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针对食源性病原体细菌的适配体的筛选。

Selection of aptamers targeted to food-borne pathogenic bacteria .

作者信息

Wang Lan, Lyu Shuxia, Gu Ganyu, Bolten Samantha

机构信息

College of Food Science and Engineering Shenyang Agricultural University Shenyang China.

College of Bioscience and Biotechnology Shenyang Agricultural University Shenyang China.

出版信息

Food Sci Nutr. 2020 Jun 3;8(7):3835-3842. doi: 10.1002/fsn3.1677. eCollection 2020 Jul.

DOI:10.1002/fsn3.1677
PMID:32724645
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7382169/
Abstract

() is a common marine halophilic food-borne pathogen, mainly found in seafood and food with a high salt content. Gastrointestinal reactions such as diarrhea, headache, vomiting, nausea, and abdominal cramps may occur after eating food infected with . This study aimed to screen for high-affinity aptamers that specifically recognize . A high-affinity modified aptamer screening kit was used to rapidly screen aptamers of the food-borne . The first round of screening involved release of target aptamers from the microspheres. The "false-positive" aptamers were eliminated after specific binding to and elution of in the second round. The second round of screening of the aptamers involved polymerase chain reaction (PCR), and the abundance of a sequence was determined using next-generation sequencing. Nine high-affinity aptamer sequences were obtained, and the first eight modified aptamer sequences were derived using a cloud-based intelligent software of the American AM Biotech Co. was used as a control, and aptamer ID 12 with the highest affinity for was selected using real-time PCR. According to the principle of color change caused by nano-gold condensing under salt induction, , , and were used as counter-screening bacteria, and the aptamer ID12 was combined with nano-gold. The results showed that aptamer ID12 has strong specificity for . Based on these findings, this study developed a simple, innovative, and rapid method for screening aptamers.

摘要

()是一种常见的海洋嗜盐食源性病原体,主要存在于海鲜和高盐含量的食物中。食用感染了()的食物后可能会出现腹泻、头痛、呕吐、恶心和腹部绞痛等胃肠道反应。本研究旨在筛选特异性识别()的高亲和力适配体。使用高亲和力修饰适配体筛选试剂盒快速筛选食源性病原体()的适配体。第一轮筛选涉及从微球中释放目标适配体。在第二轮中,通过与()特异性结合并洗脱后,消除“假阳性”适配体。第二轮适配体筛选涉及聚合酶链反应(PCR),并使用下一代测序确定序列的丰度。获得了9个高亲和力适配体序列,前8个修饰适配体序列是使用美国AM生物技术公司基于云的智能软件推导出来的。()用作对照,并使用实时PCR选择对()亲和力最高的适配体ID 12。根据盐诱导下纳米金凝聚引起颜色变化的原理,使用()、()和()作为反筛选细菌,并将适配体ID12与纳米金结合。结果表明,适配体ID12对()具有很强的特异性。基于这些发现,本研究开发了一种简单、创新且快速的筛选()适配体的方法。

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