茶尺蠖核型多角体病毒荧光定量PCR检测方法的建立

doi:10.13305/j.cnki.jts.2010.03.009

Abstract

Abstract: A pair of primer was designed according to the Ectropis oblique nucleopolyhedrovirus (EoNPV) genome DNA sequence. Virus DNA was extracted from the tea looper larvae, Ectropis oblique which were infected with the EoNPV. The DNA fragment was amplified, then cloned into pMD18-T vector and transferred into E. coli TG1. A single clone was selected and sequenced, and the extracted recombinant plasmid DNA was used as a positive quantitative template to establish a standard curve. The standard curve showed a linear relationship between cycle threshold (CT) and template concentration ranging from 10³~10⁸ copies/µL with a correlation coefficient of 0.989, and the quantitative PCR was more repeatable and specific than traditional PCR. The fluorescent quantitative PCR method for detecting EoNPV was developed, providing a basis for the research of EoNPV replicating in the host body and quantitative detection of the EoNPV in biopesticide products.

Key words: ectropis oblique, nucleopolyhedrovirus, fluorescent quantitative PCR, detection

CLC Number:

DU Jun-li, ZHANG Chuan-xi, XIAO Qiang, FU Jian-yu, YIN Kun-shan. Development of the Fluorescent Quantitative PCR for Detection of EoNPV in Ectropis Oblique[J]. Journal of Tea Science, 2010, 30(3): 203-207.

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Development of the Fluorescent Quantitative PCR for Detection of EoNPV in Ectropis Oblique

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