A novel approach for DNA extraction of white spot syndrome virus detection in penaeid shrimp

一种用于对虾白斑综合征病毒检测的DNA提取新方法

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Abstract

Penaeid shrimp and other aquatic crustaceans are highly susceptible to infection by the White Spot Syndrome Virus (WSSV). In recent years, genomic DNA extraction techniques have become central to many molecular diagnostics and technologies. DNA extraction kits were not included in the comparison because they often lack the flexibility and cost-effectiveness needed for large-scale applications in aquaculture research. This study introduces a novel Dimethyl Sulfoxide (DMSO)-based DNA extraction method that is simple, rapid, and equally sensitive, marking a unique advancement in aquaculture research. We used identical sample volumes to compare the DMSO-based method with the traditional phenol-chloroform and Guanidium Hydrochloride techniques. In the study, the DNA yield obtained using DMSO was 378.4 ng/µL, which was significantly higher compared to the other two extraction methods. The phenol-chloroform method yielded 267.8 ng/µL, while the GHCL method produced 151.2 ng/µL. The DMSO method has a lower detection limit and can detect DNA at concentrations as low as 10⁻⁹, whereas the GHCL and Phenol-Chloroform methods have detection limits of 10⁻⁷. These results suggest that DMSO provides a higher DNA yield and has a lower detection limit than the phenol-chloroform and GHCL methods, indicating its potential for more efficient DNA extraction. Therefore, our study introduces a high-quality genomic DNA extraction protocol applicable to the diagnosis of diseases in other marine organisms.

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