Development Of PCR For Detection Of Goose Parvovirus And Molecular Epidemiological Investigation Of Goose Parvovirus In Jianghuai Areas

1. Development of PCR for Detection of Goose ParvovirusGosling plague is caused by gooes parvovirus, it is a septic infectious diseases in gosling and Muscovy duckling. With the development of the goose industry, Gosling plague has made huge economic loss and serious threat to goose industry. In this study, we successfully established a PCR method for the detecting GPV. At the same time we investigated the molecular epidemiology of GPV in Jianghuai areas. Our results would be benefit to the development of diagnostic methods for detecting GPV and reducing economic losses in the future.One pair of primers was designed according to the conservative VP3region sequences of goose parvovirus B strains in GenBank. We established a PCR method for the detection of GPV. Using the PCR assay, the specific fragment of776bp was amplified from DNA sample of the GPV only, not from ALV, AIV, DTMUV, EDS76, REV, MDV and goose pathogenic Escherichia coli. The sensitivity of the PCR was4.7ng/50μl for the GPV nucleic acids. All the specific fragments were amplified by this method from41samples (including liver and intestine) of goslings infected with GPV clinically. The positive rate was100%. At the same time32strains GPV was isolated from the41samples, positive rate was78.0%. The resultes indicating that the PCR assay established for detecting GPV was sensitive and specific, which had potential clinical value.2. Molecular Evolution of Goose Parvovirus Isolated in Jianghuai AreasTo investigate the prevalence of Goose Parvovirus in Jianghuai areas, a total of41samples of suspected goslings infected with GPV including liver and intestine were collected from2012to2013in Jiangsu province, and33strains of GPV were isolated and identified. A pair of primers was designed to amplify the VP3genes. Sequences analysis demonstrated that the VP3gene of the23strains consisted of1605bp and encoded534amino acids. It showed that 91.3%-99.9%homology at the DNA level and94.1%-99.6%homology at the amino acid level to the reference strains in GeneBank. The phylogenetic analysis based on the VP3gene sequences revealed that the GPV has two main subgroups, one GPV strain from Muscovy ducks which named DY belonged to subgroup1and most of other GPV strains isolated in this study were clustered in subgroup2. The absence of the deduced514-517NETG glycosylation site in VP3region may explain the host specificity of the GPV.3. Complete gene Sequence Analysis and Molecular Evolution of Goose ParvovirusTwo GPV strains were isolated from suspected GPV infection in clinic, designated as GPV-CZM and GPV-SHFX120619. The complete DNA sequences of GPV-CZM and GPV-SHFX120619were amplified by polymerase chain reaction (PCR). The genome of the GPV-CZM has long5106nucleotides (nt), just as the same as strain B. The results revealed that the homology of full nucleotide sequences was93.1%and98.9%with the reference strains B and GDaGPV; The genome of the GPV-SHFX20120619(GenBank:KC478066) has long5050nucleotides (nt),56bp deletion was found in ITR sequence compared with strain B, but same to the strain82-0321. The results revealed that the homology of full nucleotide sequence was98.2%and99.7%with the reference strains B and82-0321respectively.