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 <i>tuf</i> gene used...
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| Format: | Article |
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MDPI AG
2021-12-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/10/12/3066 |
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| author | Xiankang Fan Xiefei Li Tao Zhang Jue Xu Zihang Shi Zhen Wu Jihuan Wu Daodong Pan Lihui Du |
| author_facet | Xiankang Fan Xiefei Li Tao Zhang Jue Xu Zihang Shi Zhen Wu Jihuan Wu Daodong Pan Lihui Du |
| author_sort | Xiankang Fan |
| collection | DOAJ |
| 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 <i>tuf</i> gene used in absolute quantification, the <i>tuf</i> 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<sup>3</sup>–1 × 10<sup>8</sup> 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. |
| first_indexed | 2024-03-10T04:08:22Z |
| format | Article |
| id | doaj.art-92ee2e36753446a9ba14fadc37006aab |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T04:08:22Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-92ee2e36753446a9ba14fadc37006aab2023-11-23T08:18:21ZengMDPI AGFoods2304-81582021-12-011012306610.3390/foods10123066A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented MilkXiankang Fan0Xiefei Li1Tao Zhang2Jue Xu3Zihang Shi4Zhen Wu5Jihuan Wu6Daodong Pan7Lihui Du8State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaNingbo Yifule Biotechnology Co., Ltd., Ningbo 315500, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo University, Ningbo 315211, ChinaThe 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 <i>tuf</i> gene used in absolute quantification, the <i>tuf</i> 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<sup>3</sup>–1 × 10<sup>8</sup> 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.https://www.mdpi.com/2304-8158/10/12/3066qPCRlactic acid bacteriayogurtlaser confocal microscope<i>tuf</i> geneflow cytometry |
| spellingShingle | Xiankang Fan Xiefei Li Tao Zhang Jue Xu Zihang Shi Zhen Wu Jihuan Wu Daodong Pan Lihui Du A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk Foods qPCR lactic acid bacteria yogurt laser confocal microscope <i>tuf</i> gene flow cytometry |
| 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 | qPCR lactic acid bacteria yogurt laser confocal microscope <i>tuf</i> gene flow cytometry |
| url | https://www.mdpi.com/2304-8158/10/12/3066 |
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