Amplification strategy for sensitive detection of methyltransferase activity based on surface plasma resonance techniques.

In this work, a newly developed surface plasma resonance (SPR) system for the sensitive detection of M.SssI activity has been designed based on double signal amplification with DNA chain cyclic reactions and AuNPs. In the absence of M.SssI, hairpin DNA 1 (HP1) can be cleaved into s1 fragments catalyzed by HpaII. The s1 fragments can then trigger a recycling process of hairpin DNA 2 (HP2) hybridization and subsequently release massive s2 and s3 in the solution of Nt.AlwI and HPII. AuNPs-DNA can be captured on gold film by the released s2 and s3 to produce a strong SPR signal. Whereas in the presence of M.SssI, methylated HP1 cannot be cleaved by HpaII, thus produce a weak SPR signal. The SPR signals are dependent on the M.SssI concentration in the range from 0.5 to 50 U/mL. The successful detection of M.SssI activity in clinical serum samples and inhibition of M.SssI using 5-Aza and 5-Aza-dC indicate a great potential of this strategy for building new monitoring platform in bioanalysis and clinical biomedicine. [ABSTRACT FROM AUTHOR]