电压敏感荧光染料在鱼贝类毒素检测中的应用初探

目的建立快速、简便、能反映样品整体毒性的鱼贝类毒素筛查方法。方法以神经母细胞瘤细胞为实验材料,采用电压敏感荧光染料bis-oxonol,根据膝沟藻毒素2,3(GTX2,3)、短裸甲藻毒素(BTX)和河豚鱼毒(TTX)的毒理特点,观察不同浓度毒素作用下细胞荧光的变化。结果 GTX2,3的浓度在2～200ng/ml范围内,TTX的浓度在20～600ng/ml范围内,BTX的浓度在15～400ng/ml范围内,培养体系荧光强度与毒素的浓度之间存在很好的线性关系。结论利用电压敏感荧光染料法可实现对GTX2,3、TTX和BTX的快速检测。

Application of a fluorescent dye method based on changes in membrane potential in detecting shellfish toxins

Objective To develop a rapid and sensitive assay for detecting the common sea food toxins including paralytic shellfish poisoning toxin(PST),tetrodotoxin(TTX) and neurotoxic shellfish poisoning toxin(NST) based on their toxicological character.Methods Neuroblastoma cells were incubated with the uorescent dye bis-oxonol,whose distribution across the membrane was potential-dependent.Changes in membrane potential of the cells induced by gonyautoxins(GTX2,3),brevetoxin(BTX) and TTX were observed respectively,using bis-oxonol.Results Within 2-200nmol/L of GTX2,3 or 20-600nmol/L of TTX,veratridine-induced depolarization was shown to be inhibited by GTX2,3 or TTX in dose-dependent manner.Within 15-400ng/ml,there was a dose-dependent relationship between the NSP-induced depolarization and toxin concentration.Conclusion It was likely to find a rapid,specific,and reliable method with bis-oxonol for detecting GTX2,3,TTX and BTX in sea food.