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DNA Methylation Detection and Tumors

  • Time of issue:2021-10-29 08:59

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DNA Methylation Detection and Tumors

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  • Time of issue:2021-10-29 08:59
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DNA Methylation Detection and Tumors

Biological Basis of ctDNA Detection Technology

Circulating tumor DNA (ctDNA) is a type of circulating cell-free DNA (cfDNA). As a research focus in the field of oncology detection in recent years, it has an important application value in the non-invasive early screening of tumors.

Figure 2. Features of ctDNA

 

Clinical Application of DNA Methylation

The high concordance of abnormal methylation of ctDNA in specific genetic regions makes DNA methylation detection more widely used than detection of tumor-specific mutations in tumor diagnosis, surveillance, treatment efficacy and prognosis determination. Therefore, ctDNA methylation detection is considered one of the most valuable methods for cancer diagnosis and risk assessment.

The high concordance of abnormal methylation of ctDNA in specific genetic regions makes DNA methylation detection more widely used than detection of tumor-specific mutations in tumor diagnosis, surveillance, treatment efficacy and prognosis determination. Therefore, ctDNA methylation detection is considered one of the most valuable methods for cancer diagnosis and risk assessment.

Compared to traditional tumor biopsy techniques, ctDNA liquid biopsy has more advantages, such as faster sampling, lower price, less invasiveness, and the ability to track a range of tumor developments.

 

Prediction of Prognosis Risk

Abnormal DNA methylation can indicate tumor aggressiveness and risk of recurrence after surgical resection. As ctDNA has a short half-life (about 16 min~2.5 h), and some studies have shown that the persistence of ctDNA in the blood after surgery is associated with poor prognosis, the methylation profile of ctDNA can help predict the risk of postoperative recurrence and death in cancer patients and help to adjust the therapeutic regimen, assess the need for postoperative chemotherapy and determine the chemotherapy regimen.

 

Evaluation of Therapeutic Efficacy

Body fluid biopsy collection of ctDNA can be repeated multiple times, which is very beneficial in assessing the efficacy of treatment and monitoring the patient's physical condition in real time during the course of disease. In contrast to imaging or traditional protein-based marker assays, ctDNA levels and their trends can indicate a patient's response to therapeutic agents. Body fluid biopsies can also be used to monitor patients receiving targeted drug therapy, suggesting risk of relapse and drug resistance-associated mutations.

 

Common Detection Methods of DNA Methylation

Many techniques for DNA methylation detection have emerged in recent years, and can be classified into three categories based on the means of pre-treatment of the target DNA.

Methylation Detection Based on Restriction Enzyme Pre-treatment

This method utilizes the property of methylation-sensitive restriction endonucleases (MSRE) that do not cleave the methylated region to digest DNA into fragments of different sizes before analysis. The commonly used enzyme pair is Hpa II - Msp I (recognition sequence CCGG). Then Southern or PCR amplification of the isolated product is performed to clarify the methylation status of the target fragment.

Figure 4. MSRE Method

 Methylation Detection Based on Sulfite Modification Pre-treatment

In this method, DNA is treated with sulfite and unmethylated cytosine (C) is converted to uracil (U), while methylated cytosine remains unchanged. Through sequencing or PCR with specific primers, accurate information on the methylation sites of the DNA sequence is obtained.

 

Methylation Detection Based on Affinity Enrichment Pre-treatment

This method mainly includes methylated DNA immunoprecipitation (MeDIP) and methylated CpG-binding protein affinity capture (MBDCap): the MeDIP method enriches methylated fragments using 5-methylcytosine-specific antibodies after ultrasonically interrupting and denaturing genomic DNA, and then isolates and purifies them to obtain methylated DNA fragments, which are then then analyzed by sequencing and other methods; MBDCap technique is similar to the MeDIP method, using methylated DNA binding proteins to immunoprecipitate methylated DNA and, as the enrichment proteins used are different, MeDIP generally enriches methylated regions with low density of CpG, while MBDCap generally enriches methylated regions with high density of CpG.

Figure 6. MeDIP

 

Product Recommendation

Based on DNA methylation detection, Vazyme offers molecular diagnostic materials and solutions. We can provide qPCR products with high specificity to meet the requirements of DNA methylation detection. We can also provide total solutions and customized services.

 

[1] Kaneko M, Kotake M, Bando H, et al. Prognostic and predictive significance of long interspersed nucleotide element-1 methylation in advanced-stage colorectal cancer[J]. Bmc Cancer, 2016, 16(1):945.

[2] Sun K, Jiang P, Chan K C A , et al. Plasma DNA tissue mapping by genome-wide methylation sequencing for noninvasive prenatal, cancer, and transplantation assessments[J]. Proceedings of the National Academy of Sciences, 2015, 112(40).

[3] [1]Chetan, Bettegowda, Mark, et al. Detection of circulating tumor DNA in early and late stage human malignancies.[J]. Neuro Oncology, 2014.

 

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