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Nucleic acid hybridization chemiluminiscence detection method

A chemiluminescence detection and nucleic acid hybridization technology, applied in the field of nucleic acid hybridization chemiluminescence detection, can solve the problems of high cost of gene sequencing, complicated operation of gene chips, low detection sensitivity, etc., and meet the requirements of fast detection speed, low cost and environmental requirements. low effect

Inactive Publication Date: 2017-08-18
浙江殷欣生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current methods have various deficiencies, such as the high cost of gene sequencing, complex operation of gene chips, low detection sensitivity, and high requirements for laboratories by polymerase chain reaction.

Method used

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  • Nucleic acid hybridization chemiluminiscence detection method

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Embodiment 1

[0032] 1. If figure 1 A nucleic acid hybridization chemiluminescent detection method is shown, which uses a substrate, a capture nucleic acid sequence, a target nucleic acid, a biotin-labeled signal nucleic acid sequence, streptavidin-labeled microspheres and biotin-alkaline phosphatase, The luminescent substrate and the like are used for specific detection of the target nucleic acid to be detected. The specific process is as follows:

[0033] First, according to the sequence AACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGT (SEQ ID No.1) of the target nucleic acid 3; design the capture nucleic acid sequence 2 and modify the amino group: NH 2 -(CH 2 ) 6 - ACCAGAAAGCCACGGCTAACTACG (SEQ ID No. 2);

[0034] Design signal nucleic acid sequence 42, and modify biotin 41,

[0035] Biotin-AACGCTTGCCACCTACGTATTACCGCGC (SEQ ID No. 3);

[0036] The microsphere 51 is specifically a polystyrene microsphere (100 nm in diameter) labeled with streptavidin 52;

[0037] The m...

Embodiment 2

[0055] 1. A nucleic acid hybridization chemiluminescence detection method, according to the sequence AACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGT (SEQ ID No.1) of the target nucleic acid 3;

[0056] Design capture nucleic acid sequence 2 and modify the amino group: NH 2 -(CH 2 ) 6 - ACCAGAAAGCCACGGCTAACTACG (SEQ ID No. 2);

[0057] Design signal nucleic acid sequence 42, and modify biotin Biotin-AACGCTTGCCACCTACGTATTACCGC (SEQID No.3);

[0058] The microsphere 51 is specifically a streptavidin-labeled polystyrene microsphere (100 nm in diameter);

[0059] The matrix 1 is a carboxyl-modified magnetic microsphere with a diameter of 200 nm;

[0060] 2. Take 5 μL (0.1% solid content) carboxyl-modified magnetic microspheres, add 50 μL of 0.1mol / L 2-(N-morpholine) ethanesulfonic acid buffer (MES), 2 μL 0.1mol / L amino-modified Mix the capture nucleic acid sequence, add twice 2.5 μL freshly prepared 10 g / L 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide solution (...

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Abstract

The invention belongs to the technical field of biology, and relates to a nucleic acid detection method, in particular to a nucleic acid hybridization chemiluminiscence detection method. The method comprises the steps that specific detection is performed on target nucleic acid through a matrix, a capture nucleotide sequence, a target nucleic acid, a labeled biotin signal nucleotide sequence, labeled streptavidin microballoons and biotin-alkaline phosphatase, a chemiluminescent substrate and the like; the target nucleic acid is hybridized and captured through the capture nucleotide sequence, signal amplification can be achieved through the signal nucleotide sequence, the microballoons, the biotin-avidin and the alkaline phosphatase activity, so that signals of the target nucleic acid can be detected. Compared with the PCR technology that amplification is needed, the target nucleic acid can be detected on the premise that the concentration of a detection object is not increased, and the method has the advantages of being good in stability, low in cost, high in detection speed, low in environmental requirement and the like.

Description

technical field [0001] The invention relates to a nucleic acid detection method, in particular to a nucleic acid hybridization chemiluminescent detection method, which belongs to the field of biotechnology. Background technique [0002] The current mainstream nucleic acid detection technologies include gene sequencing, gene chip, polymerase chain reaction, etc., as well as technologies based on the amplification of non-target substances, such as the second-generation hybrid capture technology based on antibody capture (HC2 from Kaigen, Germany), Nucleic acid sequence amplification technology based on RNA reverse transcription (Holoje, USA), rapid capture hybridization technology based on branched-chain DNA signal amplification (Cotia Biotechnology), etc. However, all current methods have various deficiencies, such as the high cost of gene sequencing, complex operation of gene chips, low detection sensitivity, and high requirements for laboratories by polymerase chain reactio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/682C12Q2563/103C12Q2563/125C12Q2563/131C12Q2563/149
Inventor 黄宝福章喜超章晓媛叶德王哲
Owner 浙江殷欣生物技术有限公司
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