G-quadruplex chemiluminescent sensor for the detection of specific nucleic acid sequences of pathogenic microorganisms
Gleb Bobkov 1, Pavel V. Filatov 1,2, Daria A. Gorbenko 1,2, Daler D. Dadazhanov 1, Tigran A. Vartanyan1,
Abstract
Efficient point-of-care (POC) molecular diagnostic techniques have the potential to revolutionize healthcare by providing a rapid and accurate diagnosis of infec- tious diseases. In this study, we propose the design of a peroxidase-like DNA-nanomachines (PxDm) that can selectively hybridize with specific analytes and produce readable outputs. We have chosen S.aureus, a pathogen responsible for a number of nosocomial infections, as the target analyte. For detection, we se- lected a specific gene that is constitutively expressed. The PxDm was equipped with three long analyte binding arms to tightly bind and unwind analytes. Only when all arms bind the analyte a G-quadruplex (G-4) structure is assembled. This structure can form a complex with hemin, which exhibits peroxidase activity. In the presence of the G-4/hemin complex, the chemiluminescence (CL) of luminol molecules, activated by hydrogen peroxide, is enhanced. We evaluated the CL kinetics for several minutes, comparing them with the background signal using a photon counting head. The results indicate that S.aureus was recognized with high selectivity and sensitivity up to zeptoM concentrations in a quartz cuvette. In the future, this system may become a full-fledged lab-on-a-chip for the detec- tion of marker-sequenced nucleic acids. The PxDm has several advantages over traditional diagnostic techniques, including its ability to detect analytes at ambient temperatures and its high selectivity and sensitivity.
The authors are grateful to ITMO University. We also thank the Ministry of educa- tion and science of the Russian federation No FSER-2022-0009 and Priority 2030 program.
File with abstract
Speaker
Gleb Bobkov
1Laboratory of Solution Chemistry of Advanced Materials and Technologies, ITMO university, Saint Petersburg, 2 SCAMT Institute, ITMO University, 49 Kronverksky pr., St. Petersburg 197101, Russian Federation
Russia
Discussion
Ask question
