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Optimization and Scaling-Up of Peste des Petits Ruminants (PPR) Vaccine Production Using Roller Bottle Platform
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作者 Omer Algezoli Tagelden Nour +1 位作者 Muzdalifa Alamin Mohamed Abdalla 《Journal of Biosciences and Medicines》 2025年第1期378-393,共16页
Peste des Petits ruminants (PPR) is considered as one of the major constraints to the productivity of small ruminants in Sudan. Presently, control measures for PPR are primarily reliant on vaccination using an attenua... Peste des Petits ruminants (PPR) is considered as one of the major constraints to the productivity of small ruminants in Sudan. Presently, control measures for PPR are primarily reliant on vaccination using an attenuated PPR strain Nigeria 75/1 that has been produced in monolayers of Vero cells grown in static flasks. This study investigates the potential for scaling up PPR vaccine production using roller bottle technology, a more advanced method. A live, homologous vaccine against PPR in sheep and goats was successfully produced on a large scale in roller culture bottles, with DMEM supplemented with ten percent fetal bovine serum serving as the growth medium. The cells were infected with a multiplicity of infection of 0.01, and the vaccine was harvested when the cytopathic effect reached 80%. The vaccine was then freeze-dried to preserve its stability. A series of tests were conducted to ensure the safety and quality of the vaccine. Using PCR, the identity of vaccine was confirmed. It was found to be safe in both single and 100-times dose inoculations in sheep, with the produced batches showing a high titre of 6.4 ± 0.11 log10 TCID50/ml. All batches met the criteria of sterility, passing tests for bacteria, fungi, and mycoplasma. Furthermore, the vaccine proved effective in small ruminants, with antibodies persisting for over a year post-vaccination. The residual moisture content remained below 2.5%, and the vaccine successfully passed vacuum testing. Stability tests indicated that the vaccine has a shelf-life of at least one year when stored at temperatures of 2˚C - 8˚C and −20˚C. These results demonstrate the potential for applying roller bottle culture technology to PPR vaccine production, significantly streamlining the existing process and enhancing its efficiency. Further research is warranted to address the economic analyses of adopting roller bottle technology with existing PPR control program. 展开更多
关键词 Large-Scale Production Pest des petits ruminants Roller Bottle Technology VACCINATION Vero Cell
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Sero-Prevalence of Peste des Petits Ruminants Virus Antibodies in Sudanese Sheep and Goats before and after Vaccination
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作者 Omer Algezoli Selma Kamal +8 位作者 Yazeed Raouf Alamin Mozdalifa Hiba Ali Mohamed Aljameel Salih Sir Elkhatim Sulieman Ahmed Ibtesam Fadul Elsied Tageldin Nour Mohamed Abdalla 《Journal of Biosciences and Medicines》 2025年第1期21-40,共20页
Peste des petits ruminants virus (PPRV) antibodies were studied in Sudanese sheep and goats (n = 855) before and after vaccination with a locally produced Nigeria 75/1 vaccine using a commercial competitive ELISA (cEL... Peste des petits ruminants virus (PPRV) antibodies were studied in Sudanese sheep and goats (n = 855) before and after vaccination with a locally produced Nigeria 75/1 vaccine using a commercial competitive ELISA (cELISA) kit. Animals were kept healthy under field conditions, in four states: Blue Nile (n = 250), North Kordofan (n = 189), South Darfur (n = 225) and the Northern State (n = 191). Before vaccination, the overall sero-prevalence of PPRV antibodies was 54.6% (53.2% - 56%, 95% CI);high (64.8% - 76.4%, 95% CI) in Blue Nile State, medium (50.5% - 61.9%, 95% CI) in North Kordofan State and South Darfur State and low (28.6% - 35.2% 95%, CI) in Northern State. In high-risk areas (high sero-prevalence), Blue Nile (70.4%) and North Kordofan (57.7%), middle age groups (7 - 12 and 13 - 18 months) were identified as high-risk age. Middle age groups showed lower sero-prevalence than preceding (3 - 6 months) and subsequent (>18 months) age groups while the risk of exposure increased with age. Current and previous findings suggested a transmission pathway of PPRV involving the South Eastern border (Blue Nile) and neighbouring Central Sudan to North Kordofan. One month after vaccination 88.4% (343/388) of sero-negative animals were sero-converted suggesting the efficacy of the locally produced Nigeria 75/1 vaccine. Even if only individuals in the high-risk age group (7 - 18 months) were vaccinated, the overall population immunity (OPI) in high-risk areas (the Blue Nile and North Kordofan) would have surpassed the threshold of 70%, which is indicated for blocking PPRV transmission. However, lower vaccination coverage is expected in wider vaccination programmes. These findings primarily justified the targeting of PPR control in Sudan through the vaccination of high-risk age groups in high-risk areas. 展开更多
关键词 peste des petits ruminants (ppr) Vaccination Efficacy SEROPREVALENCE Herd Immunity High-Risk Area
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Establishment of goat infection model of the peste des petits ruminants virus isolated in China for vaccine efficacy evaluation
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作者 Xue Wang Hefeng Chen +14 位作者 Xianfeng Zhang Zhengshuang Wu Shuai Zhang Lei Shuai Lulu Wang Weijie Li Jinliang Wang Wenxing Liu Xijun Wang Zhiyuan Wen Jinying Ge Yuntao Guan Xijun He Weiye Chen Zhigao Bu 《Journal of Integrative Agriculture》 2025年第8期3199-3211,共13页
In 2013,peste des petits ruminants(PPR)re-emerged in China and spread to the majority of provinces across the country.The disease was effectively controlled through a vaccination campaign employing live attenuated vac... In 2013,peste des petits ruminants(PPR)re-emerged in China and spread to the majority of provinces across the country.The disease was effectively controlled through a vaccination campaign employing live attenuated vaccines,although sporadic cases still occurred.However,limited information is currently available regarding the peste des petits ruminants virus(PPRV)endemic in China.Here,a PPRV strain(HLJ/13)was isolated from a field sample in China using Vero cells expressing goat signalling lymphocyte activation molecule.Phylogenetic analysis indicated that HLJ/13 belonged to lineage IV.Subsequent intranasal and subcutaneous inoculation of goats with a dose of 2×10~6 TCID50of HLJ/13 resulted in the development of typical clinical symptoms of PPR,including pyrexia,ocular and nasal discharges,stomatitis,and diarrhea.All infected goats succumbed to the disease by day 8.To gain further insight,viral loading,pathological examination and immunohistochemical analyses were conducted,elucidating the main targets of HLJ/13 as the respiratory system,digestive tract and lymphoid organs.Employing the goat infection model established above,the goat poxvirus-vectored PPR vaccine,which was previously developed and could be used as DIVA(differentiating infected from vaccinated animals)vaccine,provided complete protection against the challenge of HLJ/13.It is important to note that this study represents the first comprehensive report delineating the biology and pathogenicity characterization,and infection model of PPRV isolated in China. 展开更多
关键词 peste des petits ruminants virus Chinese isolate goat infection model vaccine efficacy evaluation
