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用于从增菌培养物中检测沙门氏菌属的蛋白质芯片

Protein Chips for Detection of Salmonella spp. from Enrichment Culture.

作者信息

Poltronieri Palmiro, Cimaglia Fabio, De Lorenzis Enrico, Chiesa Maurizio, Mezzolla Valeria, Reca Ida Barbara

机构信息

CNR-ISPA, Institute of Sciences of Food Productions, via Monteroni km 7, 73100 Lecce, Italy.

Biotecgen, c/o Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, Italy.

出版信息

Sensors (Basel). 2016 Apr 22;16(4):574. doi: 10.3390/s16040574.

Abstract

Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions.

摘要

食源性病原体是食源性疾病流行的原因,因此有必要在食品生产过程中快速检测这些病原体。先进行预富集培养,然后进行选择性琼脂平板培养是标准的检测方法。诸如qPCR之类的分子方法已经提供了一种在富集培养24小时内检测病原体的快速方案。生物传感器也可能提供一种快速工具,以便在预富集培养的早期确定沙门氏菌污染的来源。按照法国标准化协会验证的qPCR方案,使用Pathatrix通过免疫分离处理40 mL沙门氏菌属富集培养物。结合免疫分离的沙门氏菌生物传感器检测限为100个细菌/40 mL,比之前的结果提高了400倍。qPCR分析需要用裂解缓冲液处理磁珠结合的细菌并进行DNA纯化,检测限为2 cfu/50 μL。最后,开发了一种蛋白质芯片,并用于筛选和鉴定5种常见病原体,即沙门氏菌属、大肠杆菌、金黄色葡萄球菌、弯曲杆菌属和李斯特菌属。该蛋白质芯片在物种鉴定方面具有高特异性,建议将其集成到芯片实验室系统中,用于快速且可重复地筛选污染食品生产的沙门氏菌属和其他病原体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4893/4851088/6819cdc8b11f/sensors-16-00574-g001.jpg

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