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Tytuł:
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16S ribosomal RNA-depletion PCR and its application in cause analysis of yogurt package shrinkage.
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Autorzy:
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Zhang H; State Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Centre of Dairy Biotechnology, Dairy Research Institute, Bright Dairy and Food Co. Ltd., Synergetic Innovation Centre of Food Safety and Nutrition, Shanghai 200436, China. Electronic address: .
You C; State Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Centre of Dairy Biotechnology, Dairy Research Institute, Bright Dairy and Food Co. Ltd., Synergetic Innovation Centre of Food Safety and Nutrition, Shanghai 200436, China.
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Źródło:
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Journal of dairy science [J Dairy Sci] 2022 Sep; Vol. 105 (9), pp. 7288-7297. Date of Electronic Publication: 2022 Aug 02.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Publication: Champaign, IL : American Dairy Science Association
Original Publication: Lancaster, Pa. [etc.]
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MeSH Terms:
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Food Microbiology*/methods
RNA, Ribosomal, 16S*/genetics
Yogurt*/microbiology
Animals ; DNA, Bacterial/analysis ; DNA, Ribosomal/genetics ; Food Packaging ; Polymerase Chain Reaction ; Streptococcus thermophilus/genetics
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Contributed Indexing:
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Keywords: 16S rRNA-depletion PCR; CRISPR-Cas9; Gluconobacter; shrinkage
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Substance Nomenclature:
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0 (DNA, Bacterial)
0 (DNA, Ribosomal)
0 (RNA, Ribosomal, 16S)
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Entry Date(s):
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Date Created: 20220805 Date Completed: 20220823 Latest Revision: 20220826
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Update Code:
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20240104
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DOI:
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10.3168/jds.2021-21575
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PMID:
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35931476
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Fermentative bacteria, the main microbiota in yogurt, interfere with the detection of unintended bacterial contaminants. The removal of fermentative bacteria and enrichment of unintended bacterial contaminants is a challenging task in bacterial detection. The present study developed a new 16S rRNA-depletion PCR for such enrichment and detection. Specifically, a single-guide RNA was designed and synthesized based on the 16S rRNA sequence of Streptococcus thermophilus, with the highest DNA abundance in the yogurt. The CRISPR-Cas9 system was used to specifically cleave and remove the genomic DNA of the fermentative bacteria, followed by PCR amplification. This method improved the detection sensitivity, simplified the operation steps, and reduced the detection cost of PCR analysis. We also used the 16S rRNA-depletion PCR to amplify and detect the unintended bacterial contaminants in yogurts with shrunken packages and analyzed the underlying reasons to prevent this issue of product quality.
(© 2022, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)