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Highly sensitive detection of low-abundance of white spot syndrome virus by a pre-amplification PCR method.

Highly sensitive detection of low-abundance of white spot syndrome virus by a pre-amplification PCR method.
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Pan X, Zhang Y, Sha X, Wang J, Li J, Dong P, Liang X,


Pan X, Zhang Y, Sha X, Wang J, Li J, Dong P, Liang X, (click to view)

Pan X, Zhang Y, Sha X, Wang J, Li J, Dong P, Liang X,

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Journal of microbiology and biotechnology 2016 12 14() doi 10.4014/jmb.1606.06053
Abstract

White spot syndrome virus (WSSV) is a major threat to shrimp farming industry and so far there’s no effective therapeutic way to it, thus early diagnostic of WSSV is of great importance. However, at the early stage of infection the extremely low-abundance of WSSV DNA challenged the detection sensitivity and accuracy of PCR. To effectively detect low-abundance of 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, ten copies of target WSSV gene were detected from ~10¹⁰ magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of universal primer. The efficiency of pre-amp PCR was validated by nested-PCR and quantitative PCR (qPCR), 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 diagnostic of WSSV, meanwhile it has potential to be applied in other low-abundance sample detection cases.

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