[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PP...[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.展开更多
[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were ...[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E.coli.Furthermore,based on the purified soluble N protein and NH fusion protein,a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established.[Results]The method could quickly and quantitatively detect PPR V antibody in sheep serum,with high sensitivity and specificity,without any cross reaction to other related sheep pathogens.The intra-batch and inter-batch coefficients of variation were less than 10%and 15%,respectively,and the method had good repeatability.Through detection on 292 clinical serum samples,it was compared with the French IDVET competitive ELISA kit,and the coincidence rate of the two methods reached 93.84%.Compared with the serum neutralization test,the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum(provided by the WOAH Brucellosis Reference Laboratory of France).[Conclusions]This method can realize rapid quantitative detection of PPR V antibody on site,and has high practical value and popularization value.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
[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants （PPR）. [ Method] Soluble N protein with high expression was obtained from Escherichia c...[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants （PPR）. [ Method] Soluble N protein with high expression was obtained from Escherichia coli expression system through codon optimization and optimization of expression conditions, and indirect ELISA detection method based on N protein was further established. [ Result] The assay had no cross reaction with other sheep pathogens. The intra- and inter-batch variation coefficients were less than 9%, indicating the method had good repeatability. Furthermore, totally 480 clinical serum samples were detec- ted by the assay, and the agreement rate with commercial ELISA kit （IDVET） was 98.33%. [ Conclusion] The study laid a foundation for further development of mature PPRV antibody detection kits.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
Dashilan et Liulichang,situés au centre-ville de Beijing,sont des quartiers historiques et culturels renommés.Un musée discret se niche dans le dédale de petites maisons aux murs gris:le Mus&...Dashilan et Liulichang,situés au centre-ville de Beijing,sont des quartiers historiques et culturels renommés.Un musée discret se niche dans le dédale de petites maisons aux murs gris:le Musée de la cour n°93.Depuis 2014,il favorise la diffusion du patrimoine culturel immatériel et des arts populaires.展开更多
基金Supported by National Key R&D Program for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39)+2 种基金Key Research and Development Program of Shandong Province(Major Science and Technology Innovation Project)(2021CXGC011306)Scientific Research Project of General Administration of Customs(2024HK033)Scientific Research Project of Jinan Customs(2023JK005).
文摘[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.
基金Supported by The National Project for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39).
文摘[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E.coli.Furthermore,based on the purified soluble N protein and NH fusion protein,a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established.[Results]The method could quickly and quantitatively detect PPR V antibody in sheep serum,with high sensitivity and specificity,without any cross reaction to other related sheep pathogens.The intra-batch and inter-batch coefficients of variation were less than 10%and 15%,respectively,and the method had good repeatability.Through detection on 292 clinical serum samples,it was compared with the French IDVET competitive ELISA kit,and the coincidence rate of the two methods reached 93.84%.Compared with the serum neutralization test,the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum(provided by the WOAH Brucellosis Reference Laboratory of France).[Conclusions]This method can realize rapid quantitative detection of PPR V antibody on site,and has high practical value and popularization value.
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.
基金Supported by New Diagnosis and Detection Technology Research for Major Animal Diseases in Cattle and Sheep(No.2016YFD0500901)
文摘[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants （PPR）. [ Method] Soluble N protein with high expression was obtained from Escherichia coli expression system through codon optimization and optimization of expression conditions, and indirect ELISA detection method based on N protein was further established. [ Result] The assay had no cross reaction with other sheep pathogens. The intra- and inter-batch variation coefficients were less than 9%, indicating the method had good repeatability. Furthermore, totally 480 clinical serum samples were detec- ted by the assay, and the agreement rate with commercial ELISA kit （IDVET） was 98.33%. [ Conclusion] The study laid a foundation for further development of mature PPRV antibody detection kits.
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.
文摘Dashilan et Liulichang,situés au centre-ville de Beijing,sont des quartiers historiques et culturels renommés.Un musée discret se niche dans le dédale de petites maisons aux murs gris:le Musée de la cour n°93.Depuis 2014,il favorise la diffusion du patrimoine culturel immatériel et des arts populaires.
