Citation: WAN Chao, WEN Guo-yuan, PAN Zi-shu, ZHANG Chu-yu, XIONG Zhong-liang, YANG Jun, AI Di-yun. Development of Fluorescent Quantitative PCR Method to Rapidly Detect Pseudorabies Virus .VIROLOGICA SINICA, 2006, 21(5) : 485-489.

Development of Fluorescent Quantitative PCR Method to Rapidly Detect Pseudorabies Virus

  • Available online: 20 September 2006
  • A fluorescent quantitative PCR (FQ-PCR) method based on sequences of the conserved gE of the PRV genome was established and evaluated after selection of optimal reaction conditions. A pair of primers and a fluorescent TaqMan probe specific for gE gene were designed and used. We compared the FQ-PCR assay to conventional virus culture techniques and PCR test. According to our results, the dynamic range of the FQ-PCR assay is between 1.0×102 copies/L and 1.0×107copies/L, and the lowest DNA concentration of detection is 1.0×102 copies/L, which is 10 fold better than regular PCR and also avoids non-specific amplification occasionally seen in regular PCR. We tested 66 specimens from different pig farms in Hubei, Henan and Gansu in 2005 by the FQ-PCR assay and isolates of PRV were found in 42 samples (63.6%). In conclusion, the FQ-PCR method is rapid, sensitive, specific and accurate and can be used to rapidly detect wild type PRV from pig’s tissues and respirator specimens.

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    Development of Fluorescent Quantitative PCR Method to Rapidly Detect Pseudorabies Virus

    • 1. 1.College of Life Sciences, State Key Laboratory of Virology, Wuhan University, Wuhan 430072, China
    • 2. Institute of Animal Husbandry and Veterinary Science, Hubei Academy of Agricultural Science, Wuhan 430209 , China

    Abstract: A fluorescent quantitative PCR (FQ-PCR) method based on sequences of the conserved gE of the PRV genome was established and evaluated after selection of optimal reaction conditions. A pair of primers and a fluorescent TaqMan probe specific for gE gene were designed and used. We compared the FQ-PCR assay to conventional virus culture techniques and PCR test. According to our results, the dynamic range of the FQ-PCR assay is between 1.0×102 copies/L and 1.0×107copies/L, and the lowest DNA concentration of detection is 1.0×102 copies/L, which is 10 fold better than regular PCR and also avoids non-specific amplification occasionally seen in regular PCR. We tested 66 specimens from different pig farms in Hubei, Henan and Gansu in 2005 by the FQ-PCR assay and isolates of PRV were found in 42 samples (63.6%). In conclusion, the FQ-PCR method is rapid, sensitive, specific and accurate and can be used to rapidly detect wild type PRV from pig’s tissues and respirator specimens.

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