CN112236530A - 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒 - Google Patents

一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒 Download PDF

Info

Publication number
CN112236530A
CN112236530A CN201980037873.0A CN201980037873A CN112236530A CN 112236530 A CN112236530 A CN 112236530A CN 201980037873 A CN201980037873 A CN 201980037873A CN 112236530 A CN112236530 A CN 112236530A
Authority
CN
China
Prior art keywords
nucleic acid
cas12b
detection
acid molecule
protein
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201980037873.0A
Other languages
English (en)
Inventor
李诗渊
王金
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tolo Biotech Co ltd
Original Assignee
Tolo Biotech Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tolo Biotech Co ltd filed Critical Tolo Biotech Co ltd
Publication of CN112236530A publication Critical patent/CN112236530A/zh
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • C12Q1/683—Hybridisation assays for detection of mutation or polymorphism involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases RNAses, DNAses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPRs]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/10—Nucleotidyl transfering
    • C12Q2521/107—RNA dependent DNA polymerase,(i.e. reverse transcriptase)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q2527/00—Reactions demanding special reaction conditions
    • C12Q2527/125—Specific component of sample, medium or buffer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q2545/00—Reactions characterised by their quantitative nature
    • C12Q2545/10—Reactions characterised by their quantitative nature the purpose being quantitative analysis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

本发明提供了一种耐高温Cas蛋白的用途及靶标核酸分子的检测体系和试剂盒，具体地，本发明提供一种用于检测靶标核酸分子的反应体系，所述反应体系包括：向导RNA、Cas12b(旧称C2c1)、核酸探针，反应完成后对其进行检测。另外，通过与核酸扩增技术(如LAMP等)的结合，该检测方法的灵敏度可以得到大幅度提高。本发明提供的检测体系可用于快速检测病原微生物、基因突变、单核苷酸多态性和特异靶标DNA等，并且可以对核酸样品进行定量。

Description

PCT国内申请，说明书已公开。

Claims (15)

  1. PCT国内申请，权利要求书已公开。
CN201980037873.0A 2018-06-03 2019-06-03 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒 Pending CN112236530A (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CN201810560284.8A CN110551800B (zh) 2018-06-03 2018-06-03 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒
CN2018105602848 2018-06-03
PCT/CN2019/089856 WO2019233385A1 (zh) 2018-06-03 2019-06-03 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Publications (1)

Publication Number Publication Date
CN112236530A true CN112236530A (zh) 2021-01-15

Family

ID=68735376

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201810560284.8A Active CN110551800B (zh) 2018-06-03 2018-06-03 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒
CN201980037873.0A Pending CN112236530A (zh) 2018-06-03 2019-06-03 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN201810560284.8A Active CN110551800B (zh) 2018-06-03 2018-06-03 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Country Status (10)

