title: Highly Sensitive Detection of Low-Abundance White Spot Syndrome Virus by a Pre-Amplification PCR Method.
abstract: White spot syndrome virus (WSSV) is a major threat to the shrimp farming industry and so far there is no effective therapy for it, and thus early diagnostic of WSSV is of great importance. However, at the early stage of infection, the extremely low-abundance of WSSV DNA challenges the detection sensitivity and accuracy of PCR. To effectively detect low-abundance WSSV, here we developed a pre-amplification PCR (pre-amp PCR) method to amplify trace amounts of WSSV DNA from massive background genomic DNA. Combining with normal specific PCR, 10 copies of target WSSV genes were detected from ~1010magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of the universal primer. The efficiency of the pre-amp PCR was validated by nested-PCR and quantitative PCR, and pre-amp PCR showed higher efficiency than nested-PCR when multiple targets were detected. The developed method is particularly suitable for the super early diagnosis of WSSV, and has potential to be applied in other low-abundance sample detection cases.
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, P.R. China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Pan
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, P.R. China.|School of Life Sciences, Inner Mongolia University, Hohhot 010000, P.R. China.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Food Science and Nutritional Engineering, China Agriculture University, Beijing 100083, P.R. China.
authors.forename: Xuejiao
authors.initials: X
authors.lastname: Sha
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, P.R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, P.R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, P.R. China.
authors.forename: Ping
authors.initials: P
authors.lastname: Dong
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, P.R. China.
authors.forename: Xingguo
authors.initials: X
authors.lastname: Liang
mesh_terms: D000820:Animal Diseases / Q000175:diagnosis / Q000821:virology; D000818:Animals; D005814:Genes, Viral; D016679:Genome, Viral; D060885:Multiplex Polymerase Chain Reaction; D033561:Penaeidae / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D046848:White spot syndrome virus 1 / Q000235:genetics*
keywords: White spot syndrome virus; detection; high efficiency; multiplex; pre-amplification PCR