Label-free detection of telomerase activity was done by using telomeric hemin/G-quadruplex triggered polyaniline deposition, not only on themselves but also on the DNA tetrahedron-structure (DTS). DTS size has a great impact on telomerase accessibility, reactivity and detection sensitivity. The method has been used to evaluate bladder cancer development.

Gold nanoparticles (AuNPs) have been extensively explored to be used in analytical methods such as electrochemical, colorimetric methods, and so on. However, only a few methods have been reported by using chirality of AuNPs although their chiral assembly has been studied extensively and circular dichroism (CD) spectroscopy is also a simple and sensitive analytical method. In this paper, sensitive CD spectroscopy method has been explored for detection of 8-hydroxy-2′-deoxyguanosine (8-OHdG), a well-known biomarker for oxidative DNA damage, based on DNA-induced chiroplasmonic assemblies of AuNPs. First, 8-OHdG aptamer hybridized with its complementary sequence that modified with AuNPs based on precision matched bases. DNA-modified AuNPs were assembled into AuNPs dimers by 8-OHdG aptamer, which displayed strong chiroptical activity. Subsequently, in the presence of 8-OHdG, the high specific recognition and affinity constants of aptamer and 8-OHdG destroyed the hybrid of aptamer and its complementary sequence; as a result, AuNPs dimers were destroyed and showed low CD signal. The CD intensity was in log–linear correlation with the concentration of 8-OHdG ranging from 0.05 to 2 nM, with a correlation coefficient of 0.9951 and a detection limit of 33 pM (S/N = 3). The method has been successfully applied in a complex matrix such as human serum samples. The recoveries were from 92.5% to 107% and the relative standard derivations were in the range of 4.89% ∼ 7.27%, indicating that the method had good accuracy and high precision. Therefore, these results indicated that the proposed CD method was simple and reliable, which held great potential for clinical examinations.

Circular dichroism spectroscopy has been explored for detection of methyltransferase activity and inhibition based on DNA-induced chiroplasmonic assemblies of gold nanoparticles and endonuclease HpaII. Good accuracy, precision and sensitivity are obtained in complex matrices such as human serum samples, which is significant for clinical diagnosis and drug development.

•Label-free electrochemical method is used for detection of MTase activity.•HRP-mimicking DNAzyme avoids complex labeling procedures and is robust.•Sensitivity is improved by Exo III and the limit of the detection is 0.12 U mL−1.•The method is applicable to be used in serum samples and is accurate and specific.Detection of DNA methylation and methyltransferase (MTase) activity are important in determining human cancer because aberrant methylation was linked to cancer initiation and progression. In this work, we proposed an electrochemical method for sensitive detection of DNA methylation and MTase activity based on methylation sensitive restriction endonuclease HpaII and the deposition of polyaniline (PANI) catalyzed by HRP-mimicking DNAzyme. In the presence of methylated DNA, HRP-mimicking DNAzyme catalyzed the polymerization of aniline on the dsDNA template, producing huge DPV current. In the presence of non-methylated DNA, dsDNA are cleaved and digested by HpaII and exonuclease III, as a result, no PANI are deposited. This method can be used to determine DNA methylation at the site of CpG. It exhibits a wide linear response toward M.SssI MTase activity in the range of 0.5–0.6 U mL−1 with the detection limit of 0.12 U mL−1. G-rich DNA forms HRP mimicking DNAzyme, which avoids complex labeling procedures and is robust. The method is simple, reliable, sensitive and specific, which has been successfully applied in human serum samples and been used to screen the inhibitors. Thus, the proposed method may be a potential and powerful tool for clinical diagnosis and drug development in the future.

Strategies to detect the methylation of site specific DNA and assay of M.SssI methyltransferase (M.SssI MTase) activity are important in determining human cancers due to aberrant methylation linked to cancer initiation and progression. Herein, we report a label-free fluorescence detection method for DNA methylation and MTase activity based on restriction endonuclease HpaII and exonuclease III (Exo III). A label-free probe DNA was designed, which hybridized with target DNA (one 32-mer DNA from the exon 8 promoter region of the Homo sapiens p53 gene) to form double stranded DNA (dsDNA). Upon the cleavage action of HpaII and degradation reaction of Exo III, dsDNA changed to single stranded DNA (ssDNA) and the fluorescence intensity of thiazole orange (TO) is weak. After the resulting dsDNA was methylated by M.SssI MTase, the action of HpaII and Exo III was prevented, then TO intercalates into the dsDNA and emits strong fluorescence. This method can determine DNA methylation at the site of CpG and distinguish a one-base mismatched target sequence. The fluorescence intensity has a linear relationship with M.SssI MTase activities in the range of 1–10 U mL−1 with a detection limit of 0.16 U mL−1 in terms of 3 times deviation of the blank sample. The methylation of DNA by a hydroxyl radical triggered by DMSO and CH3CHO was also measured. These results show that the proposed method can specifically and selectively detect DNA methylation and M.SssI MTase activity. Human serum has no obvious effects on the assay performance, indicating that the method has great potential for further application in complex samples.

A hairpin molecular beacon tagged with carboxyfluorescein in combination with graphene oxide as a quencher reagent was used to detect the DNA damage by chemical reagents. The fluorescence of molecular beacon was quenched sharply by graphene oxide; while in the presence of its complementary DNA the quenching efficiency decreased because their hybridization prevented the strong adsorbability of molecular beacon on graphene oxide. If the complementary DNA was damaged by a chemical reagent and could not form intact duplex structure with molecular beacon, more molecular beacon would adsorb on graphene oxide increasing the quenching efficiency. Thus, damaged DNA could be detected based on different quenching efficiencies afforded by damaged and intact complementary DNA. The damage effects of chlorpyrifos-methyl and three metabolites of styrene such as mandelieaeids, phenylglyoxylieaeids and epoxystyrene on DNA were studied as models. The method for detection of DNA damage was reliable, rapid and simple compared to the biological methods.Highlights► DNA damage induced by several chemicals was detected by a hairpin MB based on FRET. ► The method was rapid, simple, reliable and sensitive. ► The method can be used to detect damaged DNA induced by other reagents or factors.

Label-free detection of telomerase activity was done by using telomeric hemin/G-quadruplex triggered polyaniline deposition, not only on themselves but also on the DNA tetrahedron-structure (DTS). DTS size has a great impact on telomerase accessibility, reactivity and detection sensitivity. The method has been used to evaluate bladder cancer development.

Circular dichroism spectroscopy has been explored for detection of methyltransferase activity and inhibition based on DNA-induced chiroplasmonic assemblies of gold nanoparticles and endonuclease HpaII. Good accuracy, precision and sensitivity are obtained in complex matrices such as human serum samples, which is significant for clinical diagnosis and drug development.