کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5740573 | 1616522 | 2017 | 10 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Cells-qPCR as a direct quantitative PCR method to avoid microbial DNA extractions in grape musts and wines Cells-qPCR as a direct quantitative PCR method to avoid microbial DNA extractions in grape musts and wines](/preview/png/5740573.png)
- Cells-qPCR is very sensitive and specific in grape musts and wine without dilution.
- It can be used with yeasts, lactic acid bacteria, and acetic acid bacteria.
- It does not require DNA extraction, overcoming inhibitors like polyphenols and ethanol.
- The presence of non-target microorganisms does not alter the method's specificity.
- This methodology is cheaper, simpler, and only takes 2Â h from samples to the results.
A novel quantitative PCR assay called Cells-qPCR has been developed for the rapid detection and quantification of yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) directly from grape must and wine that does not require DNA extraction. The assay was tested on Brettanomyces bruxellensis, Saccharomyces cerevisiae, Lactobacillus plantarum, Oenococcus oeni, Acetobacter aceti and Gluconobacter oxydans in culture media, and in white and red grape musts and wines. Standard curves were constructed from DNA and cells for the six target species in all the matrices. Good efficiencies were obtained for both when comparing DNA and cells standard curves. No reaction inhibition was observed between matrices for each species. Cells quantification was linear over a range of cell concentrations (7, 5 or 4 orders of magnitude) and detected as few as one cell per reaction in all the matrices. The developed Cells-qPCR assay is a robust, reliable, fast and specific method to detect and quantify different yeasts, LAB and AAB species in grape must and wine that avoids DNA extraction and overcomes the presence of inhibitors like polyphenols and ethanol.
Journal: International Journal of Food Microbiology - Volume 261, 16 November 2017, Pages 25-34