The establishment and application of isothermal cross-priming amplification techniques in detecting penaeid shrimp white spot syndrome virus.
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°C and 60 min, respectively. The detection limit of the WSSV-CPA assay was as low as 10 copies μl(-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.


CONCLUSIONS
White spot syndrome, caused by white spot syndrome virus (WSSV), is the major disease threatening the shrimp aquaculture industry and leads to tens of billion dollars of economic losses in the world each year. This study established a CPA-based method for detecting WSSV, which is rapid, sensitive and specific. It is anticipated that this novel assay will be instrumental for diagnosis and surveillance of WSSV.
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, China; Shanghai Ocean University, Shanghai, China; National Laboratory for Marine Science and Technology, Qingdao, China.
H-L
HL
Yang
J
J
Huang
B
B
Yang
F
F
Liu
Q-L
QL
Zhang
D000818:Animals; D017756:Aquaculture; D017931:DNA Primers / Q000235:genetics; D005814:Genes, Viral; D025202:Molecular Diagnostic Techniques; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D046848:White spot syndrome virus 1 / Q000235:genetics*
D017931:DNA Primers
Cross-priming amplification; detect; polymerase chain reaction; white spot syndrome virus