Development of Certified Plasmid Reference Material for Hemolysin Gene vvhA of Vibrio vulnificus and It's Application in Aquatic Products Detection
Yuan Wei
Chen Leilei
Yang Jinyu
Zhou Qingxin
Qiu Jiying
Zhao Shuangzhi
Fu Enjun
Zhao Guoyan
Chen Xiangyan
Abstract：Due to lack of plasmid standard sample suitable for detection of Vibrio vulnificus in China,a ready-to-use certified plasmid reference material was developed herein.The recombinant plasmids of vvhA gene of V.vulnificus was constructed firstly and then their freeze-dried plasmid powders were prepared after sequen-cing verification and being detected by PCR and UV spectrophotometry.The uniformity test showed no signifi-cant difference between samples,and the uniformity was as expected.The stability test showed that the pre-pared plasmid samples could be stably preserved for 14 days at 4℃or 37℃,and for at least 12 months at-20℃.The overall results indicated that the uniformity and stability of the vvhA gene recombinant plasmid qualitative standard samples could meet the requirements of national qualitative standard samples,which pro-vided reliable reference material for rapid and high-throughput qualitative identification of V.vulnificus.The samples were used as positive control in the detection of 10 seafood samples such as fish and shellfish.and the results were confirmed to be accurate by traditional culture method.Above all,the qualitative standard samples of vvhA gene recombinant plasmid developed in this study had great potential in commercial application,laying an important foundation for its application in food detection.
Keywords：Vibrio vulnificusCertified plasmid reference materialAquatic productsDetection
Publication Date：2024-01-28
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：9( 147-155 )