The in vivo Pig-a assay: A report of the International Workshop On Genotoxicity Testing (IWGT) Workgroup

Mutat Res Genet Toxicol Environ Mutagen. 2015 May 1:783:23-35. doi: 10.1016/j.mrgentox.2014.09.007. Epub 2014 Sep 26.


The in vivo Pig-a assay uses flow cytometry to measure phenotypic variants for antibody binding to cell surface glycosylphosphatidylinositol (GPI)-anchored proteins. There is good evidence suggesting that the absence of antibody binding is the result of a mutation in the endogenous X-linked Pig-a gene, which forms the rationale for the assay. Although the assay has been performed with several types of hematopoietic cells and in a variety of mammalian species, including humans, currently it is optimized only for measuring CD59-deficient (presumed Pig-a mutant) erythrocytes in the peripheral blood of rats. An expert workgroup formed by the International Workshop on Genotoxicity Testing considered the state of assay development and the potential of the assay for regulatory use. Consensus was reached on what is known about the Pig-a assay and how it should be conducted, and recommendations were made on additional data and refinements that would help to further enhance the assay for use in hazard identification and risk assessment.

Keywords: CD59; Flow cytometry; Glycosylphosphatidylinositol; Mutation; Red blood cells; Reticulocytes.

MeSH terms

  • Anemia, Hemolytic* / metabolism
  • Anemia, Hemolytic* / pathology
  • Animals
  • Antibodies / chemistry
  • Education
  • Erythrocytes* / metabolism
  • Erythrocytes* / pathology
  • Flow Cytometry* / methods
  • Flow Cytometry* / standards
  • Hemoglobinuria* / metabolism
  • Hemoglobinuria* / pathology
  • Humans
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Mutation*
  • Rats


  • Antibodies
  • Membrane Proteins
  • phosphatidylinositol glycan-class A protein

Supplementary concepts

  • CD59 Deficiency