Method of combined detection for diagnosis of breast cancer by adopting ddPCR technology

Abstract

The invention discloses a method of combined detection for diagnosis of a breast cancer by adopting a ddPCR technology. The method specifically comprises the following steps of S1, extracting genomicDNA and miRNA from a sample; S2, carrying out modification and purification quantification through DNA sulfite and reversely transcribing the miRNA into cDNA; S3, separately carrying out ddPCR by employing purified DNA genome as a template and a specific primer as a primer for detecting a methylation state of MASPIN; and S4, calculating methylation degree W of an upstream regulatory region CpG island of a maspin coding sequence of a to-be-detected sample and combining the methylation degree W with the content of miR-142 in the sample for analysis. The invention relates to the technical field of biology and disease diagnosis. According to the method of combined detection for diagnosis of the breast cancer by adopting the ddPCR technology, combined detection of two indicators, namely the methylation state of the maspin gene and the content of the miR-142 can be used for evaluating whether a patient suffers from the breast cancer or not, the data are more accurate, the result is more reliable, and the method can be widely applied to early screening for the breast cancer and effectively assists judgement the malignant degree and metastasis of the breast cancer.

Description

technical field [0001] The invention relates to the technical field of biology and disease diagnosis, in particular to a method for diagnosing breast cancer by using ddPCR technology combined with methylation-specific PCR to detect gene maspin methylation status and miR-142 relative content. Background technique [0002] Currently, the most commonly used imaging methods for breast cancer screening are mammography and ultrasound. Mammography, the full name of mammography X-ray examination, also known as palladium examination, is the first choice and the easiest and most reliable non-invasive detection method for the diagnosis of breast diseases. High quality, good repeatability, and the obtained images can be compared before and after, and are not limited by age and body shape, and are currently used as routine inspections. It is characterized by the detection of breast lumps that cannot be touched by doctors, especially for large breasts and fatty breasts, and its diagnosti...

AI Technical Summary

Problems solved by technology

[0003] (1) Cannot detect the occurrence of early breast cancer in situ, that is, it is difficult to detect when no obvious tumor is formed or the tumor mass is small in the early stage of breast cancer

[0004] (2) It is difficult to predict whether breast cancer will metastasize

[0005] (3) It is difficult to detect early metastasis of breast cancer

[0006] In short, the existing detection technology can no longer meet the requirements of clinicians for preoperative histopathological grading and prediction of tumor molecular typing in tumor patients.

Images