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用于检测受污染食品中16种细菌病原体的多重16S rRNA衍生基因生物芯片。

Multiplex 16S rRNA-derived geno-biochip for detection of 16 bacterial pathogens from contaminated foods.

作者信息

Shin Hwa Hui, Hwang Byeong Hee, Cha Hyung Joon

机构信息

Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea.

Division of Bioengineering, Incheon National University, Incheon, Korea.

出版信息

Biotechnol J. 2016 Nov;11(11):1405-1414. doi: 10.1002/biot.201600043. Epub 2016 Sep 6.

Abstract

Foodborne diseases caused by various pathogenic bacteria occur worldwide. To prevent foodborne diseases and minimize their impacts, it is important to inspect contaminated foods and specifically detect many types of pathogenic bacteria. Several DNA oligonucleotide biochips based on 16S rRNA have been investigated to detect bacteria; however, a mode of detection that can be used to detect diverse pathogenic strains and to examine the safety of food matrixes is still needed. In the present work, a 16S rRNA gene-derived geno-biochip detection system was developed after screening DNA oligonucleotide specific capture probes, and it was validated for multiple detection of 16 pathogenic strains that frequently occur with a signature pattern. rRNAs were also used as detection targets directly obtained from cell lysates without any purification and amplification steps in the bacterial cells separated from 8 food matrixes by simple pretreatments. Thus, the developed 16S rRNA-derived geno-biochip can be successfully used for the rapid and multiple detection of the 16 pathogenic bacteria frequently isolated from contaminated foods that are important for food safety.

摘要

由各种致病细菌引起的食源性疾病在全球范围内都有发生。为预防食源性疾病并将其影响降至最低,检查受污染食品并特异性检测多种致病细菌非常重要。已经研究了几种基于16S rRNA的DNA寡核苷酸生物芯片来检测细菌;然而,仍然需要一种可用于检测多种致病菌株并检查食品基质安全性的检测模式。在本研究中,在筛选DNA寡核苷酸特异性捕获探针后,开发了一种基于16S rRNA基因的基因生物芯片检测系统,并对16种经常以特征模式出现的致病菌株进行了多重检测验证。rRNAs还被用作直接从细胞裂解物中获得的检测靶标,在通过简单预处理从8种食品基质中分离出的细菌细胞中,无需任何纯化和扩增步骤。因此,所开发的基于16S rRNA的基因生物芯片可成功用于快速、多重检测从受污染食品中频繁分离出的16种对食品安全至关重要的致病细菌。

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