Country Link
US (1) US20210238666A1 (zh)
EP (1) EP3805403A4 (zh)
JP (1) JP7431453B2 (zh)
KR (2) KR20230116944A (zh)
CN (2) CN110551800B (zh)
AU (1) AU2019280305B2 (zh)
BR (1) BR112020024749A2 (zh)
CA (1) CA3102438C (zh)
WO (1) WO2019233385A1 (zh)
ZA (1) ZA202008068B (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110551800A (zh) * 2018-06-03 2019-12-10 上海吐露港生物科技有限公司 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2022501039A (ja) * 2018-09-20 2022-01-06 インスティテュート オブ ズーオロジー、チャイニーズ アカデミー オブ サイエンシーズ 核酸を検出する方法
CN111187804A (zh) * 2020-02-20 2020-05-22 国家卫生健康委科学技术研究所 一种基于CRISPR/Cas12a的肺炎支原体核酸快速检测试剂盒及其检测方法
CN111455046A (zh) * 2020-03-13 2020-07-28 深圳市罗湖区人民医院 一种adgrg6增强子突变的核酸检测方法及其试剂盒
CN111505275B (zh) * 2020-03-20 2023-03-31 浙江工业大学 一种基于Cas9核酸等温扩增的免疫层析多重基因检测方法
CN111733216B (zh) * 2020-06-22 2023-03-28 山东舜丰生物科技有限公司 一种提高目标核酸检测效率的方法
CN111778318B (zh) * 2020-07-10 2023-01-10 清华大学深圳国际研究生院 一种基于CRISPR/Cas系统检测核酸分子的方法及系统
CN111926117B (zh) * 2020-08-18 2022-09-13 交弘生物科技(上海)有限公司 SARS-CoV-2病毒核酸等温快速检测试剂盒及检测方法
CN116024314A (zh) * 2020-09-18 2023-04-28 山东舜丰生物科技有限公司 基于crispr技术进行靶核酸多重检测的方法
CN114507716A (zh) * 2020-11-16 2022-05-17 北京迅识科技有限公司 检测样品中靶核酸的方法
WO2022148450A1 (en) * 2021-01-08 2022-07-14 Wuhan University Compositions and methods for instant nucleic acid detection
CN113957164B (zh) * 2021-10-29 2023-05-23 上海市质量监督检验技术研究院 一种婴儿配方乳粉中克罗诺杆菌属的CRISPR One Pot检测方法及其试剂盒
CN114634972B (zh) * 2022-05-19 2022-08-26 舜丰生物科技(海南)有限公司 利用Cas酶进行核酸检测的方法
CN115141826B (zh) * 2022-06-01 2024-01-05 安徽省农业科学院畜牧兽医研究所 Rpa引物对及其应用、可视化检测pcv4的试剂盒及其应用和检测pcv4方法
CN115404268B (zh) * 2022-10-11 2023-04-07 芜湖森爱驰生物科技有限公司 一种sry基因检测探针和试剂盒
CN115975985B (zh) * 2022-10-14 2023-09-26 湖北大学 一种中高温Argonaute蛋白在特异性切割靶核酸中的应用
CN115820932B (zh) * 2022-10-18 2023-12-22 华中科技大学协和深圳医院 一种基于crispr系统反式切割和石墨烯场效应晶体管的核酸检测方法
CN116574841B (zh) * 2023-06-27 2023-12-15 广西大学 一种非特异CrRNA的CRISPR/Cas12a切割体系的多重病原菌的检测方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107488710A (zh) * 2017-07-14 2017-12-19 上海吐露港生物科技有限公司 一种Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015116686A1 (en) * 2014-01-29 2015-08-06 Agilent Technologies, Inc. Cas9-based isothermal method of detection of specific dna sequence
EP3294877A1 (en) * 2015-05-15 2018-03-21 Pioneer Hi-Bred International, Inc. Rapid characterization of cas endonuclease systems, pam sequences and guide rna elements
US10337051B2 (en) * 2016-06-16 2019-07-02 The Regents Of The University Of California Methods and compositions for detecting a target RNA
CN117487776A (zh) * 2017-11-01 2024-02-02 加利福尼亚大学董事会 Casz组合物和使用方法
US10253365B1 (en) * 2017-11-22 2019-04-09 The Regents Of The University Of California Type V CRISPR/Cas effector proteins for cleaving ssDNAs and detecting target DNAs
CN110551800B (zh) * 2018-06-03 2022-08-19 上海吐露港生物科技有限公司 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107488710A (zh) * 2017-07-14 2017-12-19 上海吐露港生物科技有限公司 一种Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒
CN111094588A (zh) * 2017-07-14 2020-05-01 上海吐露港生物科技有限公司 一种Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110551800A (zh) * 2018-06-03 2019-12-10 上海吐露港生物科技有限公司 一种耐高温Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒

Also Published As

Publication number Publication date
CA3102438C (en) 2023-01-24
KR20230116944A (ko) 2023-08-04
WO2019233385A1 (zh) 2019-12-12
EP3805403A4 (en) 2022-05-04
CN110551800B (zh) 2022-08-19
BR112020024749A2 (pt) 2021-03-23
JP7431453B2 (ja) 2024-02-15
US20210238666A1 (en) 2021-08-05
CA3102438A1 (en) 2019-12-12
JP2021528098A (ja) 2021-10-21
KR20210039989A (ko) 2021-04-12
ZA202008068B (en) 2022-03-30
CN110551800A (zh) 2019-12-10
EP3805403A1 (en) 2021-04-14
AU2019280305B2 (en) 2023-07-20
AU2019280305A1 (en) 2021-01-21

Similar Documents

Publication Publication Date Title
JP7431453B2 (ja) 耐高温Ｃａｓタンパク質の使用および標的核酸分子の検出方法とキット
CN111094588B (zh) 一种Cas蛋白的用途及靶标核酸分子的检测方法和试剂盒
CN107849603B (zh) 利用有限核苷酸组成的引物扩增
EP2694682B1 (en) High through-put analysis of transgene borders
EP3102702A1 (en) Error-free sequencing of dna
US20130059734A1 (en) Epigenetic analysis
MX2011009736A (es) Deteccion molecular basada en tecnologia abscript.
EP2432899A1 (en) Sorting asymmetrically tagged nucleic acids by selective primer extension
US20090215034A1 (en) Method for selectively detecting subsets of nucleic acid molecules
US10023908B2 (en) Nucleic acid amplification method using allele-specific reactive primer
US20230031001A1 (en) Method for detecting target nucleic acid, method for detecting nucleic acid-binding molecule, and method for evaluating nucleic acid-binding ability
US20090131275A1 (en) Method For Preparing Genome Library, And Genome Library Prepared By The Method
WO2011101744A2 (en) Region of interest extraction and normalization methods
WO1996014406A1 (en) Method of preparing oligonucleotide probes or primers, vector therefor and use thereof
RU2800778C2 (ru) Применение высокотермостойкого cas-белка и способ и набор реактивов для обнаружения целевой молекулы нуклеиновой кислоты
WO2013078318A1 (en) High through-put analysis of transgene borders
JP2006271250A (ja) 塩基配列決定方法
Thomas Polymerase chain reaction and its various modifications In: Winter School on Vistas in Marine Biotechnology 5th to 26th October 2010

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination