Real-time PCR assay for universal detection and quantitation of all five species of fowl adenoviruses (FAdV-A to FAdV-E).
作者： Ayse Günes , Ana Marek , Beatrice Grafl , Evelyn Berger , Michael Hess
DOI: 10.1016/J.JVIROMET.2012.04.005
关键词: Real-time polymerase chain reaction 、 Viral shedding 、 Virology 、 Serotype 、 Serial dilution 、 DNA 、 Gene 、 Fowl 、 Biology 、 Molecular biology 、 Polymerase chain reaction
摘要: The present study describes the development of a SYBR Green based real-time polymerase chain reaction (real-time PCR) for detection and quantitation all fowl adenovirus (FAdV) species. Primers were designed on conserved nucleotide sequences within 52K gene. Ten-fold serial dilutions vector DNA used as standard quantitation. PCR had an efficiency 98%, regression squared value 0.999 showed range 6.73-6.73×10(8) copies FAdV per reaction. assay was highly specific FAdVs exact 5 species (FAdV-A to FAdV-E) could be demonstrated. It shown, that twelve serotypes (FAdV-1 8a, 8b 11) detectable quantifiable. Other viral genomes well uninfected chicken embryo liver (CEL) cells did not produce positive signal. Cloacal swabs taken during animal experiment, which performed with Shedding investigated in cell culture, by conventional developed PCR. found more sensitive than culture Detection different type samples possible new
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