Fluorescent polarization based homogeneous phase detection method of single nucleotide polymorphism of codon118 of ERCC1 (excision repair cross-complementing 1) gene

Abstract

The invention relates to a fluorescent polarization based homogeneous phase detection method of a single nucleotide polymorphism of a codon118 of an ERCC1 (excision repair cross-complementing 1) gene. The homogeneous phase detection method solves the problems of high cost, complicated operating steps, proneness to causing contamination and influence on detection result accuracy in the prior art, and is simple in steps, easy to operate, not prone to causing contamination as operation can be completed once in a closed tube, and low in cost as any special reagent and fluorescence quenching or micro-groove binding agent are not needed. Application of a specific probe with single end marked to detection of the single nucleotide polymorphism of the codon118 of the ERCC1 gene results in changes of fluorescent polarization values and enables detection results to be more objective and accurate. Result analysis is simple as only numeric comparison is needed in result judgment, and the homogeneous phase detection method can easily realize standardization and automation, is wide in application range and can be used for detecting clinical blood or tissue specimens.

Description

Technical field: [0001] The present invention relates to the technical field of detection of Codon118 single nucleotide polymorphism at the 118th codon of ERCC1 gene Exon4, in particular to a homogeneous detection method, which monitors the presence of ERCC1 gene Exon4 in a sample by detecting the changed fluorescence polarization value Codon118 single nucleotide polymorphism in codon 118. Background technique : [0002] Single nucleotide polymorphism (SNP) refers to the presence of different bases at specific nucleic acid positions in the genome of normal individuals of a certain population, which is the difference in human DNA sequence, that is, when a single nucleotide in the genome sequence of the chromosome changes. Polymorphic changes in DNA sequences, SNPs are also known as biallelic markers. Among the 3 billion bases in the human genome, about one SNP occurs every 300 to 500 bases, and there are about 20 million SNPs in the entire human genome. SNP can occur in bo...

AI Technical Summary

Problems solved by technology

[0004] At present, there are many analysis methods for Codon118 single nucleotide polymorphism in codon 118 of ERCC1 gene, such as RFLP, allele-specific oligonucleotide hybridization, Oligonucleotide ligation analysis, allele-specific PCR, DNA sequencing, gene chip, TDI-FP and other technologies can be detected separately, but these methods involve steps such as amplification, digestion and purification, inactivation, hybridization extension and pipetting, etc. More electrophoresis and multi-step post-processing are required, which is time-consuming and labor-intensive, and the efficiency needs to be improved

Moreover, during the operation process, the reaction tube needs to be replaced, which is prone to contamination, which affects the accuracy of the test results.

Or the instruments and equipment are expensive, and it is difficult to promote and apply them on a large scale

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