Electrochemical aptasensor detection of electron transfer flavoprotein subunit beta for leptospirosis diagnosis
3
Issued Date
2023-01-01
Resource Type
ISSN
00032654
eISSN
13645528
Scopus ID
2-s2.0-85169505991
Journal Title
Analyst
Rights Holder(s)
SCOPUS
Bibliographic Citation
Analyst (2023)
Suggested Citation
Kositanont U., Srisawat C., Sripinitchai S., Thawornkuno C., Chaibun T., Karunaithas S., Promptmas C., Lertanantawong B. Electrochemical aptasensor detection of electron transfer flavoprotein subunit beta for leptospirosis diagnosis. Analyst (2023). doi:10.1039/d3an01064c Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/89590
Title
Electrochemical aptasensor detection of electron transfer flavoprotein subunit beta for leptospirosis diagnosis
Other Contributor(s)
Abstract
Electron transfer flavoprotein subunit beta (ETFB) of Leptospira interrogans is a biomarker for diagnosing leptospiral infection. Thus, the ETFB-specific nuclease-resistant RNA aptamer ETFB3-63 was developed and used in an electrochemical aptasensor to assay ETFB. Although the majority of reported biosensors detect various genes and antibodies of L. interrogans, this is the first attempt to construct an electrochemical biosensor to detect ETFB protein for the diagnosis of leptospiral infection. The ETFB protein can be detected without any extraction phase. In this assay, a single-stranded DNA probe complementary to the ETFB3-63 sequence was immobilized on a screen-printed carbon electrode (SPCE). The aptamer was then incubated and hybridized with the antisense probe on the SPCE. In the presence of ETFB, the aptamer dissociates from the aptamer/probe complex on the SPCE to bind with the protein. Methylene blue was then added to intercalate with the remaining hybridized aptamers, and its signal was measured using differential pulse voltammetry. The signal arising from the intercalated methylene blue decreased with increasing concentration of ETFB, showing a linear response in the range of 50-500 nM of ETFB and 10 to 109 leptospira cells per mL, respectively. The aptasensor signal was also specific to L. interrogans but not to 12 related bacteria tested. In addition, the aptasensor showed similar performance in detecting ETFB spiked in human serum to that in buffer, indicating that proteins in the serum do not interfere with the assay. Therefore, this assay has great potential to develop into a point-of-care electrochemical device that is accurate, cost-effective, and user-friendly for leptospirosis diagnosis.