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

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...

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.

[0025] In view of the technical problems such as high misjudgment rate, poor accuracy, low stability, and low detection efficiency in the SNP detection of the membrane transporter encoding genes ABCB1 and SLCO1B1 in the prior art, the present invention intends to provide an improvement on the prior art. , by designing single-stranded derived guide sequences and probe sequences, optimizing each component, providing a more accurate, stable, and faster SNP fluorescence in situ hybridization sequencing of the membrane transporter-encoded genes ABCB1 and SLCO1B1 Detection methods to provide guidance for clinical genetic testing and individualized medicine

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