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Published on: October 6, 2017
Chemiluminescent microplate-based assay of DNA based on isothermal circular strand-displacement polymerization
Anton M Solovjev1, Sergei A Kurzeev1, Ivan Yu Sakharov1
1Department of Chemistry, Lomonosov Moscow State University, Leninskie Gory, Bldg.1, Moscow, 119991, Russia.
Abstract:
Sensitive microplate-based chemiluminescent assay coupled with isothermal circular strand-displacement polymerization reaction (ICSDPR) for DNA detection was developed. The assay sensitivity was improved using a triple amplification strategy based on employment of ICSDPR, streptavidin-polyperoxidase conjugate and an enhanced chemiluminescent reaction. To increase the accuracy all stages of the assay (one-pot format) were carried out in the same microplate well. The proposed assay detected the target DNA in fM-pM ranges and distinguish target DNA from related mismatched DNAs, demonstrating high sensitivity and high selectivity. This format is highly suitable for the development of sensitive and high-throughput kits of nucleic acids which can be easily automated.
