Reverse transcription polymerase chain reaction (RT-PCR) was used for the detection of classical swine fever virus (CSFV) in blood and tissue samples of field cases and experimentally inoculated pigs. The distribution...Reverse transcription polymerase chain reaction (RT-PCR) was used for the detection of classical swine fever virus (CSFV) in blood and tissue samples of field cases and experimentally inoculated pigs. The distribution of CSFV in different organ samples showed some discrepancies in infected pigs. Four weaner pigs were inoculated with C-strain vaccine virus, then samples of spleen, tonsil, lung, mesenteric lymph node, kidney and brain were collected after slaughter and tested for E2 and NS5B genes using one-step RT-PCR and nested RT-PCR. Using the same method, 12 field cases were simultaneously studied. A discrepancy of CSFV in different samples was found upon detecting the target gene. The most reliable diagnostic organs were spleen and tonsil, and the nested RT-PCR assay provided a highly sensitive and specific method with comparable performance to the one-step RT-PCR assay.展开更多
Maedi-visna virus（MVV） is an ovine lentivirus that is widespread in many countries worldwide.Both clinical and subclinical MVV infections cause substantial economic losses.MVV infection in live sheep is usually diag...Maedi-visna virus（MVV） is an ovine lentivirus that is widespread in many countries worldwide.Both clinical and subclinical MVV infections cause substantial economic losses.MVV infection in live sheep is usually diagnosed serologically,with antibody-positive sheep being regarded as infected.There have been few reports of maedi-visna in China,with no detailed epidemic analysis of MVV infection in ovine herds.In order to investigate the seroprevalence and spatial distribution of maedi-visna among ovine flocks in China,a total of 672 serum samples were collected from different ovine flocks in 12 regions（provinces,autonomous regions or municipalities） of China in 2011,and serum antibody levels were determined using a commercial ELISA Kit.This study represents the first investigation of the seroepidemiology of maedi-visna in China,indicating a circulation of MMV among sheep.展开更多
We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constr...We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.展开更多
基金supported by the grants from the National"863" Programme (2006AA10A2041)Development Programme of Science and Technology,Chengguan District of Gansu province (08-5-4)
文摘Reverse transcription polymerase chain reaction (RT-PCR) was used for the detection of classical swine fever virus (CSFV) in blood and tissue samples of field cases and experimentally inoculated pigs. The distribution of CSFV in different organ samples showed some discrepancies in infected pigs. Four weaner pigs were inoculated with C-strain vaccine virus, then samples of spleen, tonsil, lung, mesenteric lymph node, kidney and brain were collected after slaughter and tested for E2 and NS5B genes using one-step RT-PCR and nested RT-PCR. Using the same method, 12 field cases were simultaneously studied. A discrepancy of CSFV in different samples was found upon detecting the target gene. The most reliable diagnostic organs were spleen and tonsil, and the nested RT-PCR assay provided a highly sensitive and specific method with comparable performance to the one-step RT-PCR assay.
基金supported by the fund of China Agriculture Research System(CARS-39)
文摘Maedi-visna virus（MVV） is an ovine lentivirus that is widespread in many countries worldwide.Both clinical and subclinical MVV infections cause substantial economic losses.MVV infection in live sheep is usually diagnosed serologically,with antibody-positive sheep being regarded as infected.There have been few reports of maedi-visna in China,with no detailed epidemic analysis of MVV infection in ovine herds.In order to investigate the seroprevalence and spatial distribution of maedi-visna among ovine flocks in China,a total of 672 serum samples were collected from different ovine flocks in 12 regions（provinces,autonomous regions or municipalities） of China in 2011,and serum antibody levels were determined using a commercial ELISA Kit.This study represents the first investigation of the seroepidemiology of maedi-visna in China,indicating a circulation of MMV among sheep.
基金supported by the National Programs for High Technology Research and Development of China (2006AA10A204)the Gansu Key Technologies R&D Program(ZGS-052-A41-0006-03)the Programs for Director Fund of Lanzhou Veterinary Research Institute
文摘We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.
