Toward the early diagnosis of tuberculosis: A gold particle-decorated graphene-modified paper-based electrochemical biosensor for Hsp16.3 detection

• Published in: Talanta (Oxford) Vol. 267; p. 125210
• Main Authors: Pornprom, Thimpika, Phusi, Naruedon, Thongdee, Paptawan, Pakamwong, Bongkochawan, Sangswan, Jidapa, Kamsri, Pharit, Punkvang, Auradee, Suttisintong, Khomson, Leanpolchareanchai, Jiraporn, Hongmanee, Poonpilas, Lumjiaktase, Putthapoom, Jampasa, Sakda, Chailapakul, Orawon, Pungpo, Pornpan
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 15-01-2024
• Subjects: Early diagnosis; Half-sandwich format; Heat shock protein 16.3; Paper-based electrochemical biosensor; Tuberculosis; Half-sandwich format; Early diagnosis; Tuberculosis; Heat shock protein 16.3; Paper-based electrochemical biosensor
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2023.125210

Tuberculosis (TB) currently remains a major life-threatening disease as it can be fatal if not treated properly or in a timely manner. Herein, we first describe a disposable and cost-effective paper-based electrochemical biosensor based on a gold particle-decorated carboxyl graphene (AuPs/GCOOH)-modified electrode for detecting heat shock protein (Hsp16.3), which is a specific biomarker indicating the onset of TB infection. The device pattern was first engineered to facilitate detection procedures and printed on low-cost filter paper to create hydrophobic and hydrophilic regions using a wax printing technique. Immunoassays proceeded in a half-sandwich format because it is a reagent-less approach and requires no labeling step. The fabrication of the immunosensor began with GCOOH drop casting, the electrochemical deposition of AuPs, and the establishment of a biorecognition layer against Hsp16.3 utilizing 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS)-sulfo standard chemistry. The appearance of Hsp16.3 resulted in a substantial decrease in the electrochemical signal response of the redox probe employed [Fe (CN)6]3−/4− due to the created immunocomplexes that possess insulation properties. GCOOH enables direct antibody immobilization, and AuPs enhance the electrochemical properties of the sensor. This proposed immunosensor, while requiring only a miniscule sample volume (5 μL), achieved superior performance in terms of the limit of detection, measuring at 0.01 ng/mL. Our platform was confirmed to be highly specific to Hsp16.3 and can rapidly detect TB-infected sera without necessitating any pre-enrichment (20 min), making it an alternative and particularly suitable for the early diagnosis of TB in resource-scarce countries.
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•An engineered paper-based electrochemical biosensor was first proposed for detection of Hsp 16.3 to diagnose TB infection early.•AuPs and GCOOH decorated on the working area enhanced the sensing capabilities of the established PAD.•Laten and active TB-infected human sera can be directly detected without requiring any pre-enrichment.•The established biosensor was sensitive, low-cost, portable, and disposable.