This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of...This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of which 2 samples were genetically close to the X7443 strain (Nigeria 75/1) of lineage I, and 3 samples close to the strain AY560591 (Sungri96) of linage IV with 96.6%、97.3%、97.6% and 98% nucleotide sequence homogeneity respectively, based on partial sequencing of the F gene from 5 samples and complete sequencing of the N/M/F/H genes from one sample. This study suggested that there are at least 2 origins of PPRV in China.展开更多
This experiment was conducted to diagnose Peste des Petits Ruminants based on the eukaryotically-expressed PPRV nucleoprotein through an indirect ELISA. The full-length PPRV nucleoprotein gene was obtained from viral ...This experiment was conducted to diagnose Peste des Petits Ruminants based on the eukaryotically-expressed PPRV nucleoprotein through an indirect ELISA. The full-length PPRV nucleoprotein gene was obtained from viral genome RNA by RT-PCR. The amplified fragments were cloned into baculovirus donor vectors pFastHTA of the Bac-to-Bac system. These recombinant plasmids, pFastHTA-PPRV-N, were transformed into DH10Bac host bacteria to obtain recombinant shuttle plasmids, pBacmid-PPRV-N. Recombinant baculovirus, Bacmid-PPRV-N, was generated for expression of the PPRV nucleoprotein by transfecting recombinant pBacmid-PPRV-N with Lipofectamine 2000 into Sf21 insect cells. The efficient expression of PPRV Nucleoprotein by baculovirus in Sf21 cells was verified by SDS-PAGE and Western blot. An indirect ELISA was developed using recombinant PPRV nucleoprotein as the coating antigen. 37 goat sera from an epidemic area in Tibet and 92 goat sera from a non-infected area in Qinghai Province were simultaneously detected by the indirect ELISA, developed here, and the international standard cELISA kit. The sensitivity and specificity of the indirect ELISA was 100% and 96.2% compared with the cELISA kit. The coincidence rate of the two methods was 96.9%. The results demonstrated that the indirect ELISA established in this study works well for diagnosis of PPR.展开更多
Peste ties petits ruminants is a kind of acute eontagious disease infecting goats anti sheep. In this study, antibtly monitoring and tracking of healthy goat and sheep immunized by peste des petits ruminants vaccine i...Peste ties petits ruminants is a kind of acute eontagious disease infecting goats anti sheep. In this study, antibtly monitoring and tracking of healthy goat and sheep immunized by peste des petits ruminants vaccine in Changping District of Beijing City were conducted, aiming at providing a reference for the devel- opment of effective immunization procedures.展开更多
Peste des petits ruminant (PPR) is a contagious disease of small ruminants caused by a virus that belongs to the genus Morbillivirus of the family Paramyxoviridae. This study aimed to determine the seroprevalence of P...Peste des petits ruminant (PPR) is a contagious disease of small ruminants caused by a virus that belongs to the genus Morbillivirus of the family Paramyxoviridae. This study aimed to determine the seroprevalence of PPR disease in sheep and goats and its associated risk factors in Kassala State, Eastern Sudan. Across sectional study was conducted during the period from 30th August to 25th November 2015. The study was carried out using a structured questionnaire survey and a total of 918 blood samples were collected from apparently healthy unvaccinated sheep and goats in different localities in State of Kassala. A total of 546 sheep and 372 goats were tested for specific antibodies to nucleoprotein (NP) by competitive enzyme linked immunosorbent assay (cELISA). The apparent overall prevalence of PPR antibodies in Kassala was 58.2% while the true prevalence was calculated to be 61.3%. The apparent prevalence in sheep and goats was 68.1% and 43.5% respectively. Univariate analysis showed that the risk factors had significant associations with a cELISA positive status: locality, species, age, breed, husbandry system, housing mode, animals movement (p = 0.000) and animals sharing pasture and water (p = 0.003), while sex and newly introduced animals were not significant risk factors (p = 0.771) (p = 0.050) respectively. Factors found that significantly associated (p < 0.05) with increased odds of being cELISA positive in multivariate analysis were localities, species, age and newly introduced animals. The prevalence differed between localities and was the highest in the River Atbara (84.0%) locality, whereas it was lowest in Delta North (29.0%). No significant difference was observed among the sexes. However, the prevalence differed in different age groups and was 52.25% in animals of less than six months old;49.3% were between seven months and two years old and 65.5% were above two years old. In different husbandry systems, the prevalence was 47.9%, 73.0% and 49.2% in intensive, open grazing and pastoral systems respectively. Housing type effects were also observed;the highest prevalence was in animals housed in metal fence (83.3%). The movement pattern showed significant effect, where the prevalence was the highest (81.3%) in animals that move inter-states/inter-localities. It is concluded that the disease is endemic in Kassala State, high prevalent in sheep and goats, posing a threat to animal exportation, and may have a serious economic influence. Owners and herders should compulsorily vaccinate their animals yearly and animals should be investigated periodically for implementation of crucial eradication program.展开更多
