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利用培养的成纤维细胞中的蛋白质合成抑制作用快速灵敏地筛查玉米中呕吐毒素的方法

Fast and sensitive screening method for detection of trichothecenes in maize by using protein synthesis inhibition in cultured fibroblasts.

作者信息

Scossa-Romano D A, Bickel R E, Zweifel U, Reinhardt C A, Lüthy J W, Schlatter C L

出版信息

J Assoc Off Anal Chem. 1987 Jan-Feb;70(1):129-32.

PMID:3558266
Abstract

A fast and sensitive bioassay with hamster (BHK-21 C13) fibroblasts for the detection of toxic trichothecenes in maize is described. Cells are exposed to pure toxins or crude maize extracts for 30 min. The mixture is then incubated with [1-14C]-leucine for an additional 60 min and the radioactivity incorporated into the protein of the washed cells is determined. The sensitivity of the assay was in the range 1-10 ng/mL (or 50 ppb in maize) for T-2, HT-2, and diacetoxyscirpenol. At least 1000-fold higher concentrations of non-trichothecene mycotoxins and plant toxins were necessary to cause an inhibition of protein synthesis in the cells. Of 24 maize samples tested, 14 gave a positive response in this assay and the presence of trichothecenes could be confirmed chemically in 11 samples. Therefore, the described bioassay is proposed as a useful screening method for cytotoxic trichothecenes in maize.

摘要

本文描述了一种利用仓鼠(BHK-21 C13)成纤维细胞快速灵敏地检测玉米中有毒单端孢霉烯族毒素的生物测定方法。将细胞暴露于纯毒素或玉米粗提物中30分钟。然后将混合物与[1-¹⁴C]-亮氨酸一起孵育60分钟,并测定掺入洗涤后细胞蛋白质中的放射性。该测定方法对T-2毒素、HT-2毒素和二醋酸雪腐镰刀菌烯醇的灵敏度范围为1-10纳克/毫升(或玉米中50 ppb)。非单端孢霉烯族霉菌毒素和植物毒素的浓度至少要高1000倍才能抑制细胞中的蛋白质合成。在测试的24个玉米样品中,有14个在该测定中呈阳性反应,并且在11个样品中通过化学方法确认了单端孢霉烯族毒素的存在。因此,所描述的生物测定方法被提议作为检测玉米中具有细胞毒性的单端孢霉烯族毒素的一种有用的筛选方法。

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