Use of egg yolk antibody (IgY) as an immunoanalytical tool in the detection of Indian cobra (Naja naja naja) venom in biological samples of forensic origin
Use of egg yolk antibody (IgY) as an immunoanalytical tool in the detection of Indian cobra (Naja naja naja) venom in biological samples of forensic origin
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DOI：
10.1016/j.toxicon.2006.04.011
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发表时间：
2006-08-01
期刊：
影响因子：
2.8
通讯作者：
Sarin, R. K.
中科院分区：
文献类型：
--
作者：
Brunda, G.;Sashidhar, R. B.;Sarin, R. K.
建立了一种基于免疫球蛋白Y（IgY）的间接双抗体夹心酶联免疫吸附试验（ELISA），用于检测法医来源生物样本中的印度眼镜蛇（Naja naja naja）毒液。从鸡蛋黄和兔血清中制备并纯化了多克隆抗体。眼镜蛇毒被夹在固定化亲和纯化的IgY和兔IgG之间。眼镜蛇毒的检测浓度范围为0.1 ~ 300 ng。校准曲线基于线性回归分析（y = 0.2581x + 0.4375，r（2）= 0.9886）。发现测定的检测限为0.1 ng。观察到批间（n = 6）和批内（n = 6）不同浓度工作范围的变异系数（CV）小于10%。当将不同浓度（0.002、0.1、0.2、1和2 μ g）的眼镜蛇毒掺入混合的正常人血清（ml（-1））中时，发现毒液的回收率在80- 99%的范围内。在0.1-1000 ng浓度范围内，与金环蛇毒和蝰蛇毒无交叉反应。该方法最初通过分析注射眼镜蛇毒液（1.2 mg kg（-1）体重）的实验大鼠的标本（尸检）进行验证。此外，还分析了法医来源的蛇咬伤受害者的人体标本（尸检和活检）。所开发的方法可能在法医实验室中找到诊断应用。(c)2006爱思唯尔有限公司保留所有权利。
An immunoglobulin Y (IgY) based indirect double antibody sandwich enzyme linked immunosorbent assay (ELISA) was developed for the detection of Indian cobra (Naja naja naja) venom in the biological samples of forensic origin. Polyclonal antibodies were raised and purified from chick egg yolk and rabbit serum. The cobra venom was sandwiched between immobilized affinity purified IgY and the rabbit IgG. The detection concentration of cobra venom was in the range of 0.1 to 300 ng. The calibration plot was based on linear regression analysis (y = 0.2581x + 0.4375, r(2) = 0.9886). The limit of detection of the assay was found to be 0.1 ng. The coefficient of variation (CV) of different concentrations of working range in inter (n = 6) and intra-assay (n = 6) was observed to be less than 10%. The recovery of venom was found to be in the range of 80-99%, when different concentrations (0.002, 0.1, 0.2, 1, and 2 mu g) of cobra venom were spiked to pooled normal human serum (ml(-1)). No cross reactivity was observed with krait and viper venom in the immunoassay system in the concentration range of 0.1-1000 ng. The method was initially, validated by analyzing specimens (autopsy) of experimental rats injected with cobra venom (1.2 mg kg(-1) body mass). Further, human specimens (autopsy and biopsy) of snake bite victims of forensic origin were also analyzed. The methodology developed may find diagnostic application in forensic laboratories. (c) 2006 Elsevier Ltd. All rights reserved.