Preparation and Application of Manufactured Electrochemical Sensors for the Multiplexed Detection

Abstract

Monkeypox is a zoonotic viral infection caused by MKPV (1). This disease causes skin lesions, fever and headache among other symptoms, which appear within a few days after the infection, and can led to the hospitalization of the infected person (2). Herein, we present the first 3D-printed electrochemical bio-device for multiplex analysis, enabling portable detection of the Monkeypox Virus (MKPV). The electrochemical device consists of two working biosensors, an immunosensor, and a genosensor, specifically designed for the detection of the protein A29 (antigen) and the DNA target of MKPV. The electrodes were manufactured using lab-made ultra-flexible conductive filaments composed of carbon black, recycled PLA from coffee capsules, and castor oil as plasticizer. The sensors created through 3D printing technology exhibited good reproducibility and repeatability of analytical responses. Furthermore, both the immunosensor and genosensor demonstrated excellent MKPV detection capabilities, with a linear range from 0.01 to 1.0 µmol L-1 for the antigen and 0.1 to 20.0 µmol L-1 for the DNA target. The biosensors achieved limits of detection of 2.7 nmol L-1 and 29.0 nmol L-1 for the immunosensor and genosensor, respectively. Interference tests conducted with the biosensors demonstrated their selectivity for MKPV. Moreover, analyses of fortified human serum samples showed recoveries close to 100%, confirming the absence of significant matrix effects in MKPV analysis. Therefore, the 3D-printed multiplex device represents a viable and highly promising alternative for on-site, portable, and rapid point-of-care MKPV monitoring.(1) Shafaati, M., Zandi, M., 2022. State-of-the-art on monkeypox virus: an emerging zoonotic disease. Infection 50, 1425–1430.(2) Halvaei, P., Zandi, S., Zandi, M., 2023. Biosensor as a novel alternative approach for early diagnosis of monkeypox virus. Int. J. Surg. 109, 50–52.