, M Sileghem

  • 1International Laboratory for Research on Animal Diseases, Nairobi, Kenya.

Veterinary Parasitology
|October 1, 1993
PubMed
Summary

We adapted the MTT assay for African trypanosome viability. This sensitive method quantifies parasites and can screen drugs and host factors affecting their growth.

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Area of Science:

  • Parasitology
  • Biochemistry
  • Cell Biology

Background:

  • African trypanosomes cause significant human and animal diseases.
  • Assessing parasite viability is crucial for drug discovery and understanding host-parasite interactions.
  • Traditional methods for quantifying trypanosomes are often labor-intensive and lack sensitivity.

Purpose of the Study:

  • To adapt the tetrazolium salt (MTT) colorimetric assay for assessing African trypanosome viability and metabolic function.
  • To evaluate the effect of lymphoid cytokines on trypanosome growth.
  • To establish a sensitive and rapid method for screening growth-modulating factors.

Main Methods:

  • African trypanosomes (T. congolense, T. brucei rhodesianse) were cultured axenically.
  • Serial dilutions of trypanosomes were analyzed using the MTT assay to determine sensitivity.

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  • Trypanosomes were incubated with various lymphoid cytokines (IL-1, IL-3, IL-6, IFN-γ, TNF-α) at different concentrations and time points.

    • Main Results:

    • The MTT assay detected as few as 10(4) trypanosomes.
    • Lymphoid cytokines (human and bovine) showed no significant growth-enhancing or inhibitory effects on trypanosomes.
    • The assay demonstrated high sensitivity, speed, and repeatability compared to traditional counting methods.

    Conclusions:

    • The MTT colorimetric assay is a sensitive and efficient tool for quantifying African trypanosome viability.
    • This adapted assay is suitable for screening potential anti-trypanosomal drugs and host-derived factors.
    • The study provides a valuable method for research on trypanosome biology and disease control.