Quantitative real-time PCR method for rapid enumeration of Enterobacteriaceae in food

食品中肠杆菌科细菌快速计数的实时荧光定量PCR方法

• 批准号: 22780191
• 负责人: TAKAHASHI Hajime
• 金额: $ 2.5万
• 依托单位: Tokyo University of Marine Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22780191/
• 关键词: 食品衛生; 食品微生物; 遺伝子; 腸内細菌科菌群; リアルタイムPCR

In food manufacturing companies, the bacteriological examination of foods targets not only foodborne pathogens but total aerobic bacterial counts and hygiene indicator bacteria as well. However, it is hard to establish an examination method using real-time PCR for total aerobic bacteria and hygiene indicator bacteria, since the method needs to target several different bacterial species at the same time. In this study, we developed a rapid detection and quantification method for Enterobacteriaceae using quantitative real-time PCR.First of all, several DNA regions were investigated in order to design PCR primers for specific detection of Enterobacteriaceae. As a result, primers designed within the rplP gene could specifically detect Enterobacteriaceae.Next, the accuracy of the real-time PCR method was confirmed by comparing with a culture method. The difference between these two methods was within one order. These results indicate that the real-timePCR developed in this study could accurately and rapidly quantify Enterobacteriaceae in foods.

在食品生产企业中，食品的细菌学检查不仅针对食源性病原体，还针对需氧菌总数和卫生指示菌。然而，很难建立使用实时PCR检测总需氧菌和卫生指示菌的方法，因为该方法需要同时针对几种不同的细菌物种。本研究建立了一种肠杆菌科细菌的实时荧光定量PCR快速检测方法。结果，在rplP基因内设计的引物可以特异性地检测肠杆菌科。接下来，通过与培养方法进行比较，证实了实时PCR方法的准确性。这两种方法之间的差距在一个数量级之内。本研究建立的实时荧光定量PCR方法能够准确、快速地定量食品中的肠杆菌。

项目成果

PCR法を用いた腸内細菌科菌群の迅速検出法の検討

PCR法快速检测肠杆菌科细菌的方法研究

• 发表时间: 2011
• 作者: 斉藤瑠美,田中悠一郎;高橋 肇,久田 孝,木村 凡
• 通讯作者: 高橋 肇,久田 孝,木村 凡

Bon Kimura Quantitative real-time PCR method for rapid enumeration of Enterobacteriaceae in food

Bon Kimura 食品中肠杆菌科细菌快速计数的实时荧光定量PCR方法

• 发表时间: 2012
• 作者: Hajime Takahashi;Yuichiro Tanaka;Rumi Sito;Takashi Kuda
• 通讯作者: Takashi Kuda

Quantitative real-time PCR method for rapid enumeration of Enterobacteriaceae in food

食品中肠杆菌科细菌快速计数的实时荧光定量PCR方法

• 发表时间: 2012
• 作者: Hajime Takahashi;Yuichiro Tanaka;Rumi Sito;Takashi Kuda;水野伸也;Hajime Takahashi
• 通讯作者: Hajime Takahashi

腸内細菌科菌群検出方法

肠杆菌科菌群检测方法

• 发表时间: 2012