A sensitive and specific SYBR Green-based qPCR assay for detecting scale drop disease virus (SDDV) in Asian sea bass
| Autor: | Chuenchit Boonchird, Siripong Thitamadee, Saengchan Senapin, Ha Thanh Dong, Molruedee Sonthi, Sukhontip Sriisan |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Disease
Aquatic Science Biology Real-Time Polymerase Chain Reaction Megalocytivirus Virus Fish Diseases 03 medical and health sciences chemistry.chemical_compound Plasmid Animals Sea bass Ecology Evolution Behavior and Systematics 030304 developmental biology Subclinical infection 0303 health sciences Aquatic animal 04 agricultural and veterinary sciences biology.organism_classification Virology Iridoviridae chemistry 040102 fisheries 0401 agriculture forestry and fisheries Bass DNA |
| Zdroj: | Diseases of Aquatic Organisms. 139:131-137 |
| ISSN: | 1616-1580 0177-5103 |
| DOI: | 10.3354/dao03484 |
| Popis: | Scale drop disease virus (SDDV) is a megalocytivirus known to cause disease in Asian sea bass in Southeast Asia. To support SDDV diagnosis and surveillance, we report on a sensitive and specific SYBR Green qPCR assay. The qPCR primers were designed to target a 135 bp fragment of the SDDV ATPase gene. The optimized SDDV qPCR assay reliably detected 2 copies of a plasmid dsDNA control and did not cross-amplify DNA to any of 12 viral or bacterial pathogens commonly found in aquatic animals. When assessed with 86 field samples, the assay detected SDDV in DNA extracted from each of 34 scale drop disease-affected fish collected from 5 affected farms. The qPCR also detected SDDV in DNA from 30 of 52 overtly healthy fish collected from 9 farms where SDDV had not been detected previously, using a semi-nested conventional PCR. The higher sensitivity of our SDDV qPCR assay can thus be useful in detecting fish with subclinical/chronic infections. However, the qPCR showed that SDDV DNA loads varied from 8.0 × 102 to 6.8 × 104 viral DNA copies per 200 ng DNA template among the 8 organ tissue types sampled from 3 diseased fish. In circumstances requiring SDDV to be detected unequivocally in subclinical carriers with lower-level infection, qPCR testing of more than one type of tissue is advisable. |
| Databáze: | OpenAIRE |
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