Venkatesan C, Sarathi M, Balasubramanian G, Saravanan A, Vimal S, Madan N, Majeed S Abdul, Raj N Sundar, Hameed A S Sahul, Babu V Sarath
Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam, Vellore, Tamil Nadu, India.
Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam, Vellore, Tamil Nadu, India Department of Biology, Lakehead University, Thunder Bay, ON, Canada.
Hum Exp Toxicol. 2014 Jul;33(7):772-82. doi: 10.1177/0960327113511474. Epub 2013 Dec 3.
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.
开发了一种夹心酶联免疫吸附测定法（ELISA），用于检测在不同时间间隔（0、2、4、6、8和12小时，直至24小时）向小鼠注射眼镜蛇毒液后，其各种组织（脑、心脏、肺、肝脏、脾脏、血液、肾脏及注射部位组织）中印度眼镜蛇（Naja naja naja）的毒液。全毒液抗血清或单个毒液蛋白抗血清（14、29、65、72和99 kDa）可通过蛋白质印迹法和ELISA识别印度眼镜蛇毒液，并且抗体效价也通过ELISA进行测定。用兔制备的抗眼镜蛇毒抗血清在治疗后的不同时间间隔显著中和了注射毒液小鼠的毒性。该测定法可检测组织匀浆中低至2.5 ng/ml的印度眼镜蛇毒液水平，并且在注射毒液后24小时内均可检测到毒液。在脑、心脏、肺、肝脏、脾脏、肾脏、咬伤部位组织和血液中均检测到了毒液。如在小鼠中观察到的那样，咬伤部位组织中的毒液浓度最高，而脑中的毒液浓度最低。在肝脏、脾脏、肾脏、心脏和肺中发现了适量的毒液。讨论了基于这种ELISA技术开发一种对临床医生有用的简单、快速且具有物种特异性的诊断试剂盒。