The present study was carried out between April 2015 and January 2016 to estimate the sero-prevalence and identify the risk factors of the peste des petits ruminants (PPR) in Cameroon. A total of 269 herds randomly sa...The present study was carried out between April 2015 and January 2016 to estimate the sero-prevalence and identify the risk factors of the peste des petits ruminants (PPR) in Cameroon. A total of 269 herds randomly sampled across the country have been studied and 1622 samples of serum have been levied on the sheep and goat. The c-ELISA has been studied in order to detect the presence of antibodies in small ruminants like an indicator of exposition to PPRV. The results revealed the circulation of PPRV in the country with a total sero-prevalence of 39% [95%CI;37 - 41] and a sero-prevalence of 63.2% [95%CI;57.2 - 69.2] at the herd level. Sero-prevalence was variable in the ten regions ranging from 7% [95% CI;6.2 - 8.4] to 73% [95% CI;62 - 84] with the northern zone (Adamawa, North and Far-North) having 52.3% [95% CI;37 - 60] and southern zone (including the remaining seven regions) recording 29% [95% CI;11 - 57]. Similarly, it was higher in animals found in urban/peri-urban areas than in rural areas with prevalence ratio of 2.9 [95% CI 2.54 - 3.4;p < 0.001] <em>i.e. </em>3 times more, 1.6 [95% CI 1.36 - 1.90;p < 0.001] <em>i.e.</em> 1.6 times more, and 5.02 [95% CI 3.91 - 6.85;p < 0.001] <em>i.e.</em> 5 times more at national level, in the northern zone and in the southern area, respectively. Five risk factors have been identified: the breeding environment, introduction of new animals into the herds, gathering of animals for pasture and watering, wandering and transhumance. The breeding area appeared to be the most important risk factor associated with disease exposure. The control measures for the eradication of this disease must take into account the epidemiological situation, the breeding environment, animal transhumance and breeding system.展开更多
Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding...Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus(PPRV)pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals,establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.展开更多
为实现小反刍兽疫病毒(Peste des petits ruminants virus,PPRV)的快速、准确、定量检测,以国内PPRV Clone 9疫苗株N基因为靶位点设计特异性引物及探针,建立了液滴式数字PCR(Droplet digital PCR,ddPCR)方法,并对其特异性、敏感性及重...为实现小反刍兽疫病毒(Peste des petits ruminants virus,PPRV)的快速、准确、定量检测,以国内PPRV Clone 9疫苗株N基因为靶位点设计特异性引物及探针,建立了液滴式数字PCR(Droplet digital PCR,ddPCR)方法,并对其特异性、敏感性及重复性进行评价。将所建立的ddPCR方法与实时荧光定量PCR(Fluorescence quantitative PCR,qPCR)方法的灵敏度进行比较分析并应用于PPRV(Clone 9株)核糖核酸标准物质的定量。结果显示,所建立的ddPCR方法特异性好,仅PPRV检测结果为阳性,其他羊源病毒类生物制品及毒种检测结果均为阴性;敏感性高,基因拷贝数检出限度为4.33拷贝/μL,比qPCR方法的灵敏度(57.3拷贝/μL)高10倍;重复性好,重复性试验的变异系数为1.8%;定量准确,具有一定资质的9家单位各组测量数据组间变异系数小于5%。试验建立的ddPCR方法能够有效地对PPRV核酸进行快速检测,为PPRV的诊断及绝对定量提供了有效方法。展开更多
基金This work was supported by project from MOA [2006-G57(3)B-Z1] Project from Yunnan Province (2008LA019)
文摘This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of which 2 samples were genetically close to the X7443 strain (Nigeria 75/1) of lineage I, and 3 samples close to the strain AY560591 (Sungri96) of linage IV with 96.6%、97.3%、97.6% and 98% nucleotide sequence homogeneity respectively, based on partial sequencing of the F gene from 5 samples and complete sequencing of the N/M/F/H genes from one sample. This study suggested that there are at least 2 origins of PPRV in China.
文摘This experiment was conducted to diagnose Peste des Petits Ruminants based on the eukaryotically-expressed PPRV nucleoprotein through an indirect ELISA. The full-length PPRV nucleoprotein gene was obtained from viral genome RNA by RT-PCR. The amplified fragments were cloned into baculovirus donor vectors pFastHTA of the Bac-to-Bac system. These recombinant plasmids, pFastHTA-PPRV-N, were transformed into DH10Bac host bacteria to obtain recombinant shuttle plasmids, pBacmid-PPRV-N. Recombinant baculovirus, Bacmid-PPRV-N, was generated for expression of the PPRV nucleoprotein by transfecting recombinant pBacmid-PPRV-N with Lipofectamine 2000 into Sf21 insect cells. The efficient expression of PPRV Nucleoprotein by baculovirus in Sf21 cells was verified by SDS-PAGE and Western blot. An indirect ELISA was developed using recombinant PPRV nucleoprotein as the coating antigen. 37 goat sera from an epidemic area in Tibet and 92 goat sera from a non-infected area in Qinghai Province were simultaneously detected by the indirect ELISA, developed here, and the international standard cELISA kit. The sensitivity and specificity of the indirect ELISA was 100% and 96.2% compared with the cELISA kit. The coincidence rate of the two methods was 96.9%. The results demonstrated that the indirect ELISA established in this study works well for diagnosis of PPR.
