Detection and neutralization of cobra venom using rabbit antiserum in experimental envenomated mice

• Published in: Human & experimental toxicology Vol. 33; no. 7; pp. 772 - 782
• Main Authors: Venkatesan, C, Sarathi, M, Balasubramanian, G, Saravanan, A, Vimal, S, Madan, N, Majeed, S Abdul, Raj, N Sundar, Hameed, AS Sahul, babu, V Sarath
• Format: Journal Article
• Language: English
• Published: London, England SAGE Publications 01.07.2014; Sage Publications; Sage Publications Ltd
• Subjects: Animal poisons toxicology. Antivenoms; Animals; Antibodies, Neutralizing - pharmacology; Antivenins - pharmacology; Biological and medical sciences; Blotting, Western; Disease Models, Animal; Elapid Venoms - blood; Elapid Venoms - immunology; Elapid Venoms - toxicity; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay - methods; Enzymes; Lethal Dose 50; Male; Medical sciences; Mice; Monoclonal antibodies; Naja; Naja naja naja; Predictive Value of Tests; Rabbits; Rodents; Snake Bites - blood; Snake Bites - diagnosis; Snake Bites - drug therapy; Snake Bites - immunology; Snakes; Time Factors; Tissues; Toxicology; Venom; cobra; cocktail antiserum; antiserum; Naja naja naja; ELISA; Rodentia; Rabbit; Animal origin; Lagomorpha; Enzyme immunoassay; Ophidia; Vertebrata; Mammalia; Antiserum; Mouse; Animal; Venom; Reptilia; Diagnosis; Detection; Naja naja; ELISA assay; Neutralization
• ISSN: 0960-3271; 1477-0903
• DOI: 10.1177/0960327113511474

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect the venom of Indian cobra (Naja naja naja) in various tissues (brain, heart, lungs, liver, spleen, blood, kidneys, and tissue at the site of injection) of mice after cobra venom injected at different time intervals (0, 2, 4, 6, 8, and 12 h intervals up to 24 h). Whole venom antiserum or individual venom protein antiserum (14, 29, 65, 72, and 99 kDa) could recognize N. n. naja venom by Western blotting and ELISA, and antibody titer was also assayed by ELISA. Antiserum raised against cobra venom in rabbit significantly neutralized the toxicity of venom-injected mice at different time intervals after treatment. The assay could detect N. n. naja venom levels up to 2.5 ng/ml of tissue homogenate, and the venom was detected up to 24 h after venom injection. Venom was 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-specific diagnostic kit based on this ELISA technique useful to clinicians is discussed.