Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Highly Sensitive Detection of Low-Abundance White Spot Syndrome Virus by a Pre-Amplification PCR Method

  • Pan, Xiaoming (College of Food Science and Engineering, Ocean University of China) ;
  • Zhang, Yanfang (College of Food Science and Engineering, Ocean University of China) ;
  • Sha, Xuejiao (College of Food Science and Nutritional Engineering, China Agriculture University) ;
  • Wang, Jing (College of Food Science and Engineering, Ocean University of China) ;
  • Li, Jing (College of Food Science and Engineering, Ocean University of China) ;
  • Dong, Ping (College of Food Science and Engineering, Ocean University of China) ;
  • Liang, Xingguo (College of Food Science and Engineering, Ocean University of China)
  • Received : 2016.06.24
  • Accepted : 2016.12.09
  • Published : 2017.03.28
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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 ${\sim}10^{10}$ magnitude 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.

Keywords

References

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