{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T02:59:37Z","timestamp":1789441177807,"version":"build-2803163510"},"reference-count":41,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2020,7,3]],"date-time":"2020-07-03T00:00:00Z","timestamp":1593734400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100003896","name":"Henry M. Jackson Foundation","doi-asserted-by":"publisher","award":["Contract No. 895748"],"award-info":[{"award-number":["Contract No. 895748"]}],"id":[{"id":"10.13039\/100003896","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007680","name":"Naval Medical Research Center","doi-asserted-by":"publisher","award":["Contract No.: N3239818P0305"],"award-info":[{"award-number":["Contract No.: N3239818P0305"]}],"id":[{"id":"10.13039\/100007680","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology of Taiwan","doi-asserted-by":"publisher","award":["MOST107-2313-B-005-006-MY3"],"award-info":[{"award-number":["MOST107-2313-B-005-006-MY3"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministry of Education in Taiwan","award":["Higher Education Sprout Project"],"award-info":[{"award-number":["Higher Education Sprout Project"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Developing rapid and sensitive diagnostic methods for dengue virus (DENV) infection is of prime priority because DENV infection is the most prevalent mosquito-borne viral disease. This work proposes an electrochemical impedance spectroscopy (EIS)-based genosensor for the label-free and nucleic acid amplification-free detection of extracted DENV RNA intended for a sensitive diagnosis of DENV infection. A concentration ratio of 0.04 mM 6-mercaptohexanoic acid (MHA) to 1 mM 6-mercapto-1-hexanol (MCH) was selected to modify thin-film gold electrodes as a link to control the coverage of self-designed probe DNA (pDNA) at a density of 4.5 \u00b1 0.4 \u00d7 1011 pDNA\/cm2. The pDNA\/MHA\/MCH-modified genosensors are proven to improve the hybridization efficiency of a synthetic 160-mer target DNA (160mtDNA) with a 140-mer electrode side overhang as compared to other MHA\/MCH ratio-modified genosensors. The MHA(0.04 mM)\/MCH(1 mM)-modified genosensors also present good hybridization efficiency with the extracted DENV serotype 1 (DENV1) RNA samples, having the same electrode side overhangs with the 160mtDNA, showing a low detection limit of 20 plaque forming units (PFU)\/mL, a linear range of 102\u2013105 PFU\/mL and good selectivity for DENV1. The pDNA density-controlled method has great promise to construct sensitive genosensors based on the hybridization of extracted DENV nucleic acids.<\/jats:p>","DOI":"10.3390\/s20133728","type":"journal-article","created":{"date-parts":[[2020,7,6]],"date-time":"2020-07-06T09:49:11Z","timestamp":1594028951000},"page":"3728","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["A Label-Free Impedimetric Genosensor for the Nucleic Acid Amplification-Free Detection of Extracted RNA of Dengue Virus"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8683-3796","authenticated-orcid":false,"given":"Ching-Chou","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Bio-Industrial Mechatronics Engineering, National Chung Hsing University, No. 145, Xingda Rd., South Dist., Taichung City 402, Taiwan"},{"name":"Innovation and Development Center of Sustainable Agriculture, National Chung Hsing University, No. 145, Xingda Rd., South Dist., Taichung City 402, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao-Yu","family":"Yen","sequence":"additional","affiliation":[{"name":"Department of Bio-Industrial Mechatronics Engineering, National Chung Hsing University, No. 145, Xingda Rd., South Dist., Taichung City 402, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu-Ting","family":"Lai","sequence":"additional","affiliation":[{"name":"Department of Bio-Industrial Mechatronics Engineering, National Chung Hsing University, No. 145, Xingda Rd., South Dist., Taichung City 402, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guey-Chuen","family":"Perng","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, No.1, University Rd., Tainan City 701, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng-Rei","family":"Lee","sequence":"additional","affiliation":[{"name":"Viral &amp; Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuenn-Jue","family":"Wu","sequence":"additional","affiliation":[{"name":"Viral &amp; Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, MD 20910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1016\/j.pt.2018.01.005","article-title":"Revamping Mosquito-borne Disease Control to Tackle Future Threats","volume":"34","author":"Fernandes","year":"2018","journal-title":"Trends Parasitol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"70","DOI":"10.3389\/fphys.2012.00070","article-title":"Eventual role of asymptomatic cases of dengue for the introduction and spread of dengue viruses in non-endemic regions","volume":"3","author":"Chastel","year":"2012","journal-title":"Front Physiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1111\/jtm.12052","article-title":"Dengue fever and international travel","volume":"20","author":"Ratnam","year":"2013","journal-title":"J. Travel Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1016\/j.tim.2013.03.003","article-title":"Urbanization and geographic expansion of zoonotic arboviral diseases: Mechanisms and potential strategies for prevention","volume":"21","author":"Weaver","year":"2013","journal-title":"Trends Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1136\/pgmj.2004.019638","article-title":"Dengue viral infections","volume":"80","author":"Malavige","year":"2004","journal-title":"Postgrad. Med. