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利用实时聚合酶链反应结合免疫磁珠分离和全基因组扩增技术快速检测生鸡胸肉中的沙门氏菌。

Rapid detection of Salmonella in raw chicken breast using real-time PCR combined with immunomagnetic separation and whole genome amplification.

作者信息

Hyeon Ji-Yeon, Deng Xiangyu

机构信息

Center for Food Safety, Department of Food Science and Technology, University of Georgia, 1109 Experiment St, Griffin, GA, 30223, USA.

Center for Food Safety, Department of Food Science and Technology, University of Georgia, 1109 Experiment St, Griffin, GA, 30223, USA.

出版信息

Food Microbiol. 2017 May;63:111-116. doi: 10.1016/j.fm.2016.11.007. Epub 2016 Nov 21.

Abstract

We presented the first attempt to combine immunomagnetic separation (IMS), whole genome amplification by multiple displacement amplification (MDA) and real-time PCR for detecting a bacterial pathogen in a food sample. This method was effective in enabling real-time PCR detection of low levels of Salmonella enterica Serotype Enteritidis (SE) (∼10 CFU/g) in raw chicken breast without culture enrichment. In addition, it was able to detect refrigeration-stressed SE cells at lower concentrations (∼0.1 CFU/g) in raw chicken breast after a 4-h culture enrichment, shortening the detection process from days to hours and displaying no statistical difference in detection rate in comparison with a culture-based detection method. By substantially improving performance in SE detection over conventional real-time PCR, we demonstrated the potential of IMS-MDA real-time PCR as a rapid, sensitive and affordable method for detecting Salmonella in food.

摘要

我们首次尝试将免疫磁珠分离(IMS)、多重置换扩增(MDA)全基因组扩增和实时荧光定量PCR相结合,用于检测食品样本中的细菌病原体。该方法可有效实现对未经过培养富集的生鸡胸肉中低水平肠炎沙门氏菌(SE)(约10 CFU/g)的实时荧光定量PCR检测。此外,经过4小时的培养富集后,该方法能够检测出生鸡胸肉中浓度更低(约0.1 CFU/g)的冷藏应激SE菌,将检测过程从数天缩短至数小时,并且与基于培养的检测方法相比,检测率无统计学差异。通过大幅提高与传统实时荧光定量PCR相比的SE检测性能,我们证明了IMS-MDA实时荧光定量PCR作为一种快速、灵敏且经济实惠的食品中沙门氏菌检测方法的潜力。

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