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Multiplex biotoxin surface plasmon resonance method for marine biotoxins in algal and seawater samples

机译:藻类和海水样品中海洋生物毒素的多重生物毒素表面等离子体共振方法

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A multiplex surface plasmon resonance (SPR) biosensor method for the detection of paralytic shellfish poisoning (PSP) toxins, okadaic acid (and analogues) and domoic acid was developed. This method was compared to enzyme-linked immunosorbent assay (ELISA) methods. Seawater samples (n=256) from around Europe were collected by the consortia of an EU project MIcroarrays for the Detection of Toxic Algae (MIDTAL) and evaluated using each method. A simple sample preparation procedure was developed which involved lysing and releasing the toxins from the algal cells with glass beads followed by centrifu-gation and filtering the extract before testing for marine biotoxins by both multi-SPR and ELISA. Method detection limits based on IC_(20) values for PSP, okadaic acid and domoic acid toxins were 0.82, 0.36 and 1.66 ng/ml, respectively, for the prototype multiplex SPR biosensor. Evaluation by SPR for seawater samples has shown that 47, 59 and 61 % of total seawater samples tested positive (result greater than the IC_(20)) for PSP, okadaic acid (and analogues) and domoic acid toxins, respectively. Toxic samples were received mainly from Spain and Ireland. This work has demonstrated the potential of multiplex analysis for marine biotoxins in algal and seawater samples with results available for 24 samples within a 7 h period for three groups of key marine biotoxins. Multiplex immunologicai methods could therefore be used as early warning monitoring tools for a variety of marine biotoxins in seawater samples.
机译:建立了一种用于检测麻痹性贝类中毒(PSP)毒素,冈田酸(及其类似物)和多聚酸的多重表面等离子体共振(SPR)生物传感器方法。将该方法与酶联免疫吸附测定(ELISA)方法进行了比较。来自欧洲各地的海水样品(n = 256)由欧盟项目有毒藻类检测微阵列(MIDTAL)的财团收集,并使用每种方法进行评估。开发了一种简单的样品制备程序,包括用玻璃珠将藻类细胞中的毒素裂解并释放,然后离心并过滤提取物,然后通过多重SPR和ELISA测试海洋生物毒素。对于原型多重SPR生物传感器,基于IC_(20)值的PSP,冈田酸和十二酸毒素的方法检测限分别为0.82、0.36和1.66 ng / ml。通过SPR对海水样品进行的评估表明,分别占总海水样品47%,59%和61%的PSP,冈田酸(和类似物)和海藻酸毒素呈阳性(结果大于IC_(20))。有毒样品主要来自西班牙和爱尔兰。这项工作证明了藻类和海水样品中海洋生物毒素的多重分析的潜力,三组关键海洋生物毒素在7小时内可获得24个样品的结果。因此,多重免疫学方法可以用作海水样本中各种海洋生物毒素的预警监测工具。

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