基于氟化物核糖开关转录反应体系和CRISPR/Cas12a检测南极磷虾中氟含量

Detection of fluoride content in Euphausia superba based on fluoride riboswitch transcription system and CRISPR/Cas12a

中文关键词:  氟化物核糖开关  CRISPR/Cas12a  氟含量检测  南极磷虾

英文关键词:fluoride riboswitch  clustered regularly interspaced short palindromic repeats-associated protein 12a  fluoride content detection  Euphausia superba

中文摘要:

      目的 建立氟化物核糖开关转录反应体系以及成簇的规律间隔的短回文重复序列/Cas12a蛋白(clustered regularly interspaced short palindromic repeats-associated protein 12a, CRISPR/Cas12a)检测氟含量的方法。方法 本研究利用氟化物核糖开关在有氟或无氟下转录全长或非全长RNA并与CRISPR/Cas12a系统相结合, 通过荧光法读取实验结果。对优化的CRISPR/Cas12a检测体系进行选择性和干扰性分析实验, 并用于南极磷虾氟含量的检测。结果 本研究成功建立了基于氟化物核糖开关转录反应体系CRISPR/Cas12a荧光检测方法, 可在1.5 h内完成对样品的检测, 检测结果与国家标准方法氟离子选择电极法基本保持一致。结论 本研究建立的基于氟化物核糖开关转录反应和CRISPR/Cas12a的南极磷虾中氟含量检测方法具有高特异性和准确性, 在氟含量的检测方面具有应用价值。

英文摘要:

      Objective To establish a method to detect fluoride content based on the fluoride riboswitch transcription system and clustered regularly interspaced short palindromic repeats-associated protein 12a (CRISPR/Cas12a). Methods The system utilized fluoride riboswitch to regulate transcription of full-length or truncated RNA in the presence/absence of fluoride, which was then detected by CRISPR/Cas12a-mediated fluorescence signal. Selectivity and interference analyses were conducted to optimize CRISPR/Cas12a detection system, and the method was applied to measure fluoride content in Euphausia superba. Results This study successfully established a fluorescence detection method based on a fluoride riboswitch transcription reaction system and CRISPR/Cas12a, which completed sample detection within 1.5 h with the results consistent with those obtained by the national standard fluoride ion-selective electrode method. Conclusion The fluoride detection method established in this study for Euphausia superba based on a fluoride riboswitch transcription reaction system and CRISPR/Cas12a, demonstrates high specificity and accuracy, showing potential for fluoride content analysis.