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基于TaqMan的一步法逆转录实时PCR检测鲜切蔬菜中的活肠炎沙门氏菌。

Detection of live Salmonella enterica in fresh-cut vegetables by a TaqMan-based one-step reverse transcription real-time PCR.

作者信息

Miao Y J, Xiong G T, Bai M Y, Ge Y, Wu Z F

机构信息

Department of Food Science and Engineering, School of Marine Science, Ningbo University, Ningbo, China.

Food Science Institute, Zhejiang Academy of Agricultural Science, Hangzhou, China.

出版信息

Lett Appl Microbiol. 2018 May;66(5):447-454. doi: 10.1111/lam.12871. Epub 2018 Mar 24.

Abstract

UNLABELLED

Fresh-cut produce is at greater risk of Salmonella contamination. Detection and early warning systems play an important role in reducing the dissemination of contaminated products. One-step Reverse Transcription Polymerase Chain Reaction (RT-qPCR) targeting Salmonella tmRNA with or without a 6-h enrichment was evaluated for the detection of Salmonella in fresh-cut vegetables after 6-h storage. LOD of one-step RT-qPCR was 1·0 CFU per ml (about 100 copies tmRNA per ml) by assessed 10-fold serially diluted RNA from 10 CFU per ml bacteria culture. Then, one-step RT-qPCR assay was applied to detect viable Salmonella cells in 14 fresh-cut vegetables after 6-h storage. Without enrichment, this assay could detect 10 CFU per g for fresh-cut lettuce, cilantro, spinach, cabbage, Chinese cabbage and bell pepper, and 10 CFU per g for other vegetables. With a 6-h enrichment, this assay could detect 10 CFU per g for all fresh-cut vegetables used in this study. Moreover, this assay was able to discriminate viable cells from dead cells. This rapid detection assay may provide potential processing control and early warning method in fresh-cut vegetable processing to strengthen food safety assurance.

SIGNIFICANCE AND IMPACT OF THE STUDY

Significance and Impact of the Study: Fresh-cut produce is at greater risk of Salmonella contamination. Rapid detection methods play an important role in reducing the dissemination of contaminated products. One-step RT-qPCR assay used in this study could detect 10 CFU per g Salmonella for 14 fresh-cut vegetables with a 6-h short enrichment. Moreover, this assay was able to discriminate viable cells from dead cells. This rapid detection assay may provide potential processing control and early warning method in fresh-cut vegetable processing to strengthen food safety assurance.

摘要

未标记

鲜切农产品受沙门氏菌污染的风险更高。检测和预警系统在减少受污染产品的传播方面发挥着重要作用。评估了针对沙门氏菌tmRNA的一步法逆转录聚合酶链反应(RT-qPCR),在6小时储存后,对鲜切蔬菜中的沙门氏菌进行检测,该方法有或没有6小时增菌步骤。通过对每毫升10 CFU细菌培养物中10倍系列稀释的RNA进行评估,一步法RT-qPCR的检测限为每毫升1.0 CFU(约每毫升100拷贝tmRNA)。然后,将一步法RT-qPCR检测应用于检测6小时储存后的14种鲜切蔬菜中的活沙门氏菌细胞。不进行增菌时,该检测方法可检测出每克鲜切生菜、香菜、菠菜、卷心菜、大白菜和甜椒中含有10 CFU,其他蔬菜中每克含有10 CFU。经过6小时增菌后,该检测方法可检测出本研究中所有鲜切蔬菜中每克含有10 CFU。此外,该检测方法能够区分活细胞和死细胞。这种快速检测方法可能为鲜切蔬菜加工提供潜在的加工控制和预警方法,以加强食品安全保障。

研究的意义和影响

鲜切农产品受沙门氏菌污染的风险更高。快速检测方法在减少受污染产品的传播方面发挥着重要作用。本研究中使用的一步法RT-qPCR检测方法在经过6小时短时间增菌后,可检测出14种鲜切蔬菜中每克含有10 CFU的沙门氏菌。此外,该检测方法能够区分活细胞和死细胞。这种快速检测方法可能为鲜切蔬菜加工提供潜在的加工控制和预警方法,以加强食品安全保障。

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