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Rapid detection of goose hemorrhagic polyomavirus using TaqMan quantitative real-time PCR
Chunhe Wan1, Longfei Cheng1, Guanghua Fu1
1Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention/Fujian Animal Diseases Control Technology Development Center, Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences, Fuzhou 350013, People's Republic of China.
Abstract:
Due to low doses of infection, an efficient and sensitive virus detection method is necessary to detect low amounts of goose hemorrhagic polyomavirus (GHPV). In this study, we have developed a TaqMan real-time PCR (qPCR) specific assay for the detection of GHPV. Specificity assay showed no cross-reactions with other common waterfowl viruses. The standard curve had a linear correlation of 0.997 and efficiency of 99% between the cycle threshold value and the logarithm of the plasmids copy number. The possible lowest detectable concentration was 35.4 copies/μl; 100 times more sensitive than conventional PCR (detection limit, 3.54 × 103 copies/μl). Domestic Jinyun Sheldrakes ducks and their embryonated eggs were found positive of GHPV infection which provides evidence of possible vertical transmission of GHPV.
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