摘要
为了有效预防宫颈癌的发生以及更好的术后跟踪治疗,实现对HPV病毒DNA快速可靠的检测是至关重要的。与传统方法相比,通过缩短检测时间和减少劳动力来进一步简化HPV病毒DNA检测仍然是一个挑战。该研究提出了一种基于CRISPR/Cas12a对基因的SERS检测方法。主要利用目标病毒核酸与Cas12a、crRNA形成三元复合体,开启切割体系中的底物单链DNA(ssDNA)。ssDNA可以连接探针1(AuNPs@MBN@DNA1)和探针2(AuNPs@MBN@DNA2)。探针1和探针2是否被桥连可引起SERS信号的变化,即可知待检样品中是否含有目标核酸。利用特异性的crRNA可以实现对病毒核酸的准确快速检测。该传感方法可在40 min内实现对HPV基因的高灵敏度检测。由于DNA探针和crRNA设计的灵活性,该CRISPR/Cas-SERS传感系统可以扩展成一种通用的基因检测工具,有望在体外诊断领域和检测中具有广阔的应用前景。
In order to effectively prevent the occurrence of cervical cancer and better postoperative follow-up treatment,it is crucial to achieving rapid and reliable detection of HPV virus DNA.The simplification of HPV viral DNA testing by reducing testing time and labor compared to traditional methods remains a challenge.Here,we propose a SERS gene detection method based on CRISPR/Cas12a.It mainly uses the target viral nucleic acid to form a ternary complex with Cas12aand crRNA,and cuts the substrate single-stranded DNA(ssDNA)in the cutting system.The ssDNA can ligate probe 1(AuNPs@MBN@DNA1)and probe 2(AuNPs@MBN@DNA2).Whether probe 1and probe 2are bridged can cause the change in the SERS signal,and it can be known whether the sample to be tested contains the target nucleic acid.The use of specific crRNA can achieve accurate and rapid detection of viral nucleic acid.The sensing method can achieve high-sensitivity detection of HPV genes within 40minutes.Due to the flexibility of DNA probe and crRNA design,this CRISPR/Cas-SERS sensing system can be extended into a general gene detection tool,which is expected to have broad application prospects in the field of in vitro diagnostics and detection.
作者
苏艾玲
曹修冕
徐蔚青
徐抒平
SU Ai-ling;CAO Xiu-mian;XU Wei-qing;XU Shu-ping(Key Laboratory for Supramolecular Structure and Materials of Ministry of Education,College of Chemistry,Jilin University,Changchun 130012,China;Center for Supramolecular Chemical Biology,College of Chemistry,Jilin University,Changchun 130012,China;College of Physics,Jilin University,Changchun 130012,China)
出处
《光谱学与光谱分析》
SCIE
EI
CAS
CSCD
北大核心
2023年第S01期191-192,共2页
Spectroscopy and Spectral Analysis
基金
国家自然科学基金项目(21873039)
应用光学国家重点实验室开放项目(SKLAO2021001A14)
吉林大学交叉融合创新项目(JLUXKJC2020106)
吉林省科技发展计划项目(20220101046JC)资助
