EvaGreen-based real-time PCR assay for sensitive detection of salmonid alphavirus

In this study, we developed a novel, sensitive, low-cost detection method, EvaGreen-based real-time PCR assay for the detection of SAV. This assay exhibited high specificity for SAV1, SAV2, and SAV5 and was able to detect SAV at concentrations as low as 1.5 × 101 copies, making them more sensitive than the approved conventional RT-PCR method (detection limit, 1.5 × 106 copies). Assessment of infected fish samples showed that the sensitivity of EvaGreen-based assay was higher than previously developed SYBR Green assay (227 assay). Thus, we report that the EvaGreen real-time PCR assays is an economical alternative diagnostic method for the rapid detection of SAV1, SAV2, and SAV5 infection, providing improved technical support for the clinical diagnosis and epidemiological investigation of SAV.
Source: Molecular and Cellular Probes - Category: Molecular Biology Source Type: research