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PCR-assisted impedimetric biosensor for colibactin-encoding pks genomic island detection in E. coli samples.
| Content Provider | Europe PMC |
|---|---|
| Author | Solis-Marcano, Nadja E. Morales-Cruz, Myreisa Vega-Hernández, Gabriela Gómez-Moreno, Ramón Binder, Claudia Baerga-Ortiz, Abel Priest, Craig Cabrera, Carlos R. |
| Abstract | A fast PCR-assisted impedimetric biosensor was developed for the selective detection of the clbN gene from the polyketide synthase (pks) genomic island in real Escherichia coli samples. This genomic island is responsible for the production of colibactin, a harmful genotoxin that has been associated with colorectal cancer. The experimental protocol consisted of immobilizing the designated forward primer onto an Au electrode surface to create the sensing probe, followed by PCR temperature cycling in blank, positive, and negative DNA controls. Target DNA identification was possible by monitoring changes in the system’s charge transfer resistance values (Rct) before and after PCR treatment through electrochemical impedance spectroscopy (EIS) analysis. Custom-made, flexible gold electrodes were fabricated using chemical etching optical lithography. A PCR cycle study determined the optimum conditions to be at 6 cycles providing fast results while maintaining a good sensitivity. EIS data for the DNA recognition process demonstrated the successful distinction between target interaction resulting in an increase in resistance to charge transfer (Rct) percentage change of 176% for the positive DNA control vs. 21% and 20% for the negative and non-DNA-containing controls, respectively. Results showed effective fabrication of a fast, PCR-based electrochemical biosensor for the detection of pks genomic island with a calculated limit of detection of 17 ng/μL.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00216-021-03404-6. |
| ISSN | 16182642 |
| Journal | Analytical and Bioanalytical Chemistry |
| Volume Number | 413 |
| PubMed Central reference number | PMC8159250 |
| Issue Number | 18 |
| PubMed reference number | 34046698 |
| e-ISSN | 16182650 |
| DOI | 10.1007/s00216-021-03404-6 |
| Language | English |
| Publisher | Springer Berlin Heidelberg |
| Publisher Date | 2021-05-27 |
| Publisher Place | Berlin/Heidelberg |
| Access Restriction | Open |
| Rights License | This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. © Springer-Verlag GmbH Germany, part of Springer Nature 2021 |
| Subject Keyword | Electrochemical impedance spectroscopy DNA biosensor Polyketide synthase genomic island Polymerase chain reaction (PCR) |
| Content Type | Text |
| Resource Type | Article |
| Subject | Analytical Chemistry Biochemistry |
