Sequence Analysis Of Ducv In Guangxi And The Development Of The Molecular Biological Method Of Ducv

DuCV is one of the new virus found in recent years, it was first discovered and finished the gene sequencing in2003. Soike and someone found DuCV basically make the duck slow growth, feathers messy, weight loss and other symptoms in clinical. More important is that the immune system of birds can be infected, leading to the immune suppression. DuCV make a great affect to the economic benefits of breeding.This study identified the DuCV of Guangxi isolate, and then cloned and sequenced the entire genome. Compared with the entire genome of DuCV on GenBank, the result shows that the homology reach up to97%compared with the DuCV in Taiwan. The homology reach up from76%to79%compared with the DuCV in some provinces in China、North America and Europe.Now there are traditional diagnosis methods and diagnostic molecular techniques to detect DuCV. Loop-mediated isothermal amplification is a kind of new Molecular diagnosis technology. The results showed that the assay had no cross-reaction with H9subtype AIV Gosling Plague virus, Duck Plague virus, Duck Hepatitis Virus, Duck Paramyxo virus and other grallatorial pathogens. The assay was performed in water bath within one hour and the amplification result was visualized by by inspecting the white sediment and the green colour.. The detection limit of LAMP assay was1fg, which was1000-fold higher than that of routine PCR. These results suggest that the LAMP assay is a simple and specific method for rapid detection of DuCV in field conditions.Besides, SYBR Green I real-time PCR is a kind of new molecular diagnosis methods. Results shows that it has become an impretant detection method of pathogeny because of the high specificity, high sensitivity, reaction quickly and the low cost. Sensitivity analysis showed that the developed SYBR Green I real-time PCR could detect1×102template copies of plasmid DNA in one reaction. The assay exhibited specificity as all negative controls and other avian pathogens,such as H9subtype AIV, Gosling Plague virus, Duck Plague virus, Duck Hepatitis Virus and other grallatorial pathogens showed negative results. And the results showed that the assay have good repeatability.As a result of the sensitivity and specificity of the assay with a rapid and simple procedure,the real-time PCR will be useful for the clinical diagnosis of DuCV infection.A multiplex polymerase chain reaction was optimized to simultaneously detect two pathogenic of DHV and DuCV in this article.DuCV and DHV are very common in ducks. The two virus caused huge losses because of widely prevalent, rapidly onset, highly death rate and mixed infection. It will make great losses if ducks infected both DuCV and DHV. A multiplex polymerase chain reaction is a special kind of PCR, two pathogeny can be identified in a reaction. Multiplex polymerase chain reaction has a very high application value in clinic. It turned out that as little as56pg of DHV RNA(569bp) and6pg of DuCV DNA(245bp) was detected using gel electrophoresis in this multiplex PCR. And no specific band was amplified from other avian pathogenic virus and bacteria.This study explores to provide the theory basis and the technical support for DuCV.