Immunohistochemical demonstration of bleeding in decomposed bodies by using anti-glycophorin A monoclonal antibody.

The usefulness of glycophorin A (GPA) as a marker of bleeding was investigated in decomposed bodies by using anti-human GPA monoclonal antibody immunohistochemically. Ninety-one specimens consisting of 37 skin and 54 muscle specimens were obtained from 21 autopsy cases with various degree of decomposition, which ranged from 12 h up to 2-3 months after death. The presence or absence of the bleeding in the specimens was evaluated macroscopically and was divided as follows: (1) specimens without bleeding (31 specimens), (2) specimens with bleeding (15 specimens), and (3) suspect specimens (45 specimens), in which the bleeding was not clear. By a peroxidase-labeled streptavidin-biotin method, positive reaction products for GPA were observed only within the blood vessels in the specimens without bleeding. On the other hand, in the specimens with bleeding, positive reaction products for GPA were seen not only within the blood vessels but also the extravascular tissues. Therefore, a specimen can be diagnosed as bleeding when GPA is distributed both within blood vessels and tissue outside the vessels. In application of GPA to 45 suspect specimens, 42 specimens (93%) were distinguished from the specimens with bleeding or without bleeding. These results prove that GPA is very useful as a marker of bleeding. The detection of GPA by the immunohistochemical method will help to differentiate between bleeding and hemoglobin (Hb) diffusion from blood vessels in a decomposed body.