SNP fluorescence in situ hybridization sequencing detection method for ABCB1 and SLCO1B1

A fluorescence in situ hybridization and detection method technology, applied in the field of molecular genetics, can solve the problems of death, high misjudgment rate, high chip cost, etc., and achieves the effects of high identification, not easy to contaminate, and fast sequencing speed.

Inactive Publication Date: 2019-02-01
北京华夏时代生物工程有限公司
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  • Abstract
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Problems solved by technology

[0012] (3) Adverse reactions between SNP and drugs: In the course of medical treatment, some patients will not only be ineffective after taking the drugs, but some will also have serious adverse reactions, such as improper handling and sometimes death
There are several problems in this type of technology: only one hybridization is performed, and it is difficult to identify non-specific signals, so some non-specific hybridization may cause sequence misreading. In addition, the cost of the chip is high, and the required equipment is expensive, which is not conducive to popularization and application.
This method is prone to errors when the fluorescence curves of homozygous genotypes and heterozygous genotypes are relatively close, and...

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  • SNP fluorescence in situ hybridization sequencing detection method for ABCB1 and SLCO1B1
  • SNP fluorescence in situ hybridization sequencing detection method for ABCB1 and SLCO1B1
  • SNP fluorescence in situ hybridization sequencing detection method for ABCB1 and SLCO1B1

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Embodiment Construction

[0063] The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer along with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention.

[0064] Reaction principle

[0065] Different from ordinary Taqman technology, the present invention does not use PCR technology in the preparation of hybridization templates, but uses single-stranded derivation guide oligonucleotide sequences to convert DNA double-stranded templates into single-stranded derivatives, and Direct hybridization with labeled probes for sequencing.

[0066] The single-stranded derivation requires an oligonucleotide guide sequence capable of complementary binding to the template strand to initiate, and the guide sequence can specifically bind to the annealed template strand at the 3' end and / or 5' end of the template...

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Abstract

The invention discloses a SNP fluorescence in situ hybridization sequencing detection method for ABCB1 and SLCO1B1. The method comprises the following steps: DNA of a sample to be tested is extracted;single-stranded derivatization is performed using the DNA as a template; different fluorescently-labeled first sequencing probes and second sequencing probes are simultaneously added to be hybridizedwith the single-stranded derivative; and finally, the hybridization result is interpreted. The method has high precision and good stability, and is fast, safe and easy for automatic operation. By themethod, precise typing of single nucleotide polymorphism can be completed in the cycle of single-stranded derivatization and hybridization reaction, thereby understanding the genotype of the subjectand achieving prevention of diseases caused by risk factors and clinical guidance for individualized medication.

Description

technical field [0001] The invention discloses a single nucleotide polymorphism detection method, which belongs to the field of molecular genetics. Background technique [0002] With the continuous progress of the development of medical science, many drugs appear in the world every year, but they are eliminated in drug research because of too many adverse reactions. Although the effects and adverse reactions of drugs are related to race and individual differences, genetic differences in genome sequences between people are important reasons for drug efficacy and adverse reactions. Single Nucleotide Polymorphisms (Single Nucleotide Polymorphisms; SNP) is the difference between 1 base between people. There are several genome sequence differences, and SNP is only the most common and common one. Clinically crucial. [0003] SNP is a polymorphism in DNA sequence caused by a single base change. Including single base conversion, inversion and single base insertion or deletion etc...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6841C12Q1/6827C12N15/11
CPCC12Q1/6827C12Q1/6841C12Q1/6886C12Q2600/106C12Q2600/156C12Q2563/107
Inventor 王鹤尧孙美娜
Owner 北京华夏时代生物工程有限公司
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