The dot blot direct antiglobulin test

Am J Clin Pathol. 1987 Dec;88(6):733-8. doi: 10.1093/ajcp/88.6.733.

Abstract

The direct antiglobulin test (DAT) is the most widely used serologic method to determine whether antibody or complement has bound to red blood cells in vivo. A solid phase DAT, based on the dot immunobinding technic, has been developed (DOT DAT). The solid phase was prepared by attaching anti-human IgG to nitrocellulose membranes. Patients' red blood cells were washed in saline and layered on top of the membranes. After 5 minutes the membranes were washed and the results were read. A positive reaction had a red dot of adherent cells on the membrane, whereas a negative membrane remained white. Good correlation was observed between the DOT DAT and the hemagglutination DAT after testing of 131 patient samples. The primary advantages of the DOT DAT were its stability, simplicity, and objective end point.

MeSH terms

  • Antibodies / analysis
  • Complement System Proteins / analysis
  • Coombs Test*
  • Erythrocyte Membrane / immunology
  • Hemagglutination Tests
  • Humans
  • Immunoassay

Substances

  • Antibodies
  • Complement System Proteins