采用定量PCR法对微胶囊中添加全脂羊奶和菊粉型果聚糖的动物双歧杆菌乳亚种BB-12进行计数。

Bifidobacterium animalis ssp. lactis BB-12 enumeration by quantitative PCR assay in microcapsules with full-fat goat milk and inulin-type fructans.

作者信息

Verruck Silvani, Silva Kelly Justin, de Oliveira Santeli Helena, Scariot Mirella Christine, Venturelli Gustavo Luiz, Prudencio Elane Schwinden, Arisi Ana Carolina Maisonnave

机构信息

Dairy Technology Laboratory, Food Science and Technology Department, Agrarian Science Center, Federal University of Santa Catarina, Rod. Admar Gonzaga, 1346, Itacorubi, 88034-001 Florianópolis, SC, Brazil.

Molecular Biology Laboratory, Food Science and Technology Department, Agrarian Science Center, Federal University of Santa Catarina, Rod. Admar Gonzaga, 1346, Itacorubi, 88034-001 Florianópolis, SC, Brazil.

出版信息

Food Res Int. 2020 Jul;133:109131. doi: 10.1016/j.foodres.2020.109131. Epub 2020 Feb 27.

DOI:10.1016/j.foodres.2020.109131

PMID:32466908

Abstract

The current study was conducted to develop a quantitative polymerase chain reaction (qPCR) assay for Bifidobacterium animalis ssp. lactis BB-12 quantification in microcapsules matrix with full-fat goat milk and inulin-type fructans. DNA was isolated from milk, feed solutions (before spray drying) and microcapsules (after spray drying) using DNAzol. Two primer pairs targeting Bal-23S or Tuf sequences were evaluated by qPCR. The qPCR efficiency was higher (89.5%) using the Tuf primers than Bal-23S primers (84.8%). Tuf primer pair was able to selectively detect B. animalis ssp. lactis BB-12. After, quantification of bifidobacteria in the microcapsules matrix by Tuf qPCR assay was compared to conventional enumeration by plate counting. The analysis of probiotic feed solutions and microcapsules showed higher (P < 0.05) bacterial enumeration determined by Tuf qPCR assay compared to those obtained by plate counting. This qPCR assay was considered a rapid and sensitive alternative for the quantification of B. animalis ssp. lactis BB-12 in probiotic microcapsules compared to plate counting.

摘要

本研究旨在开发一种定量聚合酶链反应（qPCR）检测方法，用于对含有全脂羊奶和菊粉型果聚糖的微胶囊基质中的动物双歧杆菌亚种乳酸双歧杆菌BB - 12进行定量分析。使用DNAzol从牛奶、饲料溶液（喷雾干燥前）和微胶囊（喷雾干燥后）中提取DNA。通过qPCR评估了两对靶向Bal - 23S或Tuf序列的引物。使用Tuf引物时qPCR效率更高（89.5%），而使用Bal - 23S引物时效率为84.8%。Tuf引物对能够选择性地检测动物双歧杆菌亚种乳酸双歧杆菌BB - 12。之后，将通过Tuf qPCR检测法对微胶囊基质中的双歧杆菌进行定量分析的结果与通过平板计数进行的传统计数结果进行比较。对益生菌饲料溶液和微胶囊的分析表明，与平板计数法相比，通过Tuf qPCR检测法测定的细菌计数更高（P < 0.05）。与平板计数法相比，这种qPCR检测法被认为是一种快速且灵敏的方法，可用于对益生菌微胶囊中的动物双歧杆菌亚种乳酸双歧杆菌BB - 12进行定量分析。