DETECTION AND NEUTRALIZATION OF VENOM OF INDIAN COBRA BY RABBIT ANTISERUM IN EXPERIMENTAL ENVENOMATION USING MICE

Author(s): C Venkatesan*, M Sarathi Mani, G Balasubramanaiyan, Jhon Thomas, A Saravanan, V Sarath babu, AS Sahu Hameed, Urban JAD Souza

Abstract

A sandwich enzyme linked immunosorbent assay (ELISA) was developed to detect Indian cobra (Naja naja naja) venom in various organs (brain, heart, lungs, liver, spleen, blood, site of injection and kidneys) as well as tissue at the site of injection of mice, at various time intervals (0, 2, 4, 6, 8 and 12 h intervals up to 24 h) after venom injection. Antiserum significantly neutralized venom levels in serum and tissue samples. Whole venom antiserum or individual venom protein anti serum (14 kDa, 29 kDa, 65 kDa, 72 kDa and 99 kDa) of venom could recognize Naja naja venom by Western blotting and ELISA, and Naja naja naja venom presented antibody titer when assayed by ELISA. The assay could detect Naja naja naja venom levels up to 2.5 ng/ml of tissue homogenate and the venom was detected up to 24 h after venom injection. A highly sensitive and species-specifc microtitre ELISA was also developed to detect venoms of four medically important Indian snakes (Naja naja naja) in autopsy specimens of mice. Venoms were detected in brain, heart, lungs, liver, spleen, kidneys, tissue at the bite area and blood. As observed in mice, tissue at the site of bite area showed the highest concentration of venom and the brain showed the least. Moderate amounts of venoms were found in liver, spleen, kidneys, heart and lungs. Development of a simple, rapid and species-specifc diagnostic kit based on this ELISA technique useful to clinicians is discussed.

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