{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T21:04:02Z","timestamp":1780607042100,"version":"3.54.1"},"reference-count":45,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,4,18]],"date-time":"2023-04-18T00:00:00Z","timestamp":1681776000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Herein, we report results of the studies relating to the development of an impedimetric, magnetic bead-assisted supersandwich DNA hybridization assay for ultrasensitive detection of Neisseria gonorrhoeae, the causative agent of a sexually transmitted infection (STI), gonorrhea. First, a conductive ink was formulated by homogenously dispersing carboxylated multiwalled carbon nanotubes (cMWCNTs) in a stable emulsion of terpineol and an aqueous suspension of carboxymethyl cellulose (CMC). The ink, labeled C5, was coated onto paper substrates to fabricate C5@paper conductive electrodes. Thereafter, a magnetic bead (MB)-assisted supersandwich DNA hybridization assay was optimized against the porA pseudogene of N. gonorrhoeae. For this purpose, a pair of specific 5\u2032 aminated capture probes (SCP) and supersandwich detector probes (SDP) was designed, which allowed the enrichment of target gonorrheal DNA sequence from a milieu of substances. The SD probe was designed such that instead of 1:1 binding, it allowed the binding of more than one T strand, leading to a \u2018ladder-like\u2019 DNA supersandwich structure. The MB-assisted supersandwich assay was integrated into the C5@paper electrodes for electrochemical analysis. The C5@paper electrodes were found to be highly conductive by a four-probe conductivity method (maximum conductivity of 10.1 S\u00b7cm\u22121). Further, the biosensing assay displayed a wide linear range of 100 aM-100 nM (109 orders of magnitude) with an excellent sensitivity of 22.6 k\u03a9\u00b7(log[concentration])\u22121. The clinical applicability of the biosensing assay was assessed by detecting genomic DNA extracted from N. gonorrhoeae in the presence of DNA from different non-gonorrheal bacterial species. In conclusion, this study demonstrates a highly sensitive, cost-effective, and label-free paper-based device for STI diagnostics. The ink formulation prepared for the study was found to be highly thixotropic, which indicates that the paper electrodes can be screen-printed in a reproducible and scalable manner.<\/jats:p>","DOI":"10.3390\/bios13040486","type":"journal-article","created":{"date-parts":[[2023,4,18]],"date-time":"2023-04-18T05:18:13Z","timestamp":1681795093000},"page":"486","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Conductive Ink-Coated Paper-Based Supersandwich DNA Biosensor for Ultrasensitive Detection of Neisseria gonorrhoeae"],"prefix":"10.3390","volume":"13","author":[{"given":"Niharika","family":"Gupta","sequence":"first","affiliation":[{"name":"Department of Biotechnology, Delhi Technological University, Delhi 110042, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D.","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Applied Chemistry, Delhi Technological University, Delhi 110042, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asmita","family":"Das","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Delhi Technological University, Delhi 110042, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seema","family":"Sood","sequence":"additional","affiliation":[{"name":"Department of Microbiology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110016, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0957-2684","authenticated-orcid":false,"given":"Bansi D.","family":"Malhotra","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Delhi Technological University, Delhi 110042, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,18]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (WHO) (2023, February 24). 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