Chemiluminescent microplate-based assay of DNA based on isothermal circular strand-displacement polymerization reaction (ICSDPR)

Anton M Solovjev; Sergei A Kurzeev; Ivan Yu Sakharov

doi:10.1016/j.talanta.2020.120895

Chemiluminescent microplate-based assay of DNA based on isothermal circular strand-displacement polymerization reaction (ICSDPR)

Talanta. 2020 Aug 1:215:120895. doi: 10.1016/j.talanta.2020.120895. Epub 2020 Mar 6.

Authors

Anton M Solovjev¹, Sergei A Kurzeev¹, Ivan Yu Sakharov²

Affiliations

¹ Department of Chemistry, Lomonosov Moscow State University, Leninskie Gory, Bldg.1, Moscow, 119991, Russia.
² Department of Chemistry, Lomonosov Moscow State University, Leninskie Gory, Bldg.1, Moscow, 119991, Russia. Electronic address: sakharovivan@gmail.com.

PMID: 32312440
DOI: 10.1016/j.talanta.2020.120895

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.

Keywords: Circular strand-displacement polymerization reaction; DNA oligonucleotide; Enhanced chemiluminescence; Isothermal amplification; Microplate-based assay; Polyperoxidase conjugate.

MeSH terms

Biosensing Techniques*
DNA, Viral / analysis*
Luminescent Measurements*
Nucleic Acid Amplification Techniques*

Substances

DNA, Viral