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高分辨率熔解分析在富含益生菌的食品基质中变种鉴定的潜力

High-Resolution Melting Analysis Potential for var. Authentication in Probiotic-Enriched Food Matrices.

作者信息

Borkowska Monika, Kułakowski Michał, Myszka Kamila

机构信息

Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-637 Poznan, Poland.

出版信息

BioTech (Basel). 2024 Nov 14;13(4):48. doi: 10.3390/biotech13040048.

Abstract

To date, the only probiotic yeast with evidence of health-promoting effects is var. . The expanded market including dietary supplements and functional foods supplemented with var. creates an environment conductive to food adulterations, necessitating rapid testing to verify product probiotic status. Herein, qPCR-HRM analysis was tested for probiotic yeast identification. The effectiveness of the primer pairs' set was examined, designed to amplify heterogeneous regions in (a) rDNA sequences previously designed to identify food-derived yeast and (b) genes associated with physiological and genotypic divergence of var. Preliminary tests of amplicons' differentiation power enabled the selection of interspecies sequences for and ITS and genus-specific sequences , , and The multi-fragment qPCR-HRM analysis was sufficient for culture-dependent var. identification and proved effective in the authentication of dietary supplements' probiotic composition. The identification of var. in complex microbial mixtures of kefir succeeded with more specific intragenus sequences and The predominance of var. in the tested matrices, quantitatively corresponded to the probiotic-enriched food, was crucial for identification with qPCR-HRM analysis. Considering the reported assumptions, qPCR-HRM analysis is an appropriate tool for verifying probiotic-enriched food.

摘要

迄今为止,唯一有促进健康作用证据的益生菌酵母是 变种。包括补充了 变种的膳食补充剂和功能性食品在内的扩大市场,营造了一个易于食品掺假的环境,因此需要进行快速检测以核实产品的益生菌状态。在此,对qPCR-HRM分析进行了测试,用于鉴定益生菌酵母。对引物对组的有效性进行了检验,这些引物对旨在扩增(a)先前设计用于鉴定食品来源酵母的rDNA序列中的异质区域,以及(b)与 变种的生理和基因型差异相关的基因。对扩增子区分能力的初步测试使得能够选择 和ITS的种间序列以及属特异性序列 、 、 和 。多片段qPCR-HRM分析足以用于依赖培养的 变种鉴定,并被证明在验证膳食补充剂的益生菌成分方面有效。利用更具特异性的属内序列 和 ,成功地在开菲尔的复杂微生物混合物中鉴定出了 变种。在测试基质中 变种的优势,在数量上与富含益生菌的食物相对应,这对于通过qPCR-HRM分析进行鉴定至关重要。考虑到所报道的假设,qPCR-HRM分析是验证富含益生菌食物的合适工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5bae/11586983/a99dde1f5d84/biotech-13-00048-g002.jpg

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