Nucleotide molecule combination for fluorescence in situ hybridization sequencing detection of SNP and application thereof

Abstract

The invention discloses a nucleotide molecule combination for fluorescence in situ hybridization sequencing detection of SNP and an application method thereof in detecting SNPs in tumor drug treatmentrelated genes UGT1A1, DPYD and XRCC1. 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 probe and second sequencing probe are simultaneously added to be hybridized with the single-stranded derivative; and finally, the hybridization result isinterpreted. The method has high precision and good stability, and is fast, safe and easy for automatic operation. By the method, 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 subject and 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 Significant. [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...

AI Technical Summary

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 cannot give accurate results, which may easily cause false positives or misjudgments.

[0028] In view of the technical problems such as high misjudgment rate, poor accuracy, low stability, and low detection efficiency in the detection of SNPs related to tumor occurrence and treatment such as UGT1A1, DPYD, and XRCC1 in the prior art, the present invention intends to provide a new solution to the existing SNPs. The technology is improved, and by designing single-stranded derivative guide sequences and probe sequences, optimizing each component, a SNP hybrid sequencing method with higher accuracy, stability and speed is provided for clinical gene detection and individualization Guidance on medication

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