鸭坦布苏病毒和鸭瘟病毒二重荧光定量RT-PCR方法的建立

Abstract

Abstract: The study was conducted to establish the duplex Real-time PCR assay for detecting both duck tembusu virus (DTMUV) and duck plague virus (DPV). According to the sequences of DTMUV E gene and DPV UL6 gene in GenBank, two sets of specific oligonucleotide primers for DTMUV and DPV along with two TaqMan probes were designed. The duplex Real-time PCR assay was developed through optimization of reaction conditions and validation of specificity, sensitivity and repetitiveness of the method. The sensitivity of the assay were both 100 template copies for DTMUV and DPV. There was no specific bands of the same sizes were amplified from other duck pathogens, such as duck Newcastle disease virus, duck hepatitis virus, muscovy duck parvovirus, duck circovirus, H9 subtype avian influenza virus, egg drop syndrome virus. This duplex Real-time RT-PCR assay is a sensitive, quick, specific and quantitative test for detection of DTMUV and DPV, and will be useful for the control of these viruses in ducks.

Key words: duck tembusu virus; duck plague virus; duplex Real-time RT-PCR

ZHANG Yan-fang, XIE Zhi-xun, XIE Li-ji, LIU Jia-bo, FAN Qing, PANG Yao-shan, LUO Si-si, DENG Xian-wen, XIE Zhi-qin. Development of Duplex Real-time RT-PCR Assay for Detection of Duck Tembusu Virus and Duck Plague Virus[J]. .

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Development of Duplex Real-time RT-PCR Assay for Detection of Duck Tembusu Virus and Duck Plague Virus

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