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液相色谱-紫外检测及对在不同条件下培养的产肠毒素大肠杆菌所产生的热不稳定毒素进行定量分析。

Liquid chromatography-ultraviolet detection and quantification of heat-labile toxin produced by enterotoxigenic E. coli cultured under different conditions.

作者信息

Sospedra Isabel, De Simone Carmela, Soriano José Miguel, Mañes Jordi, Ferranti Pasquale, Ritieni Alberto

机构信息

Department of Nursing, Faculty of Health Science, University of Alicante, Campus Sant Vicent del Raspeig, Ap. 99, E-03080, Alicante, Spain; Research Group on Food and Nutrition (ALINUT), Spain.

Department of Agriculture, University of Naples Federico II, Parco Gussone, Portici, I-80055, Italy; Institute of Food Science and Technology, National Research Council, Via Roma 64a-c, Avellino I-83100, Italy.

出版信息

Toxicon. 2018 Jan;141:73-78. doi: 10.1016/j.toxicon.2017.11.014. Epub 2017 Dec 2.

Abstract

Enterotoxigenic Escherichia coli (ETEC) is the main bacterial cause of dehydrating infant diarrhoea in less-developed countries. Labile toxin (LT) is the major virulent factor of ETEC. Easy diagnostic tests are necessary to reduce the number of cases. Immunological methods have some drawbacks and also have important limitations. For that reason, a Liquid Chromatography coupled to UV detector technique (LC-UV) has been optimize to a rapid identification and quantification of LT from bacteria cultures. It is also important to know optimal conditions for LT and with this purpose several enterotoxigenic E. coli strains have been studied to determine the influence of glucose concentration and different culture media on LT production. LC-UV technique demonstrated to be a good method for LT quantification showing values of 15 ng/mL and 45 ng/mL for limits of detection and quantification respectively. LT quantification revealed that toxin production is directly related to the concentration of glucose added in the broth medium. Tryptic Soy Broth is the most efficient culture medium for E. coli growth and enterotoxin production.

摘要

产肠毒素大肠杆菌(ETEC)是欠发达国家婴儿腹泻脱水的主要细菌病因。不耐热毒素(LT)是ETEC的主要致病因素。需要简便的诊断测试来减少病例数量。免疫方法存在一些缺点,也有重要局限性。因此,液相色谱联用紫外检测器技术(LC-UV)已得到优化,用于从细菌培养物中快速鉴定和定量LT。了解LT的最佳条件也很重要,为此研究了几种产肠毒素大肠杆菌菌株,以确定葡萄糖浓度和不同培养基对LT产生的影响。LC-UV技术被证明是LT定量的良好方法,检测限和定量限分别为15 ng/mL和45 ng/mL。LT定量显示,毒素产生与肉汤培养基中添加的葡萄糖浓度直接相关。胰蛋白胨大豆肉汤是大肠杆菌生长和产肠毒素最有效的培养基。

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