Nsp9基因实时荧光定量PCR检测方法的建立及其在感染细胞过程中表达量的变化

doi:10.16431/j.cnki.1671-7236.2016.10.004

Abstract

Abstract:

The study was conducted to establish a quantitative Real-time PCR method for PRRSV Nsp9 gene rapid detection and study its expression in PRRSV infected cells.A pair of specific primers targeted to Nsp9 of PRRSV was designed and a Real-time PCR method based on SYBR Green Ⅰ fluorescent was developed for the quantization of PRRSV.The melting curve analysis using SYBR Green Ⅰ dye showed one specific peak,and no primer dimers peak was observed.No amplification was detected from HEV,SIV and PRV samples by this method.There was good reproducibility and low variation coefficient.The quantitative Real-time PCR method developed in this study would be useful for rapid laboratory diagnosis and epidemiology investigation of PRRSV.During the process of virus infecting cells,the expression level of Nsp9 increased gradually,and it got the highest at 36 h.This study laid the theoretical basis to explore the law of virus replication and clinical vaccine production.

Key words: PRRSV; SYBR Green Ⅰ; quantitative Real-time PCR; Nsp9 gene

CLC Number:

S852.65

ZHAO Meng-meng, FENG Song-lin, WANG Wen-jia, XING Xing, FENG Jia-ping, ZHANG Gui-hong. Establishment of the Quantitative Real-time PCR Method for PRRSV Nsp9 Gene Rapid Detection and Its Expression in PRRSV Infected Cells[J]. , 2016, 43(10): 2534-2540.

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Establishment of the Quantitative Real-time PCR Method for PRRSV Nsp9 Gene Rapid Detection and Its Expression in PRRSV Infected Cells

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