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"S30","DOI":"10.1038\/nrmicro2459","article-title":"Evaluation of diagnostic tests: Dengue","volume":"8","author":"Peeling","year":"2010","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.coviro.2016.05.009","article-title":"Novel concept on antiviral strategies to dengue","volume":"18","author":"Lo","year":"2016","journal-title":"Curr. Opin. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.bios.2017.04.011","article-title":"A bead-based immunofluorescence-assay on a microfluidic dielectrophoresis platform for rapid dengue virus detection","volume":"95","author":"Iswardy","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.aca.2018.04.005","article-title":"An electrochemical peptide sensor for detection of dengue fever biomarker NS1","volume":"1026","author":"Lim","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.aca.2018.04.032","article-title":"Development of a portable and disposable NS1 based electrochemical immunosensor for early diagnosis of dengue virus","volume":"1026","author":"Nawaz","year":"2018","journal-title":"Anal. Chim. Acta"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/j.talanta.2019.01.058","article-title":"A new immunoassay of hybrid nanomater conjugated to aptamers for the detection of dengue virus","volume":"197","author":"Basso","year":"2019","journal-title":"Talanta"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.aca.2012.03.017","article-title":"Development of an electrochemical membrane-based nanobiosensor for ultrasensitive detection of dengue virus","volume":"725","author":"Cheng","year":"2012","journal-title":"Anal. Chim. Acta"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.diagmicrobio.2017.08.019","article-title":"Evaluation of OneStep Dengue NS1 RapiDip InstaTest and OneStep Dengue Fever IgG\/IgM RapiCard InstaTest during the course of a dengue type 1 epidemic","volume":"89","author":"Vickers","year":"2017","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"7774","DOI":"10.3390\/s130607774","article-title":"Impedimetric DNA biosensor based on a nanoporous alumina membrane for the detection of the specific oligonucleotide sequence of dengue virus","volume":"13","author":"Deng","year":"2013","journal-title":"Sensors"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.electacta.2016.02.116","article-title":"Impedimetric Dengue Biosensor based on Functionalized Graphene Oxide Wrapped Silica Particles","volume":"194","author":"Jin","year":"2016","journal-title":"Electrochim. Acta"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2211","DOI":"10.1007\/s00604-017-2249-5","article-title":"A needle-like Cu2CdSnS4 alloy nanostructure-based integrated electrochemical biosensor for detecting the DNA of Dengue serotype 2","volume":"184","author":"Odeh","year":"2017","journal-title":"Microchim. Acta"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"15562","DOI":"10.3390\/s150715562","article-title":"A sensitive and selective label-Free electrochemical DNA biosensor for the detection of specific dengue virus serotype 3 sequences","volume":"15","author":"Oliveira","year":"2015","journal-title":"Sensors"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.bios.2017.05.047","article-title":"A genosensor for detection of consensus DNA sequence of Dengue virus using ZnO\/Pt-Pd nanocomposites","volume":"97","author":"Singhal","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.bios.2016.12.008","article-title":"Electrospun manganese (III) oxide nanofiber based electrochemical DNA-nanobiosensor for zeptomolar detection of dengue consensus primer","volume":"90","author":"Tripathy","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_21","unstructured":"Sharp, T.M., Perez-Padilla, J., and Waterman, S.H. (2019, October 26). Travel-Related Infectious Diseases, Available online: https:\/\/wwwnc.cdc.gov\/travel\/yellowbook\/2020\/travel-related-infectious-diseases\/dengue."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3939","DOI":"10.1128\/JVI.02484-07","article-title":"Role of dendritic cells in antibody-dependent enhancement of dengue virus infection","volume":"82","author":"Boonnak","year":"2008","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Balsitis, S.J., Williams, K.L., Lachica, R., Flores, D., Kyle, J.L., Mehlhop, E., Johnson, S., Diamond, M.S., Beatty, P.R., and Harris, E. (2010). Lethal antibody enhancement of dengue disease in mice is prevented by Fc modification. