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Virologica Sinica, 21 (5) : 485-489, 2006
Research Article
Development of Fluorescent Quantitative PCR Method to Rapidly Detect Pseudorabies Virus
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
 Correspondence:
(376.60KB)  
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/µL and 1.0×107copies/µL, and the lowest DNA concentration of detection is 1.0×102 copies/µL, 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.
  Published online: 20 Sep 2006
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