Journal
LETTERS IN APPLIED MICROBIOLOGY
Volume 59, Issue 2, Pages 200-206Publisher
WILEY
DOI: 10.1111/lam.12267
Keywords
Cross-priming amplification; detect; polymerase chain reaction; white spot syndrome virus
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Funding
- Special Fund for Agro-scientific Research in the Public Interest [201103034]
- China Agriculture Research System [CARS-47]
- Construction Programme for 'Taishan Scholarship' of Shandong Province of China
- Special Scientific Research Funds for Central Nonprofit Institutes, Chinese Academy of Fishery Sciences [2013A0601]
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White spot syndrome virus (WSSV) is one of the main pathogens seriously threatening penaeid shrimp farming in the world. This study designed and synthesized five specific primers based on the conserved sequences of WSSV genome, optimized the reaction system and conditions, and finally established a cross-priming amplification (CPA)-based detection method for WSSV (WSSV-CPA). The results indicated that the optimized reaction temperature and time of WSSV-CPA were 63 degrees C and 60 min, respectively. The detection limit of the WSSV-CPA assay was as low as 10 copies mu l(-1) and shared same sensitivity with the WSSV-qPCR assay. Due to dispensing with expensive thermal cycler, time- and cost-saving, and ease of use in field, it is anticipated that the WSSV-CPA method developed in this study will be instrumental for the diagnosis and surveillance of WSSV.
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