Real-time fluorescence quantitative PCR (Polymerase Chain Reaction) kit for detecting expression level of HER2 genes

Abstract

The invention provides a real-time fluorescence quantitative PCR (Polymerase Chain Reaction) kit for detecting the expression level of HER2 genes and relates to a PCR kit. The real-time fluorescence quantitative PCR kit comprises PCR reaction liquid of the HER2 genes, PCR reaction liquid of ACTB genes, a Taq enzyme system, a control product and a packaging object; and the adopted primer is an annular primer, 3-8 self-designed basic groups are at the 5' end, and can be combined with the sequence at the 3' end under proper conditions so as to form double chains. The real-time fluorescence quantitative PCR kit has the advantages that the non-specific amplification products especially primer dimer generated by nonspecific amplification products especially the primer can be effectively prevented from being formed, the specificity is increased; and a relative-quantitative RT-PCR method is utilized for detecting the expression level of the HER2 genes of a patient with the breast cancer, adopts a 2-delta deltaCt value method for quantifying the detected result, and can be used for diagnosis of early stage and transferring of the breast cancer and assisting multiple fields such as clinicalmedicine selection and prognosis.

Description

technical field [0001] The invention relates to a PCR kit, in particular to a real-time fluorescent quantitative PCR kit for detecting the expression level of HER2 gene. Background technique [0002] Breast cancer is the second most common tumor in women, second only to non-melanoma skin cancer, and the second leading cause of death among women with cancer. The incidence is often related to heredity. According to epidemiological surveys, 5% to 10% of breast cancers are familial. Since the 20th century, the incidence of breast cancer has been increasing all over the world. According to statistics, the number of deaths from breast cancer in the world in 2008 alone was as high as 458,503, accounting for 13.7% of female malignant tumor deaths (Boyle P, Levin B. World Cancer Report 2008 [M]. France: IARC, 2008). In recent years, the incidence of female breast cancer in my country is also rising sharply at an annual growth rate of 3% to 4%. About 200,000 women suffer from breast...

AI Technical Summary

Problems solved by technology

[0008] Immunohistochemical detection of HER2 protein expression is currently the most commonly used method. It uses a specific antibody labeled with a chromogenic agent to qualitatively and A new technology for localization and quantitative determination, IHC method has been widely used in clinical pathological diagnosis due to its advantages of simple operation, low price, convenient interpretation, and easy storage, but due to the lack of standardization of the detection process, and the results are easily affected by tissue processing , the difference in sensitivity of different antibodies, the difference in subjective judgment of observers, and the influence of the interference of chromosome 17 polysomy often cause the difference in the test results

[0009] In comparison, the fluorescence in situ hybridization method hybridizes a fluorescently labeled DNA probe to a DNA target sequence in the nucleus, and uses a fluorescent detection system to perform qualitative, quantitative or relative positioning analysis of the DNA to be tested, which detects frozen tissue or formalin The stability of HER2 gene amplification in fixed and paraffin-embedded tissues is better, and it is not easily affected by tissue processing, etc., and the HER2 gene and chromosome 17 centromere are simultaneously marked with different colors of fluorescent signals, eliminating chromosome 17 polysomy However, FISH also has disadvantages such as expensive detection cost, long-term preservation of results due to fluorescence quenching, difficult operation, long time, and it is not conducive for pathologists to observe the morphology of tissue cells under dark field, which leads to false positive or false positive results of HER2. Negative misjudgment and other defects

Images