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Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253

Probiotics are universally recognized for their health benefits, despite the fact that their effects depend on the strain. Identification and enumeration of probiotic strains are required prior to evaluating their effectiveness. Lacticaseibacillus rhamnosus X253 is a potential probiotic strain with...

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Autores principales: Zhao, Lei, Zhang, Dong, Liu, Yang, Zhang, Yi-Nan, Meng, Dong-Qing, Xu, Qiong, Zhong, Jiang, Jiang, Qiu-Yue, Zhao, Yu, Wang, Shi-Jie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9367767/
https://www.ncbi.nlm.nih.gov/pubmed/35954048
http://dx.doi.org/10.3390/foods11152282
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author Zhao, Lei
Zhang, Dong
Liu, Yang
Zhang, Yi-Nan
Meng, Dong-Qing
Xu, Qiong
Zhong, Jiang
Jiang, Qiu-Yue
Zhao, Yu
Wang, Shi-Jie
author_facet Zhao, Lei
Zhang, Dong
Liu, Yang
Zhang, Yi-Nan
Meng, Dong-Qing
Xu, Qiong
Zhong, Jiang
Jiang, Qiu-Yue
Zhao, Yu
Wang, Shi-Jie
author_sort Zhao, Lei
collection PubMed
description Probiotics are universally recognized for their health benefits, despite the fact that their effects depend on the strain. Identification and enumeration of probiotic strains are required prior to evaluating their effectiveness. Lacticaseibacillus rhamnosus X253 is a potential probiotic strain with antioxidant capacity. Comparative genomics and single nucleotide polymorphisms (SNPs) were used to identify a strain-specific locus within the holA gene for strain X253 that was distinct in 30 different L. rhamnosus strains. Using quantitative PCR, the primers and probe designed for the locus were able to distinguish L. rhamnosus X253 from the other 20 probiotic strains. The chosen locus remained stable over 19 generations. The sensitivity of the assay was 0.2 pg genomic DNA of L. rhamnosus X253, or 10(3) cfu/mL bacteria of this strain. In terms of repeatability and reproducibility, relative standard deviations (RSD) were less than 1% and 3%, respectively. Additionally, this assay achieved accurate enumerations of L. rhamnosus X253 in spiked milk and complex powder samples. The strain-specific assay could be used for quality control and compliance assessment of dairy products.
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spelling pubmed-93677672022-08-12 Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253 Zhao, Lei Zhang, Dong Liu, Yang Zhang, Yi-Nan Meng, Dong-Qing Xu, Qiong Zhong, Jiang Jiang, Qiu-Yue Zhao, Yu Wang, Shi-Jie Foods Article Probiotics are universally recognized for their health benefits, despite the fact that their effects depend on the strain. Identification and enumeration of probiotic strains are required prior to evaluating their effectiveness. Lacticaseibacillus rhamnosus X253 is a potential probiotic strain with antioxidant capacity. Comparative genomics and single nucleotide polymorphisms (SNPs) were used to identify a strain-specific locus within the holA gene for strain X253 that was distinct in 30 different L. rhamnosus strains. Using quantitative PCR, the primers and probe designed for the locus were able to distinguish L. rhamnosus X253 from the other 20 probiotic strains. The chosen locus remained stable over 19 generations. The sensitivity of the assay was 0.2 pg genomic DNA of L. rhamnosus X253, or 10(3) cfu/mL bacteria of this strain. In terms of repeatability and reproducibility, relative standard deviations (RSD) were less than 1% and 3%, respectively. Additionally, this assay achieved accurate enumerations of L. rhamnosus X253 in spiked milk and complex powder samples. The strain-specific assay could be used for quality control and compliance assessment of dairy products. MDPI 2022-07-30 /pmc/articles/PMC9367767/ /pubmed/35954048 http://dx.doi.org/10.3390/foods11152282 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhao, Lei
Zhang, Dong
Liu, Yang
Zhang, Yi-Nan
Meng, Dong-Qing
Xu, Qiong
Zhong, Jiang
Jiang, Qiu-Yue
Zhao, Yu
Wang, Shi-Jie
Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253
title Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253
title_full Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253
title_fullStr Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253
title_full_unstemmed Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253
title_short Quantitative PCR Assays for the Strain-Specific Identification and Enumeration of Probiotic Strain Lacticaseibacillus rhamnosus X253
title_sort quantitative pcr assays for the strain-specific identification and enumeration of probiotic strain lacticaseibacillus rhamnosus x253
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9367767/
https://www.ncbi.nlm.nih.gov/pubmed/35954048
http://dx.doi.org/10.3390/foods11152282
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