Preparation of plasmid DNA reference material for Listeria monocytogenes
ObjectiveTo develop a plasmid DNA reference material for rapid and comprehensive identification of Listeria monocytogenes.MethodsThe synthetic DNA fragment contained the hlyA, plcB and inlA, which were currently used for the detection of Listeria monocytogenes, was cloned into vector PUC57 to constr...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | zho |
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The Editorial Office of Chinese Journal of Food Hygiene
2021-05-01
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| Series: | Zhongguo shipin weisheng zazhi |
| Subjects: | |
| Online Access: | http://www.zgspws.com/zgspwszz/article/abstract/20210307?st=article_issue |
| _version_ | 1797976272342614016 |
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| author | PU Zenan LIN Xiaofeng YUAN Muyun Nusereti· Abudushalamu XU Longyan CHEN Yao |
| author_facet | PU Zenan LIN Xiaofeng YUAN Muyun Nusereti· Abudushalamu XU Longyan CHEN Yao |
| author_sort | PU Zenan |
| collection | DOAJ |
| description | ObjectiveTo develop a plasmid DNA reference material for rapid and comprehensive identification of Listeria monocytogenes.MethodsThe synthetic DNA fragment contained the hlyA, plcB and inlA, which were currently used for the detection of Listeria monocytogenes, was cloned into vector PUC57 to construct the plasmid reference material (pDNA Listeria). The quantity, homogeneity and stability of pDNA Listeria were evaluated by ultraviolet spectrophotometry (UV). real time quantitative polymerase chain reaction (qPCR) was used to evaluate the applicability of pDNA Listeria.ResultsThe results showed that the fixed value of pDNA Listeria was 29.85 μg/mL, which had traceable values, reliable homogeneity and good stability.The pDNA Listeria could be staored at -20 ℃ for more than one year. The pDNA Listeria also provided comparable sensitivity and reliability to the genomic references material.ConclusionThe pDNA Listeria could be used not only to identify Listeria monocytogenes but also to describe the biological characteristics such as virulence, pathogenicity, and invasiveness of Listeria monocytogenes at the same time. Importantly, the study proved the application of rapidly synthesized multiple targets plasmid serving as qPCR standard for pathogen identification. |
| first_indexed | 2024-04-11T04:48:22Z |
| format | Article |
| id | doaj.art-faf2d231eeee4d51ab19b41dc828bbf9 |
| institution | Directory Open Access Journal |
| issn | 1004-8456 |
| language | zho |
| last_indexed | 2024-04-11T04:48:22Z |
| publishDate | 2021-05-01 |
| publisher | The Editorial Office of Chinese Journal of Food Hygiene |
| record_format | Article |
| series | Zhongguo shipin weisheng zazhi |
| spelling | doaj.art-faf2d231eeee4d51ab19b41dc828bbf92022-12-27T09:23:07ZzhoThe Editorial Office of Chinese Journal of Food HygieneZhongguo shipin weisheng zazhi1004-84562021-05-01330327928410.13590/j.cjfh.2021.03.0071004-8456(2021)03-0279-06Preparation of plasmid DNA reference material for Listeria monocytogenesPU Zenan0LIN Xiaofeng1YUAN Muyun2Nusereti· Abudushalamu3XU Longyan4CHEN Yao5School of Laboratory and Biotechnology, Southern Medical University, Guangdong Guangzhou 510515, ChinaSchool of Laboratory and Biotechnology, Southern Medical University, Guangdong Guangzhou 510515, ChinaInspection and Quarantine Technology Center (IQTC), Guangdong Guangzhou 510623, ChinaSchool of Laboratory and Biotechnology, Southern Medical University, Guangdong Guangzhou 510515, ChinaInspection and Quarantine Technology Center (IQTC), Guangdong Guangzhou 510623, ChinaSchool of Laboratory and Biotechnology, Southern Medical University, Guangdong Guangzhou 510515, ChinaObjectiveTo develop a plasmid DNA reference material for rapid and comprehensive identification of Listeria monocytogenes.MethodsThe synthetic DNA fragment contained the hlyA, plcB and inlA, which were currently used for the detection of Listeria monocytogenes, was cloned into vector PUC57 to construct the plasmid reference material (pDNA Listeria). The quantity, homogeneity and stability of pDNA Listeria were evaluated by ultraviolet spectrophotometry (UV). real time quantitative polymerase chain reaction (qPCR) was used to evaluate the applicability of pDNA Listeria.ResultsThe results showed that the fixed value of pDNA Listeria was 29.85 μg/mL, which had traceable values, reliable homogeneity and good stability.The pDNA Listeria could be staored at -20 ℃ for more than one year. The pDNA Listeria also provided comparable sensitivity and reliability to the genomic references material.ConclusionThe pDNA Listeria could be used not only to identify Listeria monocytogenes but also to describe the biological characteristics such as virulence, pathogenicity, and invasiveness of Listeria monocytogenes at the same time. Importantly, the study proved the application of rapidly synthesized multiple targets plasmid serving as qPCR standard for pathogen identification.http://www.zgspws.com/zgspwszz/article/abstract/20210307?st=article_issuelisteria monocytogenessynthetic plasmid standardreal time fluorescent quantitative pcr |
| spellingShingle | PU Zenan LIN Xiaofeng YUAN Muyun Nusereti· Abudushalamu XU Longyan CHEN Yao Preparation of plasmid DNA reference material for Listeria monocytogenes Zhongguo shipin weisheng zazhi listeria monocytogenes synthetic plasmid standard real time fluorescent quantitative pcr |
| title | Preparation of plasmid DNA reference material for Listeria monocytogenes |
| title_full | Preparation of plasmid DNA reference material for Listeria monocytogenes |
| title_fullStr | Preparation of plasmid DNA reference material for Listeria monocytogenes |
| title_full_unstemmed | Preparation of plasmid DNA reference material for Listeria monocytogenes |
| title_short | Preparation of plasmid DNA reference material for Listeria monocytogenes |
| title_sort | preparation of plasmid dna reference material for listeria monocytogenes |
| topic | listeria monocytogenes synthetic plasmid standard real time fluorescent quantitative pcr |
| url | http://www.zgspws.com/zgspwszz/article/abstract/20210307?st=article_issue |
| work_keys_str_mv | AT puzenan preparationofplasmiddnareferencematerialforlisteriamonocytogenes AT linxiaofeng preparationofplasmiddnareferencematerialforlisteriamonocytogenes AT yuanmuyun preparationofplasmiddnareferencematerialforlisteriamonocytogenes AT nuseretiabudushalamu preparationofplasmiddnareferencematerialforlisteriamonocytogenes AT xulongyan preparationofplasmiddnareferencematerialforlisteriamonocytogenes AT chenyao preparationofplasmiddnareferencematerialforlisteriamonocytogenes |