DNA methylation test is divided into two major steps. First, a chemical called Bisulfite is used to distinguish methylated and unmethylated cytosine. The unmethylated cytosine converted to Uracil by Bisulfite, and methylated cytosine (C-CH3) remains as cytosine. The methylation status is then identified by the methylation-specific PCR method or base sequencing analysis called Pyrosequencing. Unmethylated DNA is not amplified due to the genetic modification by Bisulfite, but DNA methylated genes are amplified.
It is difficult to obtain accurate results in DNA methylation test using Bisulfite because a lot of tissue or blood should be collected from the patient. it requires a lot of DNA, and the rate of false-negative is high due to non-specific genetic variation by Bisulfite. Also, as its low reproducibility, it takes a long time to repeat tests (more than 3 times) so it can be hard to apply to accurate and rapid molecular diagnosis.
Epi-TOP technology is the world’s first breakthrough technology that enables to amplify genes quickly without having to process samples in advance using chemicals such as bisulfite, which has been required for DNA methylation testing. This technology also integrates the Real-Time PCR, a widely used molecular diagnostic tool, to analyze the methylation status easily and accurately. Epi-TOP can selectively combine its own "Epi-sPNA" probe to target genes to suppress gene amplification, and only methylated genes are selected for amplification. This allows us to evaluate the ratio of methylated genes by comparing the ratio of Ct and Tm values.
Epi-TOP Technical Advantages - Fast: Finished within three hours
- Reproducibility : Results are derived by a single experiment
- Convenience :
Sample pre-processing, such as Bisulfite, is not necessary
- Sensitivity: Methylation level is stably measured up to 0.05%
- Specificity: No cross-reaction to non-methylated genes
The in vitro diagnostic medical device that helps in molecular genetic differential diagnosis by qualitatively determining whether MGMT is methylated in glioblastoma patients
The in vitro diagnostic medical device that aids in the diagnosis of Lynch syndrome and MLH1 promoter methylation in patients with sporadic tumors caused by MMR deficiency
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