Cargando…

A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk

The number of live lactic acid bacteria (LAB) is an important quality indicator for yogurt, the quantitative testing of LAB has become an important task in the evaluation of product quality and function. By analyzing and comparing the performance of 16S rRNA gene and tuf gene used in absolute quanti...

Descripción completa

Detalles Bibliográficos
Autores principales: Fan, Xiankang, Li, Xiefei, Zhang, Tao, Xu, Jue, Shi, Zihang, Wu, Zhen, Wu, Jihuan, Pan, Daodong, Du, Lihui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8700909/
https://www.ncbi.nlm.nih.gov/pubmed/34945617
http://dx.doi.org/10.3390/foods10123066
_version_ 1784620871686553600
author Fan, Xiankang
Li, Xiefei
Zhang, Tao
Xu, Jue
Shi, Zihang
Wu, Zhen
Wu, Jihuan
Pan, Daodong
Du, Lihui
author_facet Fan, Xiankang
Li, Xiefei
Zhang, Tao
Xu, Jue
Shi, Zihang
Wu, Zhen
Wu, Jihuan
Pan, Daodong
Du, Lihui
author_sort Fan, Xiankang
collection PubMed
description The number of live lactic acid bacteria (LAB) is an important quality indicator for yogurt, the quantitative testing of LAB has become an important task in the evaluation of product quality and function. By analyzing and comparing the performance of 16S rRNA gene and tuf gene used in absolute quantification, the tuf gene with copy number 1 was selected as the target gene of six LAB. By drawing a standard curve to achieve qualitative and quantitative detection of six strains of LAB, the detection range was found to be 1 × 10(3)–1 × 10(8) copies/µL. The traditional plate colony count and Flow Cytometry (FCM) were compared with the method of qPCR, which was used in this experiment. Meanwhile, the confocal laser microscope combined with STYO 9 and propidium iodide dyes was used to determine that the content of viable bacteria in the yogurt was more than 90%, which proved that the detection result using qPCR method was closer to the true level of LAB in yogurt. Compared with the existing methods, the method in this study allowed the qualitative and quantitative detection of the six kinds of LAB in yogurt, and the distribution of live and dead bacteria in yogurt could be calculated.
format Online
Article
Text
id pubmed-8700909
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-87009092021-12-24 A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk Fan, Xiankang Li, Xiefei Zhang, Tao Xu, Jue Shi, Zihang Wu, Zhen Wu, Jihuan Pan, Daodong Du, Lihui Foods Article The number of live lactic acid bacteria (LAB) is an important quality indicator for yogurt, the quantitative testing of LAB has become an important task in the evaluation of product quality and function. By analyzing and comparing the performance of 16S rRNA gene and tuf gene used in absolute quantification, the tuf gene with copy number 1 was selected as the target gene of six LAB. By drawing a standard curve to achieve qualitative and quantitative detection of six strains of LAB, the detection range was found to be 1 × 10(3)–1 × 10(8) copies/µL. The traditional plate colony count and Flow Cytometry (FCM) were compared with the method of qPCR, which was used in this experiment. Meanwhile, the confocal laser microscope combined with STYO 9 and propidium iodide dyes was used to determine that the content of viable bacteria in the yogurt was more than 90%, which proved that the detection result using qPCR method was closer to the true level of LAB in yogurt. Compared with the existing methods, the method in this study allowed the qualitative and quantitative detection of the six kinds of LAB in yogurt, and the distribution of live and dead bacteria in yogurt could be calculated. MDPI 2021-12-09 /pmc/articles/PMC8700909/ /pubmed/34945617 http://dx.doi.org/10.3390/foods10123066 Text en © 2021 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
Fan, Xiankang
Li, Xiefei
Zhang, Tao
Xu, Jue
Shi, Zihang
Wu, Zhen
Wu, Jihuan
Pan, Daodong
Du, Lihui
A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk
title A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk
title_full A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk
title_fullStr A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk
title_full_unstemmed A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk
title_short A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk
title_sort novel qpcr method for the detection of lactic acid bacteria in fermented milk
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8700909/
https://www.ncbi.nlm.nih.gov/pubmed/34945617
http://dx.doi.org/10.3390/foods10123066
work_keys_str_mv AT fanxiankang anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT lixiefei anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT zhangtao anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT xujue anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT shizihang anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT wuzhen anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT wujihuan anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT pandaodong anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT dulihui anovelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT fanxiankang novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT lixiefei novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT zhangtao novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT xujue novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT shizihang novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT wuzhen novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT wujihuan novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT pandaodong novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk
AT dulihui novelqpcrmethodforthedetectionoflacticacidbacteriainfermentedmilk