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A CRISPR-Cas13a powered electrochemical sensor based on reduced graphene oxide, polypyrrole and gold nanoparticles nanocomposites for PEDV detection

Zhaofan Zhou,Ye Li,Dongya Wen,Xuyu Zhang,Ni Li,Jiangbing Shuai,Xiaofeng Zhang,Xiaoye Zheng,Zhaojun Ban,Farouk Amr,Xiaoping Yu,Jun Huang

Porcine epidemic diarrhea virus (PEDV) is a highly infectious pathogen responsible for porcine epidemic diarrhea, which causes severe diarrhea and high mortality rates in neonatal piglets, leading to substantial economic losses in the swine industry. Therefore, there is an urgent need for rapid, sensitive, and accurate detection methods for PEDV. In this study, we develop a rapid, ultrasensitive electrochemical CRISPR-based biosensor for detecting PEDV with high sensitivity and specificity. The sensor integrates reduced graphene oxide-polypyrrole‑gold nanoparticle (rGO-PPy-AuNP) nanocomposites to enhance sensitivity and CRISPR-Cas13a for target-specific recognition. This dual-signal amplification strategy achieves an excellent analytical detection limit of 1.01 fg/mL and high linearity (R 2  = 0.9979) across a broad dynamic range (0.005–100,000 pg/mL) within 45 min, eliminating the need for nucleic acid amplification. By synergizing Cas13a-assisted signal enhancement with nanoparticle-mediated electron transfer, the biosensor outperforms conventional methods in sensitivity while maintaining excellent stability, and specificity, demonstrating strong potential for PEDV detection in clinical diagnostics.