Division of Toxicology, China Medical University Hospital , Taichung , Taiwan.
Clin Toxicol (Phila). 2014 Mar;52(3):187-91. doi: 10.3109/15563650.2014.887725.
The clinical diagnosis of snakebites is critical and necessary in many parts of the world, especially in Southeastern Asia, where venomous snakebites are a burden on public health. It is difficult to define or recognize the species of venomous snake because of the overlapping clinical manifestations of envenomations. A quick and reliable method for identifying the snake species is necessary. We designed and tested a strip of lateral flow system for the diagnosis of cobra snake bites in Taiwan.
We developed a kit based on an immunochromatographic method for rapid detection of cobra (Naja atra) venom in human serum. The test and control lines composed of 1 mg/ml polyclonal duck antivenom and 0.5 mg/ml goat anti-rabbit immunoglobulin antibody solutions, respectively, were coated on nitrocellulose strips. Colloidal gold was conjugated with rabbit polyclonal anti-cobra venom antibodies. From July 2007 to December 2012, we used the kit to test serum from snakebite patients and to examine the agreement between our rapid test and the currently used sandwich enzyme-linked immunosorbent assay (ELISA).
Our kit was able to detect cobra venom in serum samples in 20 minutes with a detection limit of 5 ng/ml. An absence of cross-reactivity with other non-cobra venoms from Taiwan was noted in vitro. A total of 88 snakebite patients (34 cobra and 54 other non-cobra) were tested. The sensitivity of the strips based on the ELISA results was 83.3% and the specificity was 100%. There was a strong agreement between the results of the ELISA and immunochromatographic strips (κ = 0.868).
This data indicates that an immunochromatographic strip might be suitable for cobra venom detection and could be used as a quick diagnostic tool in cases of N. atra snakebite.
在世界许多地区,包括东南亚,毒蛇咬伤的临床诊断至关重要,因为毒蛇咬伤对公共健康造成了负担。由于毒液中毒的临床表现重叠,确定毒蛇的种类具有一定难度。因此,需要一种快速而可靠的方法来识别蛇的种类。我们设计并测试了一种用于诊断台湾眼镜蛇咬伤的侧向流动系统试纸条。
我们基于免疫层析法开发了一种用于快速检测人血清中眼镜蛇(Naja atra)毒液的试剂盒。检测线和控制线分别包被了 1 mg/ml 的多克隆鸭抗蛇毒血清和 0.5 mg/ml 的山羊抗兔免疫球蛋白抗体溶液。胶体金标记了兔多克隆抗眼镜蛇毒抗体。从 2007 年 7 月至 2012 年 12 月,我们使用该试剂盒检测了蛇咬伤患者的血清,并比较了快速检测法与当前使用的夹心酶联免疫吸附试验(ELISA)之间的一致性。
我们的试剂盒在 20 分钟内能够检测到血清中的眼镜蛇毒液,检测限为 5 ng/ml。体外实验表明,该试剂盒与台湾的其他非眼镜蛇蛇毒无交叉反应。总共检测了 88 名蛇咬伤患者(34 名眼镜蛇咬伤患者和 54 名其他非眼镜蛇咬伤患者)。根据 ELISA 结果,该试纸条的灵敏度为 83.3%,特异性为 100%。ELISA 和免疫层析试纸条的结果具有很强的一致性(κ=0.868)。
这些数据表明,免疫层析试纸条可能适合于检测眼镜蛇毒液,可作为 N. atra 蛇咬伤的快速诊断工具。