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首页> 外文期刊>Artificial cells, nanomedicine, and biotechnology. >Use of qPCR for the analysis of population heterogeneity and dynamics during Lactobacillus delbrueckii spp. bulgaricus batch fculture
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Use of qPCR for the analysis of population heterogeneity and dynamics during Lactobacillus delbrueckii spp. bulgaricus batch fculture

机译：QPCR在乳酸杆菌SPP中的群体异质性和动态分析。保加利宫批量生活

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Direct molecular methods such as real-time polymerase chain reaction (qPCR) and propidium monoazide (PMA)-qPCR have been successfully used for quantifying viable microorganisms in the food industry. This study attempted to use qPCR and PMA-qPCR for quantifying Lactobacillus delbrueckii spp. bulgaricus sp1.1 physiological states. The qPCR standards of the 16S rRNA gene were employed to calibrate the qPCR assay, which contributed to an amplification efficiency of 98.42%. The number of copies of the 16S rRNA gene was linearly related to cell density, and this linear relationship was used to construct a quantitative curve ( R 2 =0.9981) with a detection limit of 15.1 colony-forming units mL ?1 ·reaction ?1 . qPCR in combination with an optimal PMA concentration (60?μM) helped in discriminating and quantifying the viable cells, without any interference by heat-killed cells. Compared with the conventional methods, the population heterogeneity of viable, culturable, dormant-like and membrane-permeabilized cells were well identified and quantified using qPCR during L . delbrueckii spp. bulgaricus sp1.1 batch culture. Despite the restriction in the enumeration of lysed cells, qPCR-based methods facilitated reliable identification and quantification of bacterial physiological states and provided additional knowledge on the dynamics of L . delbrueckii spp. bulgaricus sp1.1 physiological states.
机译：直接分子方法如实时聚合酶链式反应（QPCR）和单氮化丙锭（PMA）-QPCR已成功地用于量化食品工业中的可行微生物。该研究试图使用QPCR和PMA-QPCR来定量乳酸杆菌SPP。保加利宫SP1.1生理状态。 16S RRNA基因的QPCR标准物用于校准QPCR测定，这导致扩增效率为98.42％。 16S rRNA基因的拷贝数与细胞密度线性相关，并且这种线性关系用于构建定量曲线（R 2 = 0.9981），检测限为15.1个菌落形成单位mlα1·反应？1 。 QPCR与最佳PMA浓度（60Ωμm）组合有助于区分和定量活细胞，而不会通过热杀死的细胞干扰。与常规方法相比，使用QPCR在L期间鉴定并定量可行，培养，休眠样和膜渗透细胞的群体异质性。 delbrueckii spp。保加利宫SP1.1分批培养。尽管在裂解细胞的枚举中限制，但基于QPCR的方法促进了细菌生理状态的可靠鉴定和定量，并提供了关于L的动态的额外知识。 delbrueckii spp。保加利宫SP1.1生理状态。

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