A paper-based electrochemical aptasensor functionalized with silver nanoparticles for the detection of African swine fever virus p30 protein.

Abstract

African swine fever (ASF) is a devastating viral disease of domestic pigs and wild boars. The etiological agent, African swine fever virus (ASFV), continues to pose a serious threat to the global swine industry, causing profound economic losses in many countries. Therefore, early and reliable detection of ASFV is essential to curb its transmission and mitigate the economic damage it causes. In the present study, the ASFV protein p30, which is expressed within 2-4 h post-infection and endures throughout the viral replication cycle, was expressed and purified. An aptamer targeting the ASFV p30 protein was screened and validated, which was then used to fabricate an electrochemical paper-based analytical device (PAD) aptasensor. To enhance the aptasensor's conductivity, silver nanoparticles were embedded, and the aptamer was immobilized on the working electrode for ASFV p30 detection. Optimization included testing various protein concentrations and characterizing the resulting electrochemical perturbations using electrochemical impedance spectroscopy (EIS). The optimized biosensor enabled sensitive detection of ASFV p30 at low concentrations, achieving a limit of detection (LOD) of 1.09 ng/mL. These findings demonstrate the high sensitivity of the developed aptasensor and highlight its potential as a rapid and reliable platform for ASFV p30 protein detection. Overall, the proposed aptasensor provides a promising technical foundation for the development of rapid, sensitive, and accessible diagnostic approaches for ASFV.

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