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一种基于CRISPR-Cas13a的特异性核酸片段的检测方法及探针

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摘要

本发明公开了一种基于CRISPR‑Cas13a的特异性核酸片段的检测方法，该方法的检测体系中含有：crRNA、Cas13a、探针；该方法的特征在于：所述的探针包含1段RNA序列，该RNA序列的两端分别连接1段DNA序列；所述探针上的核苷酸均进行硫代磷酸骨架的修饰，以在相邻核苷酸之间形成硫代磷酸酯键；所述RNA序列上的若干个核苷酸还进行了二甲氧基修饰，并且，所述的RNA序列上至少含有两个连续的未进行二甲氧基修饰的核苷酸。所述的DNA序列含有6个脱氧核糖核苷酸。所述的RNA序列含有为8‑12个核糖核苷酸。本发明设计的探针稳定高，荧光背景值低。

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公开/公告号CN110066865A

专利类型发明专利
公开/公告日2019-07-30

原文格式PDF
申请/专利权人常州桐树生物科技有限公司;
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申请/专利号CN201910359662.0
发明设计人蔡微菁;孙子豪;
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申请日2019-04-30
分类号
代理机构上海元好知识产权代理有限公司;
代理人贾慧琴
地址 213149 江苏省常州市常州西太湖科技产业园长扬路9号B2栋2楼
入库时间 2024-02-19 11:23:21

法律信息

法律状态公告日

法律状态信息

法律状态
2019-08-23

实质审查的生效 IPC(主分类):C12Q1/6876 申请日:20190430

实质审查的生效
2019-07-30

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公开

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1. 一种基于CRISPR-Cas13a的特异性核酸片段的检测方法及探针 [P] . 中国专利： CN110066865A . 2019-07-30
2. 一种基于短链核酸探针和双链特异性核酸内切酶的高特异性microRNA荧光检测方法 [P] . 中国专利： CN107267604A . 2017-10-20
3. pencicillin-binding protein, nucleic acid, antibody or antibody fragment, medicament, pharmaceutical composition, uses of at least one penicillin-binding protein or fragment or variant or fragment thereof, of at least one nucleic acid and of at least one antibody or antibody fragment, in vitro antibody detection methods directed against neisseria meningitidis infection from a penicillin-binding protein, nucleic acid, antibody or an antibody fragment, pharmaceutical composition of at least one penicillin-binding protein or fragment or variant or variant thereof, of at least one nucleic acid and at least one antibody directed against neisseria meningitidis infection from a biological sample of a mammal and in vitro diagnosis of neisseria miningitidis infection from a biological sample from a mammal and monoclonal antibody [P] . BRPI0610802A2 . 2016-11-16

机译：青霉素结合蛋白，核酸，抗体或抗体片段，药物，药物组合物，至少一种青霉素结合蛋白或其片段或变异体或片段，至少一种核酸和至少一种抗体或抗体片段的用途，针对脑膜炎奈瑟菌的青霉素结合蛋白，核酸，抗体或抗体片段感染的体外抗体检测方法，至少一种青霉素结合蛋白或其片段或变异体或变异体，至少一种核酸的药物组合物酸和至少一种针对哺乳动物生物样品中脑膜炎奈瑟氏菌感染的抗体以及来自哺乳动物生物样品中的矿业性奈瑟氏球菌感染的体外诊断和单克隆抗体
4. isolated antigen binding region that is specific for cd38, isolated antibody or a functional fragment thereof that is specific for cd38, isolated antibody or a functional fragment thereof, isolated immunoglobulin, heavy chain variable of an isolated antigen binding region, variable light chain from an isolated antigen binding region, isolated nucleic acid sequence, nucleic acid sequence encoding a variable heavy chain from an isolated antigen binding region, nucleic acid sequence, vector, isolated cell, pharmaceutical composition, use of the pharmaceutical composition, method of inducing specific death of cd38-expressing tumor cells, method of detecting specific death of cd38-expressing tumor cells, method of detecting the presence of cd38 in a minipore source tissue or cell , cd38 detection method in a cd3-expressing erythrocyte 8 and diagnostic composition [P] . BRPI0618399A2 . 2011-08-30

机译：对cd38有特异性的分离的抗原结合区，对cd38有特异性的分离的抗体或其功能片段，分离的抗体或其功能片段，分离的免疫球蛋白，分离的抗原结合区的重链可变区，分离的可变轻链抗原结合区，分离的核酸序列，编码来自分离的抗原结合区的可变重链的核酸序列，核酸序列，载体，分离的细胞，药物组合物，药物组合物的用途，诱导cd38-特异性死亡的方法表达肿瘤细胞的方法，检测表达cd38的肿瘤细胞的特异性死亡的方法，检测小孔源组织或细胞中cd38的存在的方法，表达cd3的红细胞8的cd38检测方法和诊断组合物
5. Control primer annealing to improve the specificity of annealing in nucleic acid amplification kit.Methods for amplifying a nucleic acid sequence of a DNA or a mixture of nucleic acids.And for selectively amplifying a nucleic acid sequence of a target DNA or a mixture of nucleic acids, and a sequence of the target nucleic acid from a mRNA.Method for detecting the complementarity of DNA to mRNA differentially expressed in two or more samples of nucleic acid, methods for amplifying a fragment of the cDNA and DNA quickly target.And to amplify a population of cDNAs of filament double full-length complementary to mRNAs cDNAs, and filament double fortified with 5 'complementary to mRNAs,Method to amplify rather than a sequence of nucleotu00ecdeo target at the same time, methods for producing a digital printing gDNA DNA, and RNA in a sample of mRNA.Method for identifying segments of homology is conserved in a family of multigene from a sample of mRNA, a method for the mutagenesis in a target [P] . BR0214741A . 2004-11-23

机译：控制引物退火，以提高核酸扩增试剂盒中退火的特异性。扩增DNA或核酸混合物的核酸序列的方法以及选择性扩增靶DNA或核酸混合物的核酸序列的方法用于检测DNA与两个或多个核酸样品中差异表达的mRNA的互补性的方法，扩增cDNA和DNA片段的方法可快速靶向。全长与mRNA cDNA互补的双细丝cDNA的cDNA群体，以及与mRNA互补的5'增强双细丝的cDNA，同时扩增而不是核苷酸靶序列的方法，产生数字印刷gDNA DNA的方法mRNA样品中的RNA和RNA.mRNA样品的多基因家族中用于鉴定同源片段的方法是保守的，这是一种在靶标中诱变的方法