Real-time fluorescent quantitative PCR detection method of human breast cancer genotype

Abstract

The invention discloses a real-time fluorescent quantitative PCR detection method of human breast cancer genotypes. The method comprises the following steps: ERa (SEQID9), PR(SEQID10) and HER-2(SEQID11) of the human breast cancer gene are taken as augmented target gene sequences, 18SrRNA(SEQID12) is taken as an internal control gene, specific primers are designed corresponding to the ERa gene, the PR gene, the HER-2 gene and the 18SrRNA gene respectively, CT values of a sample to be detected and a control sample are detected through adopting the real-time fluorescent quantitative PCR method, and the target gene expression level is calculated; the target gene expression level is analyzed through the delta delta CT method, the target gene expression level equals to 2<delta delta CT>, wherein delta delta CT equals to (CT, target minus CT, 18S)(detected) minus (CT, target minus CT, 18S)(control); when the target gene expression level is less than or equal to 1, the ERa gene, the PR gene and the HER-2 gene are subject to negative typing; and when the target gene expression level is more than 1, the ERa gene, the PR gene and the HER-2 gene are subject to positive typing. The invention has the advantages of simplicity and convenience in operation, high detection sensitivity, short detection time, good specificity and repetitiveness, accurate and reliable results and qualitative and quantitative detection, can be used for genotype identification of common populations or patients with breast cancer, provides an important theoretical index on personalized for the patients with breast cancer to personally take medicine, has significance for prognosis of the patients with breast cancer, and has stronger popularization and practicability.

Description

technical field [0001] The invention belongs to the field of physiological and biochemical detection, in particular to a real-time fluorescent quantitative PCR detection method for genotyping of human breast cancer. Background technique [0002] Breast cancer is one of the main malignant tumors that endanger women's health. About one million women in the world suffer from breast cancer every year, and about 500,000 women die from the disease. In recent years, the incidence of breast cancer in China has shown an obvious upward trend, especially in economically developed coastal areas, where the incidence of breast cancer has ranked first among female malignant tumors. The prognosis of breast cancer is closely related to the invasion and early metastasis of cancer cells, and metastasis is the main cause of death in breast cancer patients. How to screen patients with potential metastasis risks early and give targeted treatment is the key to improve the survival rate of breast ...

AI Technical Summary

Problems solved by technology

[0010] The present invention overcomes the shortcomings of conventional PCR that cannot be quantified and may have non-specific amplification in the prior art, and proposes a real-time fluorescence quantitative PCR detection method for human breast cancer genotyping, which has both high efficiency of DNA amplification, Beneficial effects of high specificity of probe technology and high sensitivity of spectroscopic technology

Images