Improved detection and enumeration of yeasts in wine by Cells-qPCR

Soares-Santos, Verónica; Pardo, Isabel; Ferrer, Sergi

doi:10.1016/j.lwt.2017.12.007

Cited by 9 publications

(6 citation statements)

References 27 publications

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“…For the extracted DNA, the sample volume used in each reaction was 2.5 μL as indicated by Hayashi et al (2007) . Nevertheless, regarding the whole cells, fivefold higher sample volumes were applied to each LAMP reaction to enhance the method’s sensitivity ( Soares-Santos et al, 2018 ). For both DNA and cells, the results showed the presence of turbidity by directly observing the reaction tubes ( Figure 1 ).…”

Section: Resultsmentioning

confidence: 99%

“…The obtained LoDs were 10 3 cells/mL (12 cells/reaction tube) in white wine, and 10 4 cells/mL (125 cells/reaction tube) in red wine (data not shown). To lower the O. oeni LoD in both matrices, the effect of a previous cell wall mechanical lysis was evaluated ( Soares-Santos et al, 2018 ). To this end, the assay was repeated, but before the Cells-LAMP amplification, all the cell suspensions were subjected to a mechanical lysis.…”

Section: Resultsmentioning

confidence: 99%

“…were grown in MRS (Scharlau, Barcelona, Spain) supplemented with 0.5 g/L cysteine (Merck, Darmstadt, Germany). Yeasts were grown in YPD ( Soares-Santos et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

“…After washing the cell suspensions, each dilution was used as a template for the LAMP assay. Besides whole cells, the mechanically lysed cells were also assayed ( Soares-Santos et al, 2018 ). For this purpose, after washing cells 425–600 μm-diameter glass beads (Sigma, St. Louis, United States) were added to each cell suspension [50% (w/v)], and each dilution was washed and lysed separately.…”

Section: Methodsmentioning

confidence: 99%

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Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP

2018

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Fast detection and enumeration of Oenococcus oeni in winemaking are necessary to determine whether malolactic fermentation (MLF) is likely to be performed or not and to decide if the use of a commercial starter is needed. In other wines, however, performing MLF can be detrimental for wine and should be avoided. The traditional identification and quantification of this bacteria using culture-dependent techniques in wine-related matrices require up to 14 days to yield results, which can be a very long time to perform possible enological operations. Loop-mediated isothermal amplification (LAMP) is a novel culture-independent technique that amplifies nucleic acid sequences under isothermal conditions with high specificity and efficiency in less than 1 h with inexpensive equipment. We designed LAMP primers for the specific detection and quantification of O. oeni cells. The developed LAMP method allows O. oeni to be detected directly from both grape musts and wines within 1 h from the time that the LAMP reaction begins, and without DNA extraction and purification requirements. The high sensitivity of LAMP methodology is achieved by previous mechanical cells lysis with no further purification by detecting one single cell per reaction in culture media, and in white/red grape musts and wines by avoiding reaction inhibition by ethanol, polyphenols, and other wine inhibitors. Cells can be concentrated prior to the LAMP reaction to further increase this sensitivity. Moreover, the LAMP method does not require expensive equipment and can be easily operated. The developed method is both economic and fast and offers high sensitivity and specificity.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…were grown in MRS (Scharlau, Barcelona, Spain) supplemented with 0.5 g/L cysteine (Merck, Darmstadt, Germany). Yeasts were grown in YPD ( Soares-Santos et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP

2018

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“…However, this method relied on large instruments and skilled operators. Molecular techniques, such as polymerase chain reaction [9,10], have high sensitivity and specificity but are expensive [11]. Biofunctionalized resonant photonic grating sensors [12] were used for yeast detection.…”

Section: Introductionmentioning

confidence: 99%

Modeling of an Impedimetric Biosensor with Ultrasonic-Assisted Cell Alignment for the Detection of Yeast

Tian

Xie

et al. 2022

Journal of Sensors

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Yeast is an integral part of our environment. The detection of yeast is of great significance in many fields. The impedance-based sensor with interdigital microelectrodes is a promising method to establish a simple and portable detection system. However, the positions of yeast cells greatly influence the impedance variation and eventually the sensitivity of detection. In this study, an impedimetric biosensor with ultrasonic-assisted cell alignment for yeast detection was proposed. In order to verify the feasibility of this biosensor, finite element modeling was conducted using the software COMSOL. The acoustic pressure field and the acoustic radiation force exerted on yeast cells were investigated. Cell positions in the microfluidic channel were determined using the fluid particle tracking module. After that, the impedance between the microelectrodes was calculated. Yeast suspensions with different cell concentrations were used as the tested samples for the simulation. The proposed sensor showed a higher sensitivity than the conventional impedimetric biosensor on which the cells were randomly located. It can be used for the detection of microorganisms. This finite element modeling provided an effective approach for the design of biosensors.

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Evaluation of Yeast Inoculated in Parallel to the Autochthonous Microbiota in Food Samples

Miguel¹,

Simões²,

Dias³

et al. 2021

Detection and Enumeration of Bacteria, Yeast, Viruses, and Protozoan in Foods and Freshwater

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Improved detection and enumeration of yeasts in wine by Cells-qPCR

Cited by 9 publications

References 27 publications

Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP

Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP

Modeling of an Impedimetric Biosensor with Ultrasonic-Assisted Cell Alignment for the Detection of Yeast

Evaluation of Yeast Inoculated in Parallel to the Autochthonous Microbiota in Food Samples

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