Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus
ObjectiveA dual real-time polymerase chain reaction (PCR) method for simultaneous detection of two virulence genes tdh and trh of Vibrio parahaemolyticus was established, and the virulence genes carried by 2 771 strains of foodborne Vibrio parahaemolyticus in China were comprehensively detected.Meth...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | zho |
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The Editorial Office of Chinese Journal of Food Hygiene
2022-01-01
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| Series: | Zhongguo shipin weisheng zazhi |
| Subjects: | |
| Online Access: | http://www.zgspws.com/zgspwszz/article/abstract/20220111?st=article_issue |
| _version_ | 1797976219696758784 |
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| author | BAI Yao LI Bin LI Fengqin YANG Dajin XU Jin DONG Yinping WANG Wei YAN Lin JIANG Tao |
| author_facet | BAI Yao LI Bin LI Fengqin YANG Dajin XU Jin DONG Yinping WANG Wei YAN Lin JIANG Tao |
| author_sort | BAI Yao |
| collection | DOAJ |
| description | ObjectiveA dual real-time polymerase chain reaction (PCR) method for simultaneous detection of two virulence genes tdh and trh of Vibrio parahaemolyticus was established, and the virulence genes carried by 2 771 strains of foodborne Vibrio parahaemolyticus in China were comprehensively detected.MethodsAccording to the tdh and trh virulence genes of Vibrio parahaemolyticus, PCR primers and fluorescent probes were designed respectively, the real-time PCR reaction system and reaction procedure were optimized, and a dual real-time PCR detection method which can detect the two virulence genes at the same time was established. The virulence genes carried by 2 771 strains of foodborne Vibrio parahaemolyticus isolated in 2015 and 2016 were detected by the established method, and compared with the result of PCR method to evaluate the sensitivity, accuracy and specificity of the method.ResultsThe established dual fluorescence PCR method could detect both tdh and trh virulence genes at the same time, and its sensitivity was 1.5×10-4 ng/μL. The accuracy and specificity were 100%. In 2015, the carrying rates of tdh and trh genes in foodborne Vibrio parahaemolyticus in China were 0.26% (3/1 137) and 1.67%(19/1 137) respectively, and were 0.24%(4/1 634) and 0.43%(7/1 634) in 2016 respectively.ConclusionIn this study, a dual real-time PCR method for simultaneous detection of tdh and trh virulence genes of Vibrio parahaemolyticus was established, which could quickly and accurately screen the virulence genes of Vibrio parahaemolyticus. The carrying rate of tdh and trh virulence genes of Vibrio parahaemolyticus isolated from food in China was low. The dual real-time PCR method can be applied to the study of the pathogenicity of Vibrio parahaemolyticus in food, and provide scientific data for the risk assessment of dietary exposure to Vibrio parahaemolyticus in China. |
| first_indexed | 2024-04-11T04:47:37Z |
| format | Article |
| id | doaj.art-42e713d0cb6f4a78a680523dfec9e0ce |
| institution | Directory Open Access Journal |
| issn | 1004-8456 |
| language | zho |
| last_indexed | 2024-04-11T04:47:37Z |
| publishDate | 2022-01-01 |
| publisher | The Editorial Office of Chinese Journal of Food Hygiene |
| record_format | Article |
| series | Zhongguo shipin weisheng zazhi |
| spelling | doaj.art-42e713d0cb6f4a78a680523dfec9e0ce2022-12-27T09:24:30ZzhoThe Editorial Office of Chinese Journal of Food HygieneZhongguo shipin weisheng zazhi1004-84562022-01-013401545910.13590/j.cjfh.2022.01.0111004-8456(2022)01-0054-06Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticusBAI Yao0LI Bin1LI Fengqin2YANG Dajin3XU Jin4DONG Yinping5WANG Wei6YAN Lin7JIANG Tao8Key Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaKey Laboractory of Food Safety Assessment of Ministry of Health, China National Center for Food Safety Assessment, Beijing 100021, ChinaObjectiveA dual real-time polymerase chain reaction (PCR) method for simultaneous detection of two virulence genes tdh and trh of Vibrio parahaemolyticus was established, and the virulence genes carried by 2 771 strains of foodborne Vibrio parahaemolyticus in China were comprehensively detected.MethodsAccording to the tdh and trh virulence genes of Vibrio parahaemolyticus, PCR primers and fluorescent probes were designed respectively, the real-time PCR reaction system and reaction procedure were optimized, and a dual real-time PCR detection method which can detect the two virulence genes at the same time was established. The virulence genes carried by 2 771 strains of foodborne Vibrio parahaemolyticus isolated in 2015 and 2016 were detected by the established method, and compared with the result of PCR method to evaluate the sensitivity, accuracy and specificity of the method.ResultsThe established dual fluorescence PCR method could detect both tdh and trh virulence genes at the same time, and its sensitivity was 1.5×10-4 ng/μL. The accuracy and specificity were 100%. In 2015, the carrying rates of tdh and trh genes in foodborne Vibrio parahaemolyticus in China were 0.26% (3/1 137) and 1.67%(19/1 137) respectively, and were 0.24%(4/1 634) and 0.43%(7/1 634) in 2016 respectively.ConclusionIn this study, a dual real-time PCR method for simultaneous detection of tdh and trh virulence genes of Vibrio parahaemolyticus was established, which could quickly and accurately screen the virulence genes of Vibrio parahaemolyticus. The carrying rate of tdh and trh virulence genes of Vibrio parahaemolyticus isolated from food in China was low. The dual real-time PCR method can be applied to the study of the pathogenicity of Vibrio parahaemolyticus in food, and provide scientific data for the risk assessment of dietary exposure to Vibrio parahaemolyticus in China.http://www.zgspws.com/zgspwszz/article/abstract/20220111?st=article_issuereal-time polymerase chain reactionvibrio parahaemolyticusvirulence genesdetection |
| spellingShingle | BAI Yao LI Bin LI Fengqin YANG Dajin XU Jin DONG Yinping WANG Wei YAN Lin JIANG Tao Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus Zhongguo shipin weisheng zazhi real-time polymerase chain reaction vibrio parahaemolyticus virulence genes detection |
| title | Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus |
| title_full | Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus |
| title_fullStr | Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus |
| title_full_unstemmed | Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus |
| title_short | Establishment and application of dual real-time PCR for detection of virulence genes of Vibrio parahaemolyticus |
| title_sort | establishment and application of dual real time pcr for detection of virulence genes of vibrio parahaemolyticus |
| topic | real-time polymerase chain reaction vibrio parahaemolyticus virulence genes detection |
| url | http://www.zgspws.com/zgspwszz/article/abstract/20220111?st=article_issue |
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