Non-amplification nucleic acid hybrid capture system and application thereof in nucleic acid test

A nucleic acid hybridization and non-amplification technology, which is applied in the non-amplification nucleic acid hybridization capture detection system and its application in nucleic acid detection, can solve the problems of high detection cost, complicated operation, and inability to truly reflect nucleic acid. achieve low cost

Inactive Publication Date: 2017-12-29
NANJING UNIV
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Problems solved by technology

[0005] Problems to be solved in the present invention: In view of the shortcomings of existing nucleic acid detection methods, such as relatively complicated operation, corresponding amplification and detection instruments are required, resulting in high detection costs, which cannot truly reflect the initial amount of nucleic acid

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  • Non-amplification nucleic acid hybrid capture system and application thereof in nucleic acid test
  • Non-amplification nucleic acid hybrid capture system and application thereof in nucleic acid test
  • Non-amplification nucleic acid hybrid capture system and application thereof in nucleic acid test

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Embodiment Construction

[0038] Specific examples are provided below to further illustrate the technical solution of the present invention, but the application of the technology of the present invention is not limited to the examples.

[0039] 1 Extraction of HPV16 RNA

[0040] Extract RNA from HPV16 positive or negative samples.

[0041] 2 Design combinations for HPV16 RNA oligonucleotide probes

[0042] According to the HPV16 genome sequence published by NCBI (reference sequence: NC_001526.2), the biotin-labeled oligonucleotide probes for each gene fragment of HPV16 are designed, and the sequence is as shown in Table 1:

[0043] Table 1 Probe sequence

[0044]

[0045] 3 Establishment of immunological detection method for HPV16 RNA

[0046] 3.1 Annealing hybridization

[0047] Anneal and hybridize the above probe system with HPV16 RNA template, hybridization system: 10 μL 10× annealing buffer, 13 μL or 12 μL probe mixture, 10 μL RNA template solution, supplemented with DEPC H 2 0 to 100 μL; ...

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Abstract

The invention belongs to the technical field of molecular biology, immunology and nucleic acid chemistry and in particular relates to a non-amplification nucleic acid hybrid capture system and application thereof. The kit is an immunoassay method for capturing hybrid nucleic acids based on a non-amplification anti-nucleic antibody; inverse transcription, PCR (Polymerase Chain Reaction) and other amplification technologies are not needed in the detection process, while a probe complementary to a to-be-detected nucleic acid is subjected to annealing hybridization with the to-be-detected nucleic acid so as to form a structure which can be recognized by the anti-nucleic antibody, and the detection signal is amplified by utilizing enzyme linked immunosorbent assay. The detection method has the advantages of being rapid, low in cost, high in sensitivity, high in specificity, free in complicated amplification and detection instruments and capable of detecting initial viral load.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology, immunology and nucleic acid chemistry. In particular, it relates to a non-amplified nucleic acid hybrid capture detection system and its application in nucleic acid detection. Background technique [0002] Nucleic acid is a biological macromolecular compound synthesized by many nucleotides, and is one of the most basic substances of life. Nucleic acid widely exists in all animal and plant cells and microorganisms. Different nucleic acids have different chemical compositions and nucleotide arrangement sequences. According to different chemical composition, nucleic acid can be divided into ribonucleic acid (referred to as RNA) and deoxyribonucleic acid (referred to as DNA). [0003] Currently, nucleic acid detection methods are mainly divided into DNA detection and RNA detection. For DNA detection, there are methods based on in situ hybridization and DNA sequencing, based on branched...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/70
CPCC12Q1/6804C12Q1/708C12Q2565/518C12Q2565/519C12Q2527/125
Inventor 沈萍萍丁森黄亚红刘培
Owner NANJING UNIV
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