The EpiTect Methyl qPCR Assay as novel age estimation method in forensic biology

Abstract

Human aging is associated with epigenetic modification of the genome. DNA methylation at cytosines appears currently as the best characterised modification that occurs during the mammalian lifetime. Such methylation changes at regulatory region can provide insights to track contributor age for criminal investigation. The EpiTect Methyl II PCR system (QIAGEN) was used to compare methylation levels of CpG islands in the promoter regions of a number of age related genes, of which four successfully showed changes across the lifespan (NPTX2, KCNQ1DN, GRIA2 and TRIM58). This technique is based on the detection of remaining input genome after digestion with a methylation-sensitive restriction enzyme. This study examined DNA specimens from 80 female subjects of various ages (18-91 years) obtained from blood, using primers designed to flank the studied gene loci. The data obtained from DNA methylation quantification showed successful discrimination among volunteered ages. Overall, the difference between predicted and real age was about 11 years and absolute mean differences (AMD) was only 7.2 years error. We suggest the EpiTect system can be used as fast and simple innovative tool in future forensic age estimation.

Investigating the expression of MMPs and TIMPs in preterm placenta and role of CpG methylation in regulating MMP‐9 expression

“…MMP‐1 is the key enzyme that participates in ECM turnover and MMP‐2,‐9 digest the major components of basement membrane . Our results are consistent with previous studies showing increased placental MMP‐1 expression during labor . MMP‐9 expression is up regulated during labor , both at term and preterm, and increased activity of MMP‐9 has been associated with preterm birth .This data further reiterates the role of MMPs as mediators in placental ECM remodeling associated with term and preterm labor.…”

“…In this study, CpG methylation in promoter region of MMP‐9 gene was examined using the EpiTect Methyl qPCR Assay for gene methylation analysis. The results of this assay have been reported to be comparable to that of bisulfite Sanger sequencing and Infinium BeadChip experiments and this method has also been employed in several other studies for determination of gene promoter methylation . Our data indicates lower CpG methylation in the promoter region of MMP‐9 gene in the preterm placenta as compared to term.…”

DNA methylation-based forensic age prediction using artificial neural networks and next generation sequencing

“…Also, while most of these studies are based on targeted methylation detection via pyrosequencing, qPCR [36], melting curve analysis [37] and the EpiTYPER system [38], [39] have also been used. Although pyrosequencing has been the gold standard for such analysis since its introduction [40], [41] and shows various advantages over other methylation techniques [42], it is mainly performed as single reactions because multiplex pyrosequencing can be complex [43].…”

Estimation de l’âge médicolégal grâce à l’étude de la méthylation de l’ADN : revue de la littérature

“…MSRE-PCR a été décrit dans le passé comme un moyen rapide et fiable de détecter simultanément les profils de méthylation de l'ADN dans plusieurs fragments, de manière sensible et reproductible [16]. L'équipe de Mawlood [17] a utilisé la technique Epitect Methyl qPCR system (QIAGEN) pour prédire un âge dans une cohorte de 80 femmes âgées de 19 à 91 ans à partir d'échantillons sanguins. Il a comparé les niveaux de méthylation des dinucléotides CpG dans les régions promotrices de quatre gènes liés à l'âge (NPTX2, KCNQ1DN, GRIA2 et TRIM58).…”