Manger R L, Leja L S, Lee S Y, Hungerford J M, Hokama Y, Dickey R W, Granade H R, Lewis R, Yasumoto T, Wekell M M
U.S. Food and Drug Administration, Seafood Products Research Center, Bothell, WA 98041-3012, USA.
J AOAC Int. 1995 Mar-Apr;78(2):521-7.
Neuroblastoma cells in culture were used to detect sodium channel-specific marine toxins based on an end-point determination of mitochondrial dehydrogenase activity. The assay responds in a dose-dependent manner to ciguatoxins, brevetoxins, and saxitoxins, and delineates the toxic activity as either sodium channel enhancing or sodium channel blocking. The assay responds rapidly to sodium channel activating toxins, allowing dose dependent detection in 4 to 6 h. Brevetoxins can be detected at 250 pg, and purified ciguatoxins are detected in the low picogram and subpicogram levels. The results obtained from cell bioassay of ciguatoxic finfish extracts correlates with those obtained from mouse bioassays. Sodium channel blocking toxins can also be detected with an approximate sensitivity of 20 pg in 24 to 48 h. This cell-based technique is simple, sensitive, demonstrates potential as an alternative to animal testing for sodium channel activating and blocking toxins, and can be automated.
利用培养的神经母细胞瘤细胞，基于线粒体脱氢酶活性的终点测定来检测钠通道特异性海洋毒素。该检测方法对雪卡毒素、短裸甲藻毒素和石房蛤毒素呈剂量依赖性反应，并将毒性活性描述为钠通道增强或钠通道阻断。该检测方法对钠通道激活毒素反应迅速，可在4至6小时内进行剂量依赖性检测。短裸甲藻毒素的检测下限为250皮克，纯化的雪卡毒素可在低皮克和亚皮克水平被检测到。从雪卡毒素中毒鱼类提取物的细胞生物测定中获得的结果与从小鼠生物测定中获得的结果相关。钠通道阻断毒素也可在24至48小时内以约20皮克的灵敏度被检测到。这种基于细胞的技术简单、灵敏，显示出作为钠通道激活和阻断毒素动物试验替代方法的潜力，并且可以实现自动化。
