Method for performing in-vitro accurate test on KRAS gene mutation

Abstract

The invention provides a method for performing in-vitro accurate test on KRAS gene mutation, and the method is used for overcoming the defects that the conventional allele-specific polymerase chain reaction (PCR) method has insufficient specificity and false positive phenomenon. The method for performing the in-vitro accurate test on the KRAS gene mutation comprises the following steps of: (1) designing and synthetizing allele specific primers which comprise the allele specific primers espectively designed for 7 types of mutation of a typical KRAS gene exon 2 and are utilized as forward primers, wherein extension retardant primers are utilized as common downstream primers; (2) extracting the KRAS genomic deoxyribose nucleic acid (DNA) of a test sample, and additionally preparing the wild type genomic DNA (as wild type template); and (3) carrying out the real-time fluorescent PCR reaction. The test method has the advantages of strong specificity, high sensitivity, material conservation and time-saving property.

Description

technical field [0001] The invention relates to a method for detecting KRAS gene mutation. Background technique [0002] Gene mutation refers to the change of base composition or arrangement sequence in the structure and function of genomic DNA molecules, mainly including base substitution and fragment insertion and deletion, and is one of the important causes of genotype diseases. Gene mutation analysis plays a very important role in biomedical research, especially in genotypic disease diagnosis and pathological research. The protein encoded by the KRAS gene is involved in the cell signal transduction pathway mediated by the epidermal growth factor receptor (EGFR), affecting cell proliferation, growth and metastasis; the protein encoded by the normal KRAS gene is activated after receiving upstream signal activation, and The inactivated state will be restored after the signal is transmitted to the downstream cytokines; while the protein encoded by the mutated KRAS gene is a...

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the shortcomings of the traditional allele-specific PCR method, such as insufficient specificity and false positive phenomenon, the present invention provides a method for accurately detecting KRAS gene mutations in vitro. Accurate detection of KRAS mutation in vitro by PCR method

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