Method for detecting methylation of SHOX2 gene in lung tumor tissue DNA

Abstract

The invention discloses a method for detecting methylation of an SHOX2 gene in lung tumor tissue DNA. The method comprises the following steps: 1) sulfite modification treatment is performed on a to-be-detected lung tumor tissue DNA sample; 2) the sulfite-treated lung tumor tissue DNA, 20%, 40%, 60%, 80% and 100% methylation standard DNA, an SHOX2 specific PCR detection primer, an SHOX2 specific probe, an internal reference gene primer and an internal reference gene probe undergo a quantitative reverse transcription-polymerase chain reaction (qRT-PCR) reaction; and 3) Ct values of the lung tumor tissue DNA and the 20%, 40%, 60%, 80% and 100% methylation standard DNA in the SHOX2 gene are measured. According to the method for detecting the methylation of the SHOX2 gene in the lung tumor tissue DNA, the methylation state of the SHOX2 gene can be accurately and effectively distinguished, the specific primer is reasonably selected, and thus, the sensitivity and specificity of detection canbe further improved.

Description

technical field [0001] The invention relates to scientific research and clinical fields that need to detect the methylation of the SHOX2 gene in the DNA of lung tumor tissues, and specifically relates to a method for detecting the methylation of the SHOX2 gene in the DNA of lung tumor tissues. Background technique [0002] DNA methylation is a common epigenetic modification in the human body, which can regulate the expression of modified genes, so it is considered to be closely related to the occurrence of tumors. Early screening of tumors, auxiliary diagnosis, treatment, and prognosis can be completed by detecting the methylation status of tumor-related genes. At present, the detection of DNA methylation status mainly uses sulfite pretreatment to convert unmethylated cytosine (C) in DNA samples into uracil (U), while methylated cytosine Pyrimidine (C) cannot be converted and remains in its original state, and then the methylation status of its related genes is detected by ...

AI Technical Summary

Problems solved by technology

However, the sensitivity and specificity of MSP are poor, and it is impossible to detect small changes in DNA methylation status, which hinders its application in the regulation of methylation of tumor-related genes.

In addition, the methylation fluorescence detection technology based on the PCR method, such as the patent CN103732759, CN106480210A, although the sensitivity of the detection has been improved, but both have the disadvantages of cumbersome operation, high cost, and long detection cycle

Images