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Updated: Nov 15, 2025
Methyl-binding DNA capture Sequencing for Patient Tissues
Published on: October 31, 2016
Detection of free DNA septin 9 gene methylation in plasma
Zhi Luo1, Yingbin Hu2, Xiaoyun Pu3
1Department of Colorectal Surgery, Hunan Cancer Hospital, Changsha 410013, China. 895745492@qq.com.
Objectives:
To explore the correlation between cytosine-phosphoric-guanylic (CpG) site of Septin 9 gene and colorectal cancer, and to develop a real-time PCR detection system in plasma in patients with colorectal cancer.
Methods:
The methylation of training samples was detected by high-throughput sequencing technology, and the sites highly consistent with the clinical information of colorectal cancer were identified. Then the detection system of real-time PCR was designed to analyze the consistency of plasma and tissue based on methylationa sensitive enzyme digestion. Finally, 100 clinical trials were conducted to evaluate the performance of the detection system with the methylation sensitive enzyme digestion-real-time PCR.
Results:
The highly consistent sites, which were selected by high-throughput sequencing from 71 training set samples, was the 38th CpG. Based on the detection region, the screened methylation sensitive enzymes were BstU1, HhaI and HinP1I. The limit for the detection of methylation sensitive enzyme digestion-real-time PCR was 0.5%, and the Ct value was 38.5. The sensitivity was 87.27%, the specificity was 91.49%, the positive predictive value was 92.31%, and the negative predictive value was 86.00%.
Conclusions:
The 38th CpG site of Septin 9 detected by the detection system of methylation sensitive enzyme digestion-real-time PCR can highly predict the occurrence of colorectal cancer with great clinical application value.
