A DNA Barcode‐Based Aptasensor Enables Rapid Testing of Porcine Epidemic Diarrhea Viruses in Swine Saliva Using Electrochemical Readout

Abstract

Pen-side testing of farm animals for infectious diseases is critical for preventing transmission in herds and providing timely intervention. However, most existing pathogen tests have to be conducted in centralized labs with sample-to-result times of 2-4 days. Herein we introduce a test that uses a dual-electrode electrochemical chip (DEE-Chip) and a barcode-releasing electroactive aptamer for rapid on-farm detection of porcine epidemic diarrhea viruses (PEDv). The sensor exploits inter-electrode spacing reduction and active field mediated transport to accelerate barcode movement from electroactive aptamers to the detection electrode, thus expediting assay operation. The test yielded a clinically relevant limit-of-detection of 6 nM (0.37 μg mL À 1 ) in saliva-spiked PEDv samples. Clinical evaluation of this biosensor with 12 porcine saliva samples demonstrated a diagnostic sensitivity of 83 % and specificity of 100 % with a concordance value of 92 % at an analysis time of one hour.

Development of Duplex LAMP Technique for Detection of Porcine Epidemic Diarrhea Virus (PEDV) and Porcine Circovirus Type 2 (PCV 2)

“…In this study, a prototype for a duplex LAMP-LFD assay relevant to Technology Readiness Level 3 (TRL 3) was developed for simultaneous detection of PEDV and PCV 2 in the same specimens. According to the LOD, the duplex LAMP-LFD was 10-times more sensitive than PCR-based AGE and LAMP-based AGE, which correlates with previous reports [ 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 ]. The high analytical specificity of the duplex LAMP-LFD resulted from primers and DNA probes that were uniquely designed from the conserved regions on the spike and ORF genes for PEDV and PCV2, respectively.…”

Aptamer‐functionalized field‐effect transistor biosensors for disease diagnosis and environmental monitoring

“…[61][62][63][64] Based on these prominent characteristics, aptamers could provide ultrahigh sensitivity and specificity, and they have become a powerful tool for advanced biological and biomedical applications. [65][66][67][68][69]…”

Os(ii/iii) complex supports pH-insensitive electrochemical DNA-based sensing with superior operational stability than the benchmark methylene blue reporter

“…Electrochemical, DNA-based (E-DNA) sensors rely on binding/hybridization-induced conformational changes to generate an electrical signal, enabling single-point 1 or continuous 2 molecular measurements. Electrochemical, aptamer-based (E-AB) sensors represent a specific type of E-DNA sensors that enable the continuous and real-time monitoring of molecular targets.…”