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000790"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3316","DOI":"10.1016\/j.snb.2017.09.159","article-title":"Point-of-care tests: A review of advances in the emerging diagnostic tools for dengue virus infection","volume":"255","author":"Darwish","year":"2018","journal-title":"Sens. Actuator B-Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"111511","DOI":"10.1016\/j.bios.2019.111511","article-title":"Electrochemical biosensing of mosquito-borne viral disease, dengue: A review","volume":"142","author":"Anusha","year":"2019","journal-title":"Biosens. Bioelectron."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"113463","DOI":"10.1016\/j.jelechem.2019.113463","article-title":"Electrochemical voltammetric behaviors of synthetic dengue virus RNAs at ITO sensing electrode","volume":"851","author":"Cui","year":"2019","journal-title":"J. Electroanal. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"13458","DOI":"10.1021\/acs.analchem.9b02455","article-title":"Selective determination of isothermally amplified Zika virus RNA using a universal DNA-hairpin probe in less than 1 h","volume":"91","author":"Lynch","year":"2019","journal-title":"Anal. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.bios.2018.02.059","article-title":"A universal and label-free impedimetric biosensing platform for discrimination of single nucleotide substitutions in long nucleic acid strands","volume":"100","author":"Mills","year":"2018","journal-title":"Biosens. Bioelectron."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.jelechem.2014.08.026","article-title":"Improving electrochemical biosensor performance by understanding the influence of target DNA length on assay sensitivity","volume":"732","author":"Corrigan","year":"2014","journal-title":"J. Electroanal. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7867","DOI":"10.1021\/ac501800q","article-title":"Impedimetric DNA detection-steps forward to sensorial application","volume":"86","author":"Riedel","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.aca.2005.08.046","article-title":"Impedance spectral studies of self-assembly of alkanethiols with different chain lengths using different immobilization strategies on Au electrodes","volume":"554","author":"Ding","year":"2005","journal-title":"Anal. Chim. Acta"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4670","DOI":"10.1021\/ac980037q","article-title":"Electrochemical Quantitation of DNA Immobilized on Gold","volume":"70","author":"Steel","year":"1998","journal-title":"Anal. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"8916","DOI":"10.1021\/ja9719586","article-title":"Characterization of DNA Probes Immobilized on Gold Surfaces","volume":"119","author":"Herne","year":"1997","journal-title":"J. Am. Chem. Soc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.snb.2014.11.078","article-title":"An ultrasensitive label-free electrochemical impedimetric DNA biosensing chip integrated with a DC-biased AC electroosmotic vortex","volume":"209","author":"Wu","year":"2015","journal-title":"Sens. Actuator B-Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3758","DOI":"10.1039\/b305867k","article-title":"Multiple voltammetric waves for reductive desorption of cysteine and 4-mercaptobenzoic acid monolayers self-assembled on gold substrates","volume":"5","author":"Arihara","year":"2003","journal-title":"Phys. Chem. Chem. Phys."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0166-0934(99)00166-4","article-title":"Quantitative detection of dengue 2 virus using fluorogenic RT-PCR based on 3%-noncoding sequence","volume":"86","author":"Houng","year":"2000","journal-title":"J. Virol. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1128\/JCM.00842-06","article-title":"Development of multiplex real-time reverse transcriptase PCR assays for Detecting Eight Medically Important Flaviviruses in Mosquitoes","volume":"45","author":"Chao","year":"2007","journal-title":"J. Clin. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.trac.2015.01.005","article-title":"An introduction to dengue-disease diagnostics","volume":"67","author":"Darwish","year":"2015","journal-title":"TrAC Trend. Anal. Chem."},{"key":"ref_39","first-page":"7520","article-title":"Microfluidic biosensor for the serotype-specific detection of dengue virus RNA","volume":"77","author":"Zaytseva","year":"2005","journal-title":"Anal. Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"215501","DOI":"10.1088\/0957-4484\/20\/21\/215501","article-title":"A method of layer-by-layer gold nanoparticle hybridization in a quartzcrystal microbalance DNA sensing system used to detect dengue virus","volume":"20","author":"Chen","year":"2009","journal-title":"Nanotechnology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"127854","DOI":"10.1016\/j.snb.2020.127854","article-title":"A non-optical multiplexed PCR diagnostic platform for serotype-specific detection of dengue virus","volume":"310","author":"Yin","year":"2020","journal-title":"Sens. Actuator B-Chem."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/13\/3728\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:47:02Z","timestamp":1760176022000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/13\/3728"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,3]]},"references-count":41,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2020,7]]}},"alternative-id":["s20133728"],"URL":"https:\/\/doi.org\/10.3390\/s20133728","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,3]]}}}