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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.
2012-09-01
发表期刊FOOD CONTROL
ISSN0956-7135
文章类型Article
卷号27期号:1页码:81-86
产权排序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
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收录类别SCI
语种英语
项目资助者Ministry of Science and Technology of China [2011BAK10B02]
项目简介This work was supported by the Ministry of Science and Technology of China (2011BAK10B02).
WOS记录号WOS:000304790800012
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文献类型期刊论文
条目标识符http://ir.psych.ac.cn/handle/311026/11500
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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
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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.
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