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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
Source PublicationFOOD CONTROL
ISSN0956-7135
SubtypeArticle
Volume27Issue:1Pages:81-86
Contribution Rank3
AbstractFermentability 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.
KeywordCell viability Wine Ethidium monoazide Quantitative PCR
Subject AreaPhysiological Psychology/biological Psychology
URL查看原文
Indexed BySCI
Language英语
Funding OrganizationMinistry of Science and Technology of China [2011BAK10B02]
Project Intro.This work was supported by the Ministry of Science and Technology of China (2011BAK10B02).
WOS IDWOS:000304790800012
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Cited Times:15[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.psych.ac.cn/handle/311026/11500
Collection支撑系统
Corresponding AuthorXu, WT (reprint author), China Agr Univ, Coll Food Sci & Nutr Engn, Lab Food Safety, Beijing 100083, Peoples R China.
Affiliation1.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
Recommended Citation
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.
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