HER2 gene detection kit and application thereof

Abstract

The invention discloses an HER2 gene detection kit and application thereof and particularly discloses a quantitative HER2 gene detection kit. The kit comprises a PCR reaction buffer solution, a Taq enzyme, a dNTP mixed solution, a standard substance with a gene segment of HER2 mesh and a Fas reference gene segment, a target gene specific primer and a fluorescence probe, a reference gene specific primer and a fluorescence probe and negative control and positive control. A detection method and the detection kit have the advantages that the kit is convenient to use and high in detection efficiency, and the method and the kit have important significance on diagnosis of the HER2 positive breast cancer and research and development, quality control and clinical application of drugs for resistingthe diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a HER2 gene detection kit and its application. Background technique [0002] The human epidermal growth factor receptor-2 (HER2) gene, c-erbB-2 gene, is located on chromosome 17q12-21.32 and encodes a transmembrane receptor-like protein with a relative molecular mass of 185,000. Has tyrosine kinase activity. Under normal human conditions, HER2 is a single-copy gene, but in breast cancer and gastric cancer tumor cells, chromosomal heterogeneity often occurs, such as an increase in the number of chromosome 17, and more commonly, the HER2 gene on chromosome 17 and its The downstream 300-1000M DNA sequence is inserted into other chromosomes through gene rearrangement, which leads to an increase in the copy number of HER2 gene, which in turn leads to the overexpression of HER2 protein on the surface of tumor cells. [0003] Correct detection and evaluation of HER2 pr...

AI Technical Summary

Problems solved by technology

For example, Roche’s HER2 gene amplification detection kit is based on the principle of comparing the copy number difference based on the Ct value difference between the HER2 gene and the reference gene amplification reaction (ΔΔCt after correction of the ΔCt of the reference sample), but different PCR reaction systems There are differences in amplification efficiency and PCR equipment wells, and this method is difficult to meet the stability and precision requirements of clinical HER2 detection

In addition, Thermo Fisher uses a series of target genes and reference genes with different concentration gradients to draw a standard curve, and calculates the copy numbers of the target gene and reference gene in the sample relative to their respective standards, so as to obtain the quantitative copy number of the sample gene, but Due to the high difficulty of absolute quantification of standard products and the large error range of standard product concentration, it is difficult to control the concentration ratio relationship between the target gene and the reference gene, which will cause large errors in the experimental results.

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