Development of ultrasensitive genosensor targeting pathogenic Leptospira DNA detection in artificial urine
Issued Date
2024-12-10
Resource Type
ISSN
00134686
Scopus ID
2-s2.0-85206094825
Journal Title
Electrochimica Acta
Volume
507
Rights Holder(s)
SCOPUS
Bibliographic Citation
Electrochimica Acta Vol.507 (2024)
Suggested Citation
Suwannin P., Jangpatarapongsa K., Frías I.A.M., Polpanich D., Techakasikornpanich M., Elaissari A., Errachid A. Development of ultrasensitive genosensor targeting pathogenic Leptospira DNA detection in artificial urine. Electrochimica Acta Vol.507 (2024). doi:10.1016/j.electacta.2024.145145 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/101668
Title
Development of ultrasensitive genosensor targeting pathogenic Leptospira DNA detection in artificial urine
Author's Affiliation
Corresponding Author(s)
Other Contributor(s)
Abstract
In the last decades, to access the stages of leptospirosis pathogenicity, site of infection, progression, monitor disease, and after-treatment were required especially non-invasive techniques. This becomes one of the significant goals for researchers. In this study, we developed a highly sensitive DNA-based electrochemical sensor to detect pathogenic leptospires in urine samples using a gold screen-printed electrode. The gold working electrode was electrodeposited with diazonium salt (CMA) with external carboxylic acid from the surface, and the specific Loa22 single-stranded DNA (ssDNA) probe to Leptospira interrogans was immobilized through a carbodiimide chemical reaction. The modified working electrode was characterized by contact angle measurement, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). After that, targeted DNA testing showed the EIS change in the range of 5 ag mL-1 - 16 fg mL-1 with a correlation R2 of 0.9638. The high sensitivity electrochemical sensor indicated a lower limit of detection to the attomolar level specifically 5 ag mL-1. Additionally, the developed genosensor exhibited high specificity without crossing other bacteria present in urine such as Escherichia coli, Staphylococcus aureus, and S. typhi as well as non-pathogenic leptospires L. biflexa serovar Patoc. Leptospires DNA was analyzed in both buffer solution and diluted artificial urine and detection in diluted artificial urine 1:1,000 was possible despite salt interferences. This study provides a highly sensitive lower detection platform for leptospirosis, offering potential for further development as a portable and point-of-care testing tool.