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富集培养基对生菜和番茄富集培养物中肠炎沙门氏菌实时检测的影响。

Effect of enrichment medium on real-time detection of Salmonella enterica from lettuce and tomato enrichment cultures.

机构信息

U.S. Department of Agriculture, Agricultural Research Service, Produce Safety and Microbiology Research Unit, Albany, California 94710, USA.

出版信息

J Food Prot. 2010 Jun;73(6):1047-56. doi: 10.4315/0362-028x-73.6.1047.

Abstract

Three enrichment broths commonly used for detection of Salmonella (buffered peptone water [BPW], tryptic soy broth [TSB], and universal preenrichment broth [UPB]) were compared for use in real-time SYBR Green PCR detection of Salmonella introduced into enrichment cultures made from store-bought lettuce and tomatoes. The produce served as a source of normal plant microbiota to measure how well DNA-based detection methods for Salmonella work in a suspension of plant-associated bacteria that may be closely related to Salmonella. A qualitative assessment of the background microbiota that grew in the three enrichment broths cultures from tomato and lettuce samples revealed that different bacteria predominated in the different broths. Results obtained with five produce-related outbreak Salmonella strains and PCR primers directed toward three different Salmonella genes suggest that the ability to detect Salmonella from these enrichment cultures by real-time PCR was 10 to 1,000 times better with TSB enrichment cultures. Detection levels were similar between the different enrichment media when an immunomagnetic separation method was used; however, the immunological technique did not enhance detection from TSB enrichment cultures. Detection could be affected by the medium and by the background microbiota. An immunomagnetic separation method may be useful in BPW and UPB enrichment cultures but not in TSB enrichment cultures.

摘要

三种常用的富集培养基(缓冲蛋白胨水[BPW]、胰蛋白酶大豆肉汤[TSB]和通用预富集肉汤[UPB])用于实时 SYBR Green PCR 检测从市售生菜和番茄中引入的富集培养物中的沙门氏菌。这些农产品作为正常植物微生物群的来源,用于衡量基于 DNA 的沙门氏菌检测方法在与沙门氏菌密切相关的植物相关细菌悬浮液中的工作效果。对来自番茄和生菜样本的三种富集培养基培养物中生长的背景微生物群的定性评估表明,不同的细菌在不同的培养基中占优势。针对三种不同沙门氏菌基因的五个与农产品相关的暴发沙门氏菌菌株和 PCR 引物的结果表明,通过实时 PCR 从这些富集培养物中检测沙门氏菌的能力在 TSB 富集培养物中提高了 10 到 1000 倍。当使用免疫磁分离方法时,不同的富集培养基之间的检测水平相似;然而,免疫技术并没有增强 TSB 富集培养物的检测效果。检测可能受到培养基和背景微生物群的影响。免疫磁分离方法在 BPW 和 UPB 富集培养物中可能有用,但在 TSB 富集培养物中则不然。

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