文摘Peste ties petits ruminants is a kind of acute eontagious disease infecting goats anti sheep. In this study, antibtly monitoring and tracking of healthy goat and sheep immunized by peste des petits ruminants vaccine in Changping District of Beijing City were conducted, aiming at providing a reference for the devel- opment of effective immunization procedures.
文摘Peste des petits ruminant (PPR) is a contagious disease of small ruminants caused by a virus that belongs to the genus Morbillivirus of the family Paramyxoviridae. This study aimed to determine the seroprevalence of PPR disease in sheep and goats and its associated risk factors in Kassala State, Eastern Sudan. Across sectional study was conducted during the period from 30th August to 25th November 2015. The study was carried out using a structured questionnaire survey and a total of 918 blood samples were collected from apparently healthy unvaccinated sheep and goats in different localities in State of Kassala. A total of 546 sheep and 372 goats were tested for specific antibodies to nucleoprotein (NP) by competitive enzyme linked immunosorbent assay (cELISA). The apparent overall prevalence of PPR antibodies in Kassala was 58.2% while the true prevalence was calculated to be 61.3%. The apparent prevalence in sheep and goats was 68.1% and 43.5% respectively. Univariate analysis showed that the risk factors had significant associations with a cELISA positive status: locality, species, age, breed, husbandry system, housing mode, animals movement (p = 0.000) and animals sharing pasture and water (p = 0.003), while sex and newly introduced animals were not significant risk factors (p = 0.771) (p = 0.050) respectively. Factors found that significantly associated (p < 0.05) with increased odds of being cELISA positive in multivariate analysis were localities, species, age and newly introduced animals. The prevalence differed between localities and was the highest in the River Atbara (84.0%) locality, whereas it was lowest in Delta North (29.0%). No significant difference was observed among the sexes. However, the prevalence differed in different age groups and was 52.25% in animals of less than six months old;49.3% were between seven months and two years old and 65.5% were above two years old. In different husbandry systems, the prevalence was 47.9%, 73.0% and 49.2% in intensive, open grazing and pastoral systems respectively. Housing type effects were also observed;the highest prevalence was in animals housed in metal fence (83.3%). The movement pattern showed significant effect, where the prevalence was the highest (81.3%) in animals that move inter-states/inter-localities. It is concluded that the disease is endemic in Kassala State, high prevalent in sheep and goats, posing a threat to animal exportation, and may have a serious economic influence. Owners and herders should compulsorily vaccinate their animals yearly and animals should be investigated periodically for implementation of crucial eradication program.
文摘The present study was carried out between April 2015 and January 2016 to estimate the sero-prevalence and identify the risk factors of the peste des petits ruminants (PPR) in Cameroon. A total of 269 herds randomly sampled across the country have been studied and 1622 samples of serum have been levied on the sheep and goat. The c-ELISA has been studied in order to detect the presence of antibodies in small ruminants like an indicator of exposition to PPRV. The results revealed the circulation of PPRV in the country with a total sero-prevalence of 39% [95%CI;37 - 41] and a sero-prevalence of 63.2% [95%CI;57.2 - 69.2] at the herd level. Sero-prevalence was variable in the ten regions ranging from 7% [95% CI;6.2 - 8.4] to 73% [95% CI;62 - 84] with the northern zone (Adamawa, North and Far-North) having 52.3% [95% CI;37 - 60] and southern zone (including the remaining seven regions) recording 29% [95% CI;11 - 57]. Similarly, it was higher in animals found in urban/peri-urban areas than in rural areas with prevalence ratio of 2.9 [95% CI 2.54 - 3.4;p < 0.001] <em>i.e. </em>3 times more, 1.6 [95% CI 1.36 - 1.90;p < 0.001] <em>i.e.</em> 1.6 times more, and 5.02 [95% CI 3.91 - 6.85;p < 0.001] <em>i.e.</em> 5 times more at national level, in the northern zone and in the southern area, respectively. Five risk factors have been identified: the breeding environment, introduction of new animals into the herds, gathering of animals for pasture and watering, wandering and transhumance. The breeding area appeared to be the most important risk factor associated with disease exposure. The control measures for the eradication of this disease must take into account the epidemiological situation, the breeding environment, animal transhumance and breeding system.
基金supported by the National Key Research and Development Program of China (2016YFD0500108 and 2016YFE0204100)the International Cooperation Project of CAAS Innovation Program (CAAS-GJHZ201700X)
文摘Peste des petits ruminants(PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus(PPRV)pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals,establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV.
