A double antibody sandwich micro-ELISA kit for the rapid diagnosis of snake bite

A micro ELISA assay was established to diagnose systemic poisoning for the rapid administration of specific antivenom. Rabbit anti venom IgG was bound to the solid phase to enable detection of venom from both the Malayan Pit Viper (Agkistrodon rhodostoma) and the Common Cobra (Naja naja). This assay is read visually and takes 35 to 45 minutes to perform. It can detect 15.6 ng/ml of viper venom in 75 minutes and 7.8 ng/ml of cobra venom in 55 minutes. Tests on sera from snake bite patients showed detectable levels of snake venom in the serum even though administration of antivenom was not necessary. Furthermore, results from these clinical cases were obtained in less than 45 minutes. It was found that the most suitable washing media was saline/Tween, the assay could be performed at room temperature and plates stored for 6 months showed no loss of activity.