Label-free Electrical Monitoring of Nucleic Acid Amplification with Integrated Hydrogel Ionic Diodes

Overview

Journal Mater Today Bio

Specialty Biomedical Engineering

Date 2022 May 24

PMID 35607416

Authors

Chenwei Xiong

Jie Li

Luyao Li

Long Chen

Rong Zhang

Xianqiang Mi

Yifan Liu

Affiliations

Soon will be listed here.

Abstract

We demonstrate here for the first time the utility of a monolithically integrated hydrogel ionic diode for label-free quantitative DNA detection and real-time monitoring of nucleic acid amplification. The hydrogel ionic diode presented herein, unlike nanomaterial-based field-effect biosensors, features high cost-effectiveness and convenient fabrication. This is realized by patterning a micrometer-sized heterojunction consisting of adjacent segments of polycationic and polyanionic hydrogels on a microfluidic chip through simple photocuring steps. The integrated diode rectifies ionic currents being sensitive to the charge of DNA adsorbed onto the polycationic chains through electrostatic associations. Based on the mechanism, we show that the ionic biosensor can electrically quantify DNA in a dynamic range relevant to typical nucleic acid amplification assays. Utilizing the device, we demonstrate the evaluation of a PCR assay amplifying a 500-bp DNA fragment of , an infection-causing pathogen, and real-time monitoring of an isothermal assay amplifying whole genome. We anticipate that the device could potentially pave the way for miniaturized optics-free platforms for quantifying nucleic acid amplification at point-of-care.

Citing Articles

Advanced materials for disease diagnostics.

Huang L, Qian K Mater Today Bio. 2023; 20:100676.

PMID: 37304576 PMC: 10250909. DOI: 10.1016/j.mtbio.2023.100676.

Recent Progress in Biomedical Sensors Based on Conducting Polymer Hydrogels.

Gamboa J, Paulo-Mirasol S, Estrany F, Torras J ACS Appl Bio Mater. 2023; 6(5):1720-1741.

PMID: 37115912 PMC: 10189742. DOI: 10.1021/acsabm.3c00139.

References

Li S, Jiang W, Huang J, Liu Y, Ren L, Zhuang L . Highly sensitive and specific diagnosis of COVID-19 by reverse transcription multiple cross-displacement amplification-labelled nanoparticles biosensor. Eur Respir J. 2020; 56(6). PMC: 7453731. DOI: 10.1183/13993003.02060-2020. View

Peker N, Couto N, Sinha B, Rossen J . Diagnosis of bloodstream infections from positive blood cultures and directly from blood samples: recent developments in molecular approaches. Clin Microbiol Infect. 2018; 24(9):944-955. DOI: 10.1016/j.cmi.2018.05.007. View

Tahamtan A, Ardebili A . Real-time RT-PCR in COVID-19 detection: issues affecting the results. Expert Rev Mol Diagn. 2020; 20(5):453-454. PMC: 7189409. DOI: 10.1080/14737159.2020.1757437. View

Cai B, Wang S, Huang L, Ning Y, Zhang Z, Zhang G . Ultrasensitive label-free detection of PNA-DNA hybridization by reduced graphene oxide field-effect transistor biosensor. ACS Nano. 2014; 8(3):2632-8. DOI: 10.1021/nn4063424. View

Han J, Kim K, Kim H, Chung T . Ionic circuits based on polyelectrolyte diodes on a microchip. Angew Chem Int Ed Engl. 2009; 48(21):3830-3. DOI: 10.1002/anie.200900045. View

Xiong C, Zhang B, Zhang R, Liu Y . An Experimental and Numerical Study of Polyelectrolyte Hydrogel Ionic Diodes: Towards Electrical Detection of Charged Biomolecules. Sensors (Basel). 2021; 21(24). PMC: 8707621. DOI: 10.3390/s21248279. View

He S, Tian J, Li X, Zhou Y, Xiao M, Zhang Y . Positive RT-PCR Test Results in 420 Patients Recovered From COVID-19 in Wuhan: An Observational Study. Front Pharmacol. 2020; 11:549117. PMC: 7577046. DOI: 10.3389/fphar.2020.549117. View

Liu Y, Yobas L . Slowing DNA Translocation in a Nanofluidic Field-Effect Transistor. ACS Nano. 2016; 10(4):3985-94. DOI: 10.1021/acsnano.6b00610. View

Dean F, Hosono S, Fang L, Wu X, Faruqi A, Bray-Ward P . Comprehensive human genome amplification using multiple displacement amplification. Proc Natl Acad Sci U S A. 2002; 99(8):5261-6. PMC: 122757. DOI: 10.1073/pnas.082089499. View

10.

Sarkar D, Liu W, Xie X, Anselmo A, Mitragotri S, Banerjee K . MoS₂ field-effect transistor for next-generation label-free biosensors. ACS Nano. 2014; 8(4):3992-4003. DOI: 10.1021/nn5009148. View

11.

Shen M, Li B, Li Y . Silicon nanowire field-effect-transistor based biosensors: from sensitive to ultra-sensitive. Biosens Bioelectron. 2014; 60:101-11. DOI: 10.1016/j.bios.2014.03.057. View

12.

Gudnason H, Dufva M, Bang D, Wolff A . Comparison of multiple DNA dyes for real-time PCR: effects of dye concentration and sequence composition on DNA amplification and melting temperature. Nucleic Acids Res. 2007; 35(19):e127. PMC: 2095797. DOI: 10.1093/nar/gkm671. View

13.

Ali M, Yameen B, Cervera J, Ramirez P, Neumann R, Ensinger W . Layer-by-layer assembly of polyelectrolytes into ionic current rectifying solid-state nanopores: insights from theory and experiment. J Am Chem Soc. 2010; 132(24):8338-48. DOI: 10.1021/ja101014y. View

14.

Ackermann M, Verleden S, Kuehnel M, Haverich A, Welte T, Laenger F . Pulmonary Vascular Endothelialitis, Thrombosis, and Angiogenesis in Covid-19. N Engl J Med. 2020; 383(2):120-128. PMC: 7412750. DOI: 10.1056/NEJMoa2015432. View

15.

Kellner M, Koob J, Gootenberg J, Abudayyeh O, Zhang F . SHERLOCK: nucleic acid detection with CRISPR nucleases. Nat Protoc. 2019; 14(10):2986-3012. PMC: 6956564. DOI: 10.1038/s41596-019-0210-2. View

16.

Sinha M, Jupe J, Mack H, Coleman T, Lawrence S, Fraley S . Emerging Technologies for Molecular Diagnosis of Sepsis. Clin Microbiol Rev. 2018; 31(2). PMC: 5967692. DOI: 10.1128/CMR.00089-17. View

17.

Chun H, Chung T . Iontronics. Annu Rev Anal Chem (Palo Alto Calif). 2015; 8:441-62. DOI: 10.1146/annurev-anchem-071114-040202. View

18.

Green N, Norton M . Interactions of DNA with graphene and sensing applications of graphene field-effect transistor devices: a review. Anal Chim Acta. 2014; 853:127-142. DOI: 10.1016/j.aca.2014.10.023. View

19.

Stine R, Robinson J, Sheehan P, Tamanaha C . Real-time DNA detection using reduced graphene oxide field effect transistors. Adv Mater. 2010; 22(46):5297-300. DOI: 10.1002/adma.201002121. View

20.

Liu Y, Yobas L . Label-free electrical quantification of amplified nucleic acids through nanofluidic diodes. Biosens Bioelectron. 2013; 50:78-83. DOI: 10.1016/j.bios.2013.06.013. View