A method is described for the colorimetric determination of the activity of CpG methyltransferase (M.SssI). It is based on (a) the crosslinking effect between dsDNA-modified gold nanoparticles (AuNPs) and graphene oxide (GO), and (b) an amplification reaction with the aid of a nicking enzyme. To avoid the aggregation of AuNPs (which would produce false signals), a hairpin DNA was connected to the AuNPs. Thus, the red color of the solution (measured at 530 nm) increases linearly with the activity of M.SssI from 0.2 to 60 U·mL−1, and the limit of detection is 67 U·mL−1. This is superior to some reported strategies. The method was successfully applied to analyze spiked serum samples. Conceivably, it represents a powerful tool for use in drug development and diagnosis.
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This work is supported by the National Natural Science Foundation of China (21605075; 21365015) and Natural Science Foundation of Jiang Xi Province 20181BAB203020.
Human serum sample used in this work was obtained from a healthy volunteer. This study was performed on the consent of the volunteer and in accordance with the ethical standards of the Medical Ethics Committee of the First Affiliated Hospital of Nanchang University (Jiangxi, China), and with the 1964 Helsinki declaration on Biomedical Research Involving Human Subjects and its later amendments. The authors declare that they have no competing interests.
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Li, ZM., Zhang, X., Pi, T. et al. Colorimetric determination of the activity of methyltransferase based on nicking enzyme amplification and the use of gold nanoparticles conjugated to graphene oxide. Microchim Acta 186, 594 (2019). https://doi.org/10.1007/s00604-019-3690-4
- Crosslinking effect
- M. SssI activity
- Hairpin DNA
- Drug development