Interfibio, Nanobiotechnology and Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Paisos Catalans 26, 43007 Tarragona, Spain.
IRTA, Ctra. Poble Nou km 5.5, 43540 Sant Carles de la Ràpita, Spain.
Anal Chem. 2021 Nov 9;93(44):14810-14819. doi: 10.1021/acs.analchem.1c03671. Epub 2021 Oct 26.
The marine toxin tetrodotoxin (TTX) poses a great risk to public health safety due to its severe paralytic effects after ingestion. Seafood poisoning caused by the consumption of contaminated marine species like pufferfish due to its expansion to nonendemic areas has increased the need for fast and reliable detection of the toxin to effectively implement prevention strategies. Liquid chromatography-mass spectrometry is considered the most accurate method, although competitive immunoassays have also been reported. In this work, we sought to develop an aptamer-based assay for the rapid, sensitive, and cost-effective detection of TTX in pufferfish. Using capture-SELEX combined with next-generation sequencing, aptamers were identified, and their binding properties were evaluated. Finally, a highly sensitive and user-friendly hybrid antibody-aptamer sandwich assay was developed with superior performance compared to several assays reported in the literature and commercial immunoassay kits. The assay was successfully applied to the quantification of TTX in pufferfish extracts, and the results obtained correlated very well with a competitive magnetic bead-based immunoassay performed in parallel for comparison. This is one of the very few works reported in the literature of such hybrid assays for small-molecule analytes whose compatibility with field samples is also demonstrated.
海洋毒素河豚毒素 (TTX) 具有严重的麻痹作用，摄入后会对公众健康安全构成极大威胁。由于其分布范围扩大到非流行地区，食用受污染的海洋物种（如河豚鱼）引起的海鲜中毒事件有所增加，这就需要快速、可靠地检测毒素，以便有效地实施预防策略。液相色谱-质谱联用被认为是最准确的方法，尽管也有报道称竞争性免疫分析是一种方法。在这项工作中，我们试图开发一种基于适配体的方法，用于快速、灵敏、经济高效地检测河豚鱼中的 TTX。我们使用捕获-SELEX 结合下一代测序技术，鉴定出适配体，并评估了它们的结合特性。最后，开发了一种高度灵敏且易于使用的杂交抗体-适配体夹心检测法，与文献中报道的几种检测法和商业免疫试剂盒相比，该方法具有更优的性能。该检测法成功地应用于河豚鱼提取物中 TTX 的定量检测，所得结果与平行进行的基于竞争磁珠的免疫检测法非常吻合。这是文献中报道的为数不多的小分子分析物的此类杂交检测法之一，并且还证明了其与现场样本的兼容性。