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Evaluation of the Immunization Effect of a Peste des Petits Ruminants-Goatpox Combined Live Vaccine in Large-Scale Sheep Farms in Northern Shandong
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作者 Weiqin MENG Jinlong CHEN +5 位作者 Lu GUO Jianjun WANG Qiang FU Likun CHENG Shijin GUO Jinliang WANG 《Agricultural Biotechnology》 2025年第6期25-27,共3页
[Objectives]This study was conducted to enhance the prevention and control of peste des petits ruminants(PPR)and goatpox(GTP).[Methods]Two experimental sheep farms in Northern Shandong were selected to conduct a compa... [Objectives]This study was conducted to enhance the prevention and control of peste des petits ruminants(PPR)and goatpox(GTP).[Methods]Two experimental sheep farms in Northern Shandong were selected to conduct a comparative experiment between separate vaccinations for PPR and GTP and immunization with a combined live vaccine for both diseases.Antibody levels were measured to assess immunization effect on days 7,14,21 and 28 after vaccination.[Results]The qualified rates of group immune antibodies in both the experimental and control groups exceeded 75%,achieving the goal of preventing both PPR and GTP through a single immunization.[Conclusions]This study provides clinical application references for the prevention of PPR and GTP in the local area. 展开更多
关键词 peste des petits ruminants Goatpox Combined live vaccine
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Reverse Genetics for Peste des Petits Ruminants Virus: Current Status and Lessons to Learn from Other Non-segmented Negative-Sense RNA Viruses 被引量:4
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作者 Alfred Niyokwishimira Yongxi Dou +2 位作者 Bang Qian Prajapati Meera Zhidong Zhang 《Virologica Sinica》 SCIE CAS CSCD 2018年第6期472-483,共12页
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
关键词 peste des petits ruminants(ppr) pprV Reverse genetics Non-segmented negative-sense RNA virus
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Monitoring and Tracking on Immune Antibody of Sheep Peste des Petits Ruminants 被引量:1
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作者 Lv Yanqiu Huang Dongfeng +8 位作者 Wang Meili Wang Jinxia Wang Yuewei Qiu Hailian Gao Xiaobo Zhang Yichi Kang Xiaojie Shan Lihua Xue Yong 《Animal Husbandry and Feed Science》 CAS 2017年第2期96-97,共2页
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
关键词 MONITORING Immune antibody Sheep peste des petits ruminants
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Seroprevalence and Associated Risk Factors of Peste des Petits Ruminants among Sheep and Goats in Kassala State, Sudan
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作者 Fatima A. Saeed Sana A. Abdel-Aziz Mohammed M. Gumaa 《Open Journal of Animal Sciences》 2018年第4期381-395,共15页
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
关键词 peste des petits ruminants SERO-PREVALENCE Risk Factors Kassala STATE SUDAN
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Sero-Prevalence and Risk Factors of Diffusion of Peste des Petits Ruminants in Cameroon
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作者 Severin Loul Abel Wade Alexandre Michel Njan Nlôga 《Open Journal of Veterinary Medicine》 2020年第7期103-115,共13页
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
关键词 Small ruminants SERO-PREVALENCE Risk Factors peste des petits ruminants Cameroon
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Establishment of High-sensitivity Rapid Fluorescence Quantitative Detection Method for Antibody against Peste des Petits Ruminants Virus
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作者 Zhao LIU Bo LIU +3 位作者 Zhida LIN Hang SUN Yu SUN Xiaohui SONG 《Agricultural Biotechnology》 2024年第5期22-27,共6页
[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
关键词 peste des petits ruminants N protein NH fusion protein Soluble expression and purification Rapid quantitative detection
