Rapid and Sensitive qRT-PCR Assay for Fish VHSv Detection

What is it about?

The article presents a novel, easy, rapid, and highly sensitive real-time quantitative reverse transcription PCR (qRT-PCR) assay for detecting and quantifying the Viral Hemorrhagic Septicemia virus (VHSv) in fish species. The assay incorporates synthetic competitive template internal standards for quality control and has a linear dynamic range that spans seven orders of magnitude. Compared to traditional cell culture diagnostics, the new assay is more sensitive and accurate, detecting VHSv levels ranging from 6 to 6,000,000 molecules per 10^6 actb1 molecules. The assay detects all known VHSv strains and substrains, including the virulent IVb strain responsible for outbreaks in the Great Lakes. It also provides valuable information on the viral load associated with clinical signs of disease in infected fish. This diagnostic tool will aid in the early detection, prevention, and control of VHSv, a serious fish pathogen causing significant economic loss in the aquaculture and fishery industries.

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Why is it important?

This research is important for several reasons: Disease control: Rapid, sensitive, and accurate diagnostic methods are crucial for the early detection and control of viral diseases in fish populations, particularly those that cause significant economic and ecological impacts like VHSv. Cost-effective solutions: The new qRT-PCR assay is more economical and efficient than traditional cell culture diagnostics, offering a more practical and accessible solution for detecting and quantifying VHSv in fish populations. Detection of all known VHSv variants: The assay is specific to VHSv and can detect all known variants of the virus, helping researchers understand the prevalence and distribution of the virus in different geographic regions. Improved diagnostic tools: The development and implementation of this highly sensitive and accurate qRT-PCR assay contribute to the ongoing efforts to improve diagnostic tools for fish diseases, ultimately benefiting fisheries management and conservation efforts. Application in other fields: The methodology and principles behind this qRT-PCR assay can potentially be applied to develop diagnostic tools for other pathogens, both in the fish and other animal industries, and in human health. Key Takeaways: 1. The new qRT-PCR assay is a highly sensitive, accurate, and rapid diagnostic tool for detecting and quantifying VHSv in fish populations, surpassing traditional cell culture diagnostics and other published PCR-based assays. 2. The assay is specific to VHSv and can detect all known variants of the virus, aiding in the diagnosis of viral infections and understanding the spread of the virus in different geographic regions. 3. The inclusion of synthetic competitive template internal standards in the assay improves its accuracy and helps overcome the limitations of other published PCR-based tests, which often resulted in high numbers of false negatives. 4. The qRT-PCR assay is cost-effective and efficient, offering a practical solution for detecting and monitoring VHSv in fish populations, aiding in disease control and management efforts.