Nucleic acid detection method based on liquid chip technology

A detection method and liquid chip technology, applied in the field of molecular biology, can solve the problems of time-consuming and high R&D costs, and achieve the effects of reducing the burden of instruments, increasing safety, and saving R&D time and R&D costs.

Pending Publication Date: 2021-12-07
上海万子健生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for different nucleic acids to be tested, it is still necessary to design and synthesize probes that match the nucleic acids to be tested. The project is still time-consuming and the research and development costs are still high.

Method used

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  • Nucleic acid detection method based on liquid chip technology
  • Nucleic acid detection method based on liquid chip technology
  • Nucleic acid detection method based on liquid chip technology

Examples

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Embodiment 1

[0025] This embodiment provides a nucleic acid detection method based on liquid chip technology. This method combines multiplex PCR technology, constant temperature hybridization based on liquid chip technology, detection technology and specific sequence, and the principle of coding microspheres. The typing detection kit for different mutant strains of coronavirus can realize the double-stranded DNA product obtained after PCR after the reverse transcription of the nucleic acid RNA template of the new coronavirus (Bio-Rad reverse transcription reagent is used, see Table 2 for the configuration of the reverse transcription system), And realize the hybridization and signal detection of the probe system under the condition of constant temperature. The detection process includes steps:

[0026] 1) Amplify the pseudoviruses of the mutation sites of the new coronavirus HV69-70del, N501Y and P681H with a labeled primer and a common primer connected to the specific coding Anti-Probe se...

Embodiment 2

[0053] This embodiment provides a nucleic acid detection method based on liquid chip technology. This method combines multiplex PCR technology, constant temperature hybridization based on liquid chip technology, detection technology and specific sequence, and the principle of coding microspheres. The HPV typing detection kit can realize the double-stranded DNA product obtained after the human papillomavirus DNA template is enriched by multiple PCR target sequences, and realize the hybridization and signal detection of the probe system under constant temperature conditions . The detection process includes steps:

[0054] 1) Amplify HPV16, HPV18, and internal reference β-Globin templates with a labeled primer and a common primer connected to a specific coding Anti-Probe sequence (see Table 10 for the specific primer sequence) (the template is synthesized according to NCBI search DNA sequence), the PCR program is as shown in Table 12, and the PCR system preparation is as shown i...

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Abstract

The invention discloses a nucleic acid detection method based on a liquid chip technology. The nucleic acid detection method comprises the following steps: 1) carrying out PCR amplification on a nucleic acid sequence to be detected; 2) preparing a liquid chip; 3) carrying out nucleic acid hybridization; and 5) analyzing a result. A forward primer used in the step (1) is a common primer with a section of specific binding sequence, and the specific binding sequence in the forward primer is defined as an Anti-Probe sequence; and the quantity of the forward primer is equal to that of a reverse primer. The obtained PCR amplification product is a double-stranded DNA product, the double-stranded DNA product is provided with a marker, and the Anti-Probe sequence exists at one end of the double-stranded DNA product in a free single-stranded state. The multi-target detection is realized by adopting multiplex-PCR, and the enrichment of a plurality of nucleic acid target sequences is realized by applying the specific primers. After a specific Anti-Probe binding sequence at the 5'end of an enriched product is combined with Probe which is connected to encoding microspheres and can be complementary with the Anti-Probe binding sequence, the classified detection of a plurality of check table sequences is realized.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a nucleic acid detection method based on liquid chip technology. Background technique [0002] Nucleic acid detection generally requires two processes of PCR amplification and nucleic acid hybridization of the nucleic acid to be tested, and then the product is analyzed. Among them, PCR (Polymerase Chain Reaction) is a molecular biology technique used to replicate and amplify specific DNA fragments in vitro; the principle of PCR is: DNA is denatured to form a single strand at 95°C in vitro; Primers combine with single strands according to the principle of complementary base pairing; DNA polymerase synthesizes complementary strands along the direction from phosphoric acid to five-carbon sugar (5'-3') at ~72°C, so that the product obtained by PCR is usually double-stranded DNA . Nucleic acid hybridization is a process in which complementary nucleotide sequences (DNA and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6837
CPCC12Q1/6837C12Q2531/107C12Q2537/143C12Q2563/149
Inventor 方剑秋沙海天白艳军
Owner 上海万子健生物科技有限公司
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