Demonstration of calicivirus in human faeces by immunosorbent and immunogold-labelling electron microscopy methods

Abstract

Immunosorbent electron microscopy (ISEM) and immuno-gold staining (IGS) electron microscopy methods have been applied to human faeces, shown by direct electron microscopy (EM) to contain calicivirus. Caliciviruses were successfully trapped on grids coated with positive rabbit or human anti-sera against calicivirus, but not with negative sera. Caliciviruses were specifically labelled with gold particles, when treated with positive rabbit or human antisera against calicivirus followed by protein A-gold and goat anti-rabbit or anti-human IgG gold conjugates in an indirect method in suspension. Goat anti-human IgM gold complexes did not react with the available antisera. Attempts to label caliciviruses trapped on the grids were unsuccessful. No cross-reactions were observed with Norwalk agent-like particles, hepatitis A virus or poliovirus type 3 by any of the methods. The results indicate that ISEM and IGS may be useful techniques for detection and identification of small viruses present in low concentrations in faeces.