Journal
POULTRY SCIENCE
Volume 99, Issue 12, Pages 6586-6592Publisher
ELSEVIER
DOI: 10.1016/j.psj.2020.09.077
Keywords
Novel goose astrovirus; EvaGreen dye; qLAMP; PCR; ORF1a
Categories
Funding
- National Key Research and Development Program of China [2018YFD0501506]
- Higher Education Support Program of Youth Innovation and Technology of Shandong Province, China [2019KJF022]
- China Agriculture Research System [CARS-42-19]
- Shandong Double Tops Program
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In November 2017, a severe infectious disease that devastated the major goose-producing regions in China was found to be caused by a novel goose astrovirus (N-AstV). The objective of this study was to develop a quantitative loop-mediated isothermal amplification (qLAMP) assay for the rapid diagnosis of N-AstV characterized with gout, hemorrhage, and swellings of the kidneys. A set of 4 specific primers, 2 inner and 2 outer primers, targeting the ORF1a gene of N-AstV were designed for the assay which could be completed within 60 min at 65 degrees C in a water bath or on a real-time PCR instrument for quantitative analysis. The qLAMP assay showed a high sensitivity with a detection limit of 1 x 10(1) copies of the target DNA/mu L. There were no cross-reactions with other viruses, and the reproducibility of the assay was confirmed in intrasensitivity and intersensitivity assay tests with variability ranging from 0.61 to 2.21%. The results indicated that the qLAMP assay for N-AstV was a simple, accurate, rapid, sensitive, and specific, especially useful for field detection.
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