Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid
Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid
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用于野百合碱（一种有毒吡咯里西啶生物碱）灵敏免疫色谱分析的预孵育形式
DOI：
10.1002/pca.2838
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发表时间：
2019
影响因子：
3.3
通讯作者：
S.
中科院分区：
文献类型：
--
作者：
Yusakul;G.;Sakamoto;S.;Chanpokapaiboon;K.;Tanaka;H.;Morimoto;S.
MCT （monocrotaline）被归类为1,2 -脱氢吡咯利西啶生物碱（DHPA），是一种主要由crotalariaceae产生的有毒化合物。谷物和草药中的MCT污染会导致肝炎、肠胃炎、肺血管炎和高血压，以及不同类型的癌症。目前MCT的分析方法复杂且昂贵，采用液相色谱和质谱相结合的方法。目的：建立一种简便、灵敏的免疫层析（PI‐ICA）检测MCT的预孵育方法。方法：在试纸浸渍前，将含有MCT的样品与胶体金偶联的抗MCT单克隆抗体孵育。我们比较了PI‐ICA与传统ICA （Conv‐ICA）格式的检测灵敏度。结果PI‐ICA的检测限（LOD）为0.61 ng/mL，是Conv‐ICA格式的16倍。这些结果表明，PI - ICA方法对MCT具有高的结合特异性，对逆转录酶、逆转录酶、顺转录酶和顺转录酶具有低的交叉反应性。通过PI‐ICA分析，含有MCT和相关DHPAs的植物样品溶液产生了阳性结果。结论与Conv‐ICA相比，PI‐ICA系统具有较高的灵敏度。此外，所开发的PI‐ICA方法简单有效，可用于检测MCT污染。
IntroductionMonocrotaline (MCT), which is classified as a 1,2‐dehydropyrrolizidine alkaloid (DHPA), is a toxic compound that is mainly produced byCrotalariaspp. MCT contamination in cereals and herbs leads to hepatitis, gastroenteritis, pulmonary vasculitis and hypertension, and different types of cancer. The current analytical methods for MCT are complicated and expensive using liquid chromatography equipped with mass spectrometry detection.ObjectiveThe aim of this study was to develop a simple and sensitive preincubation format for an immunochromatographic assay (PI‐ICA) for MCT detection.MethodologyWe conducted the PI‐ICA via incubation of an MCT‐containing sample with an anti‐MCT monoclonal antibody conjugated with colloidal gold before strip dipping. We compared the PI‐ICA detection sensitivity with that of the conventional ICA (Conv‐ICA) format.ResultsThe PI‐ICA was sensitive with a limit of detection (LOD) of 0.61 ng/mL, which is a 16‐fold improvement over the Conv‐ICA format. These results indicated that the PI‐ICA method exhibits high binding specificity for MCT and low cross‐reactivity towards retronecine, retrorsine, senecionine and heliotrine. Sample solutions from plants containing MCT and related DHPAs produced positive results via PI‐ICA analysis.ConclusionsThe proposed PI‐ICA system provides a highly sensitive method compared to Conv‐ICA. In addition, the developed PI‐ICA method is simple and highly effective for detecting MCT contamination.