Audebert F, Grosselet O, Sabouraud A, Bon C
Unité des Venins, Institut Pasteur, Paris, France.
J Anal Toxicol. 1993 Jul-Aug;17(4):236-40. doi: 10.1093/jat/17.4.236.
We describe an enzyme-linked immunosorbent assay (ELISA) to quantitate venom antigens in human serum and urine, and thus to help evaluate the severity of envenomation due to viper bites. This assay, which is performed with commercially available polyclonal Fab'2s in a double-sandwich method, is rapid, simple, and specific for antigens of European vipers (Vipera aspis, Vipera berus, and Vipera ammodytes). No cross-reactivity was observed with other snake venoms or human serum proteins. It showed a good linear response over a wide range of concentrations of venom antigens (from 1 to 100 ng/mL). It was very sensitive, with detection limits of 7 and 2 ng/mL for Vipera aspis venom in serum and urine, respectively. This ELISA is also easily reproducible; the coefficients of variation determined at different concentrations of venom (50, 25, and 5 ng/mL) did not exceed 10% in serum and 14% in urine samples collected from different donors. This test was applied to determine the concentrations of venom in the serum of patients bitten by a viper in France and to follow its elimination as a function of time. The method is adaptable to other venoms by using other specific immunoglobulins.
我们描述了一种酶联免疫吸附测定法（ELISA），用于定量人血清和尿液中的毒液抗原，从而有助于评估蝰蛇咬伤所致中毒的严重程度。该测定法采用市售多克隆Fab'2以双夹心方法进行，快速、简单，且对欧洲蝰蛇（角蝰、极北蝰和矛头蝮）的抗原具有特异性。未观察到与其他蛇毒或人血清蛋白的交叉反应。在毒液抗原的广泛浓度范围（1至100 ng/mL）内，它显示出良好的线性响应。它非常灵敏，血清和尿液中角蝰毒液的检测限分别为7和2 ng/mL。这种ELISA也易于重复；在不同浓度毒液（50、25和5 ng/mL）下测定的变异系数，在来自不同供体的血清中不超过10%，在尿液样本中不超过14%。该试验用于测定法国被蝰蛇咬伤患者血清中的毒液浓度，并追踪其随时间的消除情况。通过使用其他特异性免疫球蛋白，该方法可适用于其他毒液。
