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1. CN101812528 - Switch mode aptamer probe and application thereof in tumor living cell and vital detection

Office China
Application Number 201010155148.4
Application Date 26.04.2010
Publication Number 101812528
Publication Date 25.08.2010
Grant Number 101812528
Grant Date 04.07.2012
Publication Kind B
IPC
C12Q 1/68
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
C12N 15/11
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
A61K 49/00
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
49Preparations for testing in vivo
G01N 21/64
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
CPC
A61K 48/0058
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
48Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
005characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
0058Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct
A61K 49/0054
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
49Preparations for testing in vivo
001Preparation for luminescence or biological staining
0013Luminescence
0017Fluorescence in vivo
005characterised by the carrier molecule carrying the fluorescent agent
0054Macromolecular compounds, i.e. oligomers, polymers, dendrimers
C12Q 1/6818
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6816characterised by the detection means
6818involving interaction of two or more labels, e.g. resonant energy transfer
C12Q 1/6886
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6876Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
6883for diseases caused by alterations of genetic material
6886for cancer
G01N 21/6428
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
G01N 21/6456
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
645Specially adapted constructive features of fluorimeters
6456Spatial resolved fluorescence measurements; Imaging
Applicants Hunan University
湖南大学
Inventors Guo Qiuping
王柯敏
He Xiaoxiao
石慧
Shi Hui
何晓晓
Wang Kemin
叶晓生
Wu Xu
伍旭
Ye Xiaosheng
郭秋平
Zhou Bing
周冰
Agents zhao hong
湖南兆弘专利事务所 43008
Title
(EN) Switch mode aptamer probe and application thereof in tumor living cell and vital detection
(ZH) 开关式核酸适体探针及其在肿瘤活细胞及活体检测中的应用
Abstract
(EN)
The invention belongs to the nucleic acid detection technical field, in particular discloses a switch mode aptamer probe and application thereof. The aptamer probe comprises a probe body as well as a fluorescence generating unit and a fluorescence extinguishing unit which are respectively connected at the two ends of the probe body, the probe body contains an aptamer segment with specificity identification function to target tumor cell and a nucleic acid segment forming complementary sequence with the aptamer segment, and the aptamer segment and the nucleic acid segment are connected to form a hairpin type structure by a connection segment with the length of 7-15nm; and the capacity of the nucleic acid segment competitive hybridization aptamer segment is weaken than that of the target tumor cell. The application of the probe of the invention can be at least one of specificity detection of tumor living cell in buffer solution, effective detection on tumor living cell in serum and real-time imaging and vital diction of tumor in moving organism. The probe of the invention has higher stability, specificity and sensibility, application operation thereof is simple, fast, sensible and specific, and cost is low.

(ZH)

本发明属于核酸检测技术领域,具体公开了一种开关式核酸适体探针及其应用,该核酸适体探针包括探针本体和分别连接于该探针本体两端的荧光发生单元和荧光熄灭单元,探针本体包含有对目标肿瘤细胞具有特异性识别功能的核酸适体片段和与该核酸适体片段形成互补序列的核酸片段,核酸适体片段和核酸片段通过一7~15nm长的连接片段连接形成一发夹型结构;核酸片段竞争性杂交核酸适体片段的能力弱于目标肿瘤细胞。本发明探针的应用可以是对缓冲液中肿瘤活细胞的特异性检测、对血清中肿瘤活细胞的有效检测及活动物体内肿瘤的实时荧光成像和活体检测中的至少一种。本发明的探针具有较高稳定性、特异性和灵敏性,其应用操作简单、快速、灵敏、特异、成本低。

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