Manger Ronald, Woodle Doug, Berger Andrew, Dickey Robert W, Jester Edward, Yasumoto Takeshi, Lewis Richard, Hawryluk Timothy, Hungerford James
J AOAC Int. 2014 Mar-Apr;97(2):299-306. doi: 10.5740/jaoacint.sgemanger.
Ciguatoxins are potent neurotoxins with a significant public health impact. Cytotoxicity assays have allowed the most sensitive means of detection of ciguatoxin-like activity without reliance on mouse bioassays and have been invaluable in studying outbreaks. An improvement of these cell-based assays is presented here in which rapid flow cytometric detection of ciguatoxins and saxitoxins is demonstrated using fluorescent voltage sensitive dyes. A depolarization response can be detected directly due to ciguatoxin alone; however, an approximate 1000-fold increase in sensitivity is observed in the presence of veratridine. These results demonstrate that flow cytometric assessment of ciguatoxins is possible at levels approaching the trace detection limits of our earlier cytotoxicity assays, however, with a significant reduction in analysis time. Preliminary results are also presented for detection of brevetoxins and for automation and throughput improvements to a previously described method for detecting saxitoxins in shellfish extracts.
西加毒素是具有重大公共卫生影响的强效神经毒素。细胞毒性测定法已成为检测西加毒素样活性的最灵敏方法，无需依赖小鼠生物测定法，并且在研究疫情爆发方面具有重要价值。本文介绍了这些基于细胞的测定法的一种改进方法，其中使用荧光电压敏感染料展示了对西加毒素和石房蛤毒素的快速流式细胞术检测。仅西加毒素就能直接检测到去极化反应；然而，在藜芦碱存在的情况下，灵敏度可提高约1000倍。这些结果表明，流式细胞术评估西加毒素的水平可接近我们早期细胞毒性测定法的痕量检测限，而且分析时间显著缩短。还展示了检测短裸甲藻毒素的初步结果，以及对先前描述的检测贝类提取物中石房蛤毒素方法的自动化和通量改进。
