南方科技大学知识苑(SUSTech KC): 一种基于CasRNA酶的一锅式核酸检测方法及应用

Title	一种基于CasRNA酶的一锅式核酸检测方法及应用
Alternative Title	One-pot nucleic acid detection method based on CasRNA enzyme and application
Author	蒋兴宇; 陈勇 ; 梅逸昕
First Inventor	蒋兴宇
Original applicant	南方科技大学
First applicant	南方科技大学
Address of First applicant	518055 广东省深圳市南山区西丽学苑大道1088号
Current applicant	南方科技大学
Address of Current applicant	518055 广东省深圳市南山区西丽学苑大道1088号 (广东,深圳,南山区)
First Current Applicant	南方科技大学
Address of First Current Applicant	518055 广东省深圳市南山区西丽学苑大道1088号 (广东,深圳,南山区)
Application Number	CN202011615558.2
Application Date	2020-12-30
Open (Notice) Number	CN112553307A
Date Available	2021-03-26
Status of Patent	实质审查
Legal Date	2021-04-13
Subtype	发明申请
SUSTech Authorship	First
Abstract	本发明提供一种基于Cas RNA酶的一锅式核酸检测方法及其应用。所述一锅式核酸检测方法包括：将待测溶液、RPA扩增单元、转录单元和含有Cas RNA酶的CRISPR单元混合，在扩增的同时实时监测反应体系的荧光强度，最终通过荧光动力学曲线实现对所述待测溶液中目标DNA的定量分析。所述一锅式核酸检测方法不但避免了扩增产物在转移过程中造成的气溶胶污染，防止出现假阳性结果，而且解决一步式“RPA扩增‑CRISPR‑Cas DNA酶”体系中，模板DNA不断被Cas DNA酶消化导致的检测时间长、检测灵敏度低等问题。
Other Abstract	The invention provides a one-pot nucleic acid detection method based on Cas RNA enzyme and application of the one-pot nucleic acid detection method. The one-pot nucleic acid detection method comprisesthe following steps of mixing a to-be-detected solution, an RPA amplification unit, a transcription unit and a CRISPR unit containing Cas RNA enzyme, monitoring the fluorescence intensity of a reaction system in real time while amplifying, and finally realizing quantitative analysis of target DNA in the to-be-detected solution through a fluorescence kinetic curve. According to the one-pot nucleicacid detection method, aerosol pollution caused by an amplification product in a transfer process is avoided, a false positive result is prevented, and the problems of long detection time, low detection sensitivity and the like caused by continuous digestion of template DNA by Cas DNA enzyme in a one-step RPA amplification=CRISPR-Cas DNA enzyme system are solved.
CPC Classification Number	C12Q1/6844 ; C12Q2521/507 ; C12Q2522/101 ; C12Q2531/119 ; C12Q2521/327 ; C12Q2521/107
IPC Classification Number	C12Q1/6844
INPADOC Legal Status	（ENTRY INTO FORCE OF REQUEST FOR SUBSTANTIVE EXAMINATION）[2021-04-13][CN]
INPADOC Patent Family Count	1
Extended Patent Family Count	1
Priority date	2020-12-30
Patent Agent	潘登
Agency	北京品源专利代理有限公司
URL	[Source Record]
Data Source	PatSnap
Document Type	Patent
Identifier	http://kc.sustech.edu.cn/handle/2SGJ60CL/534479
Department	Department of Biomedical Engineering
Recommended Citation GB/T 7714	蒋兴宇,陈勇,梅逸昕. 一种基于CasRNA酶的一锅式核酸检测方法及应用.

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