Primer pair sets and kits for detection of genetic modification differences in cytosine deaminase and related molecules in cfDNA

Abstract

The invention discloses a primer pair group and a kit for detecting differences in gene modification of cytosine deaminase and related molecules in cfDNA. The present invention firstly discloses a set of primer pairs for detecting differences in epigenetic modification of cytosine deaminase and related molecular genes in free peripheral blood DNA. The primer pairs consist of SEQ ID No.1 to SEQ ID No. . The nucleotide sequence composition shown in 15. The present invention further discloses a kit comprising the above primer set. The present invention is a primer set and a kit for detecting differences in epigenetic modification of cytosine deaminase and related molecular genes in free DNA in peripheral blood. The detection has high sensitivity and specificity, and is useful for tumor risk prediction and efficacy evaluation important value.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, it relates to a primer pair set and a kit for detecting differences in genetic modification of cytosine deaminase and related molecules in cfDNA. Background technique [0002] 1. Liquid biopsy: In the clinical needs of tumor prevention and treatment, liquid biopsy has become an important means of tumor risk prediction, targeted drug guidance and efficacy evaluation, among which early tumor risk assessment is the most important link in tumor prevention. Blood is undoubtedly the best sample for liquid testing because it contains a large amount of tumor-related information, among which free DNA in peripheral blood is currently the focus of liquid biopsy. This free DNA exists in plasma or serum, and it may come from the release of normal cell death. It may also come from the death release of tumor or precancerous cells, and both normal cells and tumor cells can be actively rel...

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Problems solved by technology

[0003] 2. Detection of structural changes: Since tumor cells involve different degrees of gene mutations, the detection of structural changes has become a focus of attention. The detection of this target is relatively easy in tumor solid tissues, because in solid tumor tissues, The abundance of target genes containing mutations is high, but it is more difficult in peripheral blood, because the concentration of DNA (ctDNA) from tumor cells in peripheral blood is very low, and mutations at some sites in target genes The frequency is also very low. In addition, it should be emphasized that the location and frequency of structural changes are very different in many tumors, and they are random, and these structural changes are still a process of continuous accumulation during the course of the disease. Therefore, the superposition of these factors will lead to a significant decrease in the detection sensitivity

It is known that the degree of methylation of gene promoters is negatively correlated with gene expression (affecting the binding of transcription factors and RNA polymerase), but the function of epigenetic modifications in gene bodies is still unclear. Current research mainly focuses on Epigenetic modifications to gene promoter regions, while little attention has been paid to changes in various epigenetic modifications in the gene body

[0006] 4. Selection of target genes: The identification of tumor-related target genes has long been a hotspot in tumor molecular detection. The target genes identified based on samples at a certain time point in the occurrence and development of tumors only represent a section of the tumor gene map. It also includes some transient changes, which cannot reflect its position and dynamic changes in the process of tumorigenesis and development

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