| Establishment of a viable cell detection system for microorganisms in wine based on ethidium monoazide and quantitative PCR | |
Shi, Hui1; Xu, Wentao1; Quoclinh Trinh1; Luo, Yunbo1,2; Liang, Zhihong2; Li, Yingcong3 ; Huang, Kunlun1,2; Xu, WT (reprint author), China Agr Univ, Coll Food Sci & Nutr Engn, Lab Food Safety, Beijing 100083, Peoples R China.
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| 心理所单位排序 | 3 |
| 摘要 | Fermentability and contamination level of wine can be assessed through the detection of viable fermentation-related and spoilage-related microorganisms. Ethidium monoazide in combination with quantitative PCR (EMA-qPCR) has been considered as a promising method to enumerate viable cells. Milling for 80 s by O 500-mu m glass beads is demonstrated to be optimal for DNA extraction from yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) in wine to be used as a template for PCR. EMA-qPCR results from experiments using DNA extracted by this method correlate well with the results of a plating assay (R-2 > 0.99), and a PCR efficiency between 96% and 105% was obtained. Moreover, for all of these microorganisms, EMA treatment of pure cultures at a low concentration (10 mu g/mL) for 20 min photoactivation resulted in effective differentiation between viable and non-viable cells and had no effect on viable cells. Due to sublethal injury to some cells, underestimation of cell counts was found in most of the wine samples tested using the EMA-qPCR method, and a 40-min incubation in recovery medium could completely offset this error. Our results suggest an optimal glass-bead DNA extraction method and EMA treatment suitable for all of the main microorganisms in wine. The EMA-qPCR method was successfully applied to quantify yeasts. Saccharomyces cerevisiae (S. cerevisiae), LAB, non-Oenococcus oeni LAB (non-O. oeni LAB) and AAB in wine samples. (C) 2012 Elsevier Ltd. All rights reserved. |
| 关键词 | Cell viability Wine Ethidium monoazide Quantitative PCR |
| 学科领域 | Physiological Psychology/biological Psychology |
| 2012-09-01 | |
| 语种 | 英语 |
| 发表期刊 | FOOD CONTROL
 |
| ISSN | 0956-7135 |
| 卷号 | 27期号:1页码:81-86 |
| 期刊论文类型 | Article |
| URL | 查看原文 |
| 收录类别 | SCI |
| 项目简介 | This work was supported by the Ministry of Science and Technology of China (2011BAK10B02). |
| WOS记录号 | WOS:000304790800012 |
| 资助机构 | Ministry of Science and Technology of China [2011BAK10B02] |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.psych.ac.cn/handle/311026/11500 |
| 专题 | 管理支撑系统 |
| 通讯作者 | Xu, WT (reprint author), China Agr Univ, Coll Food Sci & Nutr Engn, Lab Food Safety, Beijing 100083, Peoples R China. |
| 作者单位 | 1.China Agr Univ, Coll Food Sci & Nutr Engn, Lab Food Safety, Beijing 100083, Peoples R China 2.Minist Agr, Supervis & Testing Ctr Agr Prod Qual, Beijing 100083, Peoples R China 3.Chinese Acad Sci, Inst Psychol, Beijing 100101, Peoples R China |
| 推荐引用方式 GB/T 7714 | Shi, Hui,Xu, Wentao,Quoclinh Trinh,et al. Establishment of a viable cell detection system for microorganisms in wine based on ethidium monoazide and quantitative PCR[J]. FOOD CONTROL,2012,27(1):81-86. |
| APA | Shi, Hui.,Xu, Wentao.,Quoclinh Trinh.,Luo, Yunbo.,Liang, Zhihong.,...&Xu, WT .(2012).Establishment of a viable cell detection system for microorganisms in wine based on ethidium monoazide and quantitative PCR.FOOD CONTROL,27(1),81-86. |
| MLA | Shi, Hui,et al."Establishment of a viable cell detection system for microorganisms in wine based on ethidium monoazide and quantitative PCR".FOOD CONTROL 27.1(2012):81-86. |
| 条目包含的文件 | ||||||
| 文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
| WOS_000304790800012.（369KB） | 开放获取 | Apache License | 浏览 请求全文 | |||
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