Field-usable aptamer‑gold nanoparticles-based colorimetric sensor for rapid detection of white spot syndrome virus in shrimp

Abstract White spot syndrome virus (WSSV) is one of the most threatening viral pathogens of shrimp worldwide. VP28, the major capsid protein of WSSV, is reported to play an essential role in the interaction with the host cells. Usually, the diagnostic test of WSSV is performed by one-step PCR, which is neither field-usable nor rapid enough. Therefore, the development of early diagnostic tests is imperative for the management of disease and to prevent huge economic losses. In this work, a rapid colorimetric aptasensor with high selectivity and sensitivity was introduced. First, a new ssDNA aptamer for the detection of WSSV was designed from a random pool of aptamer sequences based on the docking score and bioconjugate free energy, and then, evaluated experimentally. The designed aptamer was used to prepare an aptasensor conjugate gold nanoparticles (AuNPs) as a recognition element that increased the resistance of AuNPs to salt-induced aggregation. Therefore, the AuNPs with spherical morphology and average particle size of about 20 nm were synthesized for this reason. The results showed that the aptamer didn't attach to the other pathogens (i.e., IHHN and Vibrio harveyi) and healthy shrimp cells, and remained stable. Interestingly, the gray-blue color of aptasensor was seen only by the presence of WSSV, indicating the aptasensor assay showed good specificity to WSSV. The limit of detection (LOD) of the aptasensor was 104 copies of WSSV that could be detected only utilizing naked eyes. It was concluded that the designed aptamer had excellent potential for the detection of WSSV as a rapid visual colorimetric assay in aquaculture shrimp farmers.