摘要
目的探讨适用于创伤弧菌的特异性引物,通过微流控芯片技术进行一站式检测,为快速检测出病原体奠定基础。方法通过NCBI网站获取靶基因序列,采用MEGA7.0软件对齐后设计出19对引物,BLAST确定引物的特异度,再通过引物性能、灵敏度、快速变温实验筛选适用于微流控的引物,最后对引物的特异度与灵敏度进行评价。结果成功筛选出1对适用于微流控的引物vvhA10,微流控检测结果发现其特异度与灵敏度较高。结论筛选出适用于微流控芯片的引物,用于全自动微流控检测可以满足应急检测或现场快速检测等方面的使用需求。
Objective To explore the specific primers suitable for Vibrio vulnificus and to perform one-stop detection by microfluidic chip technology,so as to lay the foundation for rapid detection of pathogens.Methods The target gene sequence was obtained from the NCBI website,and 19 pairs of primers were designed after alignment by MEGA 7.0 software.The specificity of the primers was determined by BLAST,and the primers suitable for microfluidic were screened by primer performance,sensitivity and rapid temperature change test.Results A pair of primers vvhA10 was successfully screened.The results of microfluidic detection showed that its specificity and sensitivity were high.Conclusion The selected primers are suitable for automatic microfluidic chip detection,which can meet the needs of emergency detection or rapid detection in the field.
作者
张小红
禹乐
朱启淦
孟加榕
ZHANG Xiaohong;YU Le;ZHU Qigan;MENG Jiarong(Department of Pathology,909th Hospital/Southeast Hospital Affiliated to Xiamen University,Zhangzhou,Fujian 363000,China)
出处
《检验医学与临床》
CAS
2024年第11期1579-1585,1594,共8页
Laboratory Medicine and Clinic
基金
福建省漳州市自然科学基金项目(ZZ2021J09)
福建省自然科学基金项目(2023J011843,2023J011841)
联勤保障部队第九〇九医院自主项目(22MS001)。
关键词
微流控芯片
创伤弧菌
引物筛选
荧光定量聚合酶链反应
即时检测
microfluidic chip
Vibrio vulnificus
screening of primers
fluorescence quantitative polymerase chain reaction
point-of-care testing