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Establishment of Double-antigen Sandwich Time-resolved Fluorescence Immunoassay for Detection of Pest des Petits Ruminants Virus
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作者 Binglei CAO Zhongyuan GE +3 位作者 Qi YANG Hang SUN Yu SUN Xiaohui SONG 《Agricultural Biotechnology》 2024年第4期21-27,共7页
[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
关键词 peste des petits ruminants N active protein NH fusion protein Soluble expression and purification Time-resolved fluorescence immunoassay
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基于MS2噬菌体的小反刍兽疫装甲RNA质控品的研制
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作者 刘丹 李慧彤 +8 位作者 张博源 高建帅 蒋卉 范学政 张广智 郜卫华 丁家波 熊涛 沈青春 《中国兽医杂志》 北大核心 2025年第5期43-51,共9页
为了研制安全稳定的小反刍兽疫病毒(PPRV)核酸检测质控品,本试验将PPRV检测靶标基因N、MS2噬菌体衣壳蛋白(CP)和成熟酶蛋白A基因序列插入p ET28a构建重组质粒p ET-CPA-PPR,经表达纯化、去除残余核酸和实时荧光定量PCR鉴定后制备成PPRV装... 为了研制安全稳定的小反刍兽疫病毒(PPRV)核酸检测质控品,本试验将PPRV检测靶标基因N、MS2噬菌体衣壳蛋白(CP)和成熟酶蛋白A基因序列插入p ET28a构建重组质粒p ET-CPA-PPR,经表达纯化、去除残余核酸和实时荧光定量PCR鉴定后制备成PPRV装甲RNA质控品,并对其进行电子显微镜观察、定量、耐核酸酶检验、均匀性和稳定性检验,以评估其作为标准物质的可能性。结果显示,p ET-CPA-PPR经诱导表达和纯化后形成直径为23~28 nm、大小均一、已包封PPRV检测靶标基因N的装甲RNA颗粒,浓度为2.24×10^(9)copies/μL,可以有效抵抗核酸酶降解作用,均匀性良好且在(37±1)℃条件下可稳定保存至少30 d。结果表明,基于MS2噬菌体制备的PPRV装甲RNA拷贝数高,均匀性和稳定性良好,能够为PPRV核酸检测提供安全可靠的标准化参考物质。 展开更多
关键词 小反刍兽疫(ppr) MS2噬菌体 装甲RNA 病毒样颗粒
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2022—2024年湖南省常宁市羊小反刍兽疫抗体监测结果分析
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作者 詹逸舒 谭运华 《现代畜牧兽医》 2025年第7期59-63,共5页
研究旨在评估常宁市羊小反刍兽疫(PPR)的免疫成效,进一步做好羊PPR的防控工作,统计并分析了常宁市2022—2024年羊PPR的免疫抗体监测结果。结果显示:2022—2024年常宁市羊PPR抗体样品总体合格率为86.87%(1 905/2 193),养殖场总体合格率为... 研究旨在评估常宁市羊小反刍兽疫(PPR)的免疫成效,进一步做好羊PPR的防控工作,统计并分析了常宁市2022—2024年羊PPR的免疫抗体监测结果。结果显示:2022—2024年常宁市羊PPR抗体样品总体合格率为86.87%(1 905/2 193),养殖场总体合格率为84.62%(99/117),且逐年稳步提高;规模场羊PPR抗体样品合格率为90.66%(1 252/1 381),场合格率为88.16%(67/76);散养户的羊PPR抗体样品合格率80.42%(653/812),户合格率为78.05%(32/41);春季羊PPR抗体样品合格率为84.48%(882/1 044),场合格率为80.00%(44/55);秋季羊PPR抗体样品合格率89.03%(1 023/1 149),户合格率为88.71%(55/62);公羊PPR免疫抗体的个体合格率为89.10%(1 439/1 615),母羊合格率为80.62%(466/578);不同乡镇羊PPR免疫抗体合格率最高为96.65%,最低为76.26%。规模场样品和场PPR免疫抗体合格率均高于散养户(P<0.05),秋季样品和场(户) PPR免疫抗体合格率均高于春季(P<0.05),公羊样品PPR免疫抗体合格率均高于母羊(P<0.05)。研究表明,常宁市2022—2024年PPR抗体免疫合格率超过了农业农村部规定标准(≥70%),免疫抗体水平较高且逐年稳步提高,免疫成效好,达到了预期的免疫效果。 展开更多
关键词 小反刍兽疫 抗体监测 常宁市
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表达小反刍兽疫H蛋白的重组山羊痘病毒疫苗 被引量:7
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作者 陈伟业 曲林茂 +5 位作者 胡森 胡倩倩 张倩 支海兵 黄克和 步志高 《生物工程学报》 CAS CSCD 北大核心 2009年第4期496-502,共7页
本研究旨在构建表达小反刍兽疫H蛋白的重组山羊痘病毒,并评价其免疫效力。利用筛选基因gpt和eGFP并结合空斑技术,纯化筛选了重组小反刍兽疫H基因的重组山羊痘病毒(rGPV-PPRV-H),经过PCR鉴定重组病毒已纯化。免疫荧光和蛋白印迹都表明重... 本研究旨在构建表达小反刍兽疫H蛋白的重组山羊痘病毒,并评价其免疫效力。利用筛选基因gpt和eGFP并结合空斑技术,纯化筛选了重组小反刍兽疫H基因的重组山羊痘病毒(rGPV-PPRV-H),经过PCR鉴定重组病毒已纯化。免疫荧光和蛋白印迹都表明重组病毒能够感染绵羊羔羊睾丸细胞表达小反刍兽疫H蛋白。以2×106PFU的rGPV-PPRV-H皮内注射免疫山羊6只,并于首次免疫后28d以相同剂量进行二次免疫。免疫后采血分离血清进行病毒中和试验,结果表明,一次免疫后21d,山羊痘病毒中和抗体效价依次为40、80、≥80、≥80、40、≥80,和小反刍兽疫病毒中和抗体效价全部转阳依次为80、80、80、80、40、40、10;二次免疫后14d,山羊痘病毒中和抗体效价抗体全部大于或等于80,小反刍兽疫病毒中和抗体效价依次为≥80、80、≥80、80、80、40,应该具有对山羊痘病毒和小反刍兽疫病毒强毒攻击的完全免疫保护作用。本研究为小反刍兽疫重组山羊痘疫苗的产业化提供了参考。 展开更多
关键词 重组疫苗 小反刍兽疫 山羊痘病毒载体 中和抗体
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小反刍兽疫病毒N蛋白主要抗原区域的原核表达及间接ELISA检测方法的建立 被引量:10
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作者 邱文英 李刚 +1 位作者 田康乐 黄华欣 《中国兽医科学》 CAS CSCD 北大核心 2012年第5期483-487,共5页
为了建立可特异性地检测血清中抗小反刍兽疫病毒(PRRV)抗体的方法,对具有小反刍兽疫病毒特异性的N蛋白第Ⅳ区域进行扩增,并克隆至原核表达载体pET-32a(+)中,用于小反刍兽疫病毒N亚单位重组蛋白的表达。用纯化的重组蛋白作为检测抗原,建... 为了建立可特异性地检测血清中抗小反刍兽疫病毒(PRRV)抗体的方法,对具有小反刍兽疫病毒特异性的N蛋白第Ⅳ区域进行扩增,并克隆至原核表达载体pET-32a(+)中,用于小反刍兽疫病毒N亚单位重组蛋白的表达。用纯化的重组蛋白作为检测抗原,建立了检测血清中抗小反刍兽疫病毒抗体的间接ELISA方法,并对该间接ELISA的抗原包被量、血清稀释倍数、血清孵育时间等反应条件进行了优化。Western-blot分析结果表明,重组蛋白具有良好的反应原性。所建立的间接ELISA方法可以鉴别血清中的小反刍兽疫病毒抗体和牛瘟病毒抗体,具有良好的特异性和灵敏性,与标准竞争ELISA试剂盒的符合率达98.4%。证实,本研究建立的检测血清中抗PPRV抗体的间接ELISA方法具有良好的特异性和灵敏性。 展开更多
关键词 小反刍兽疫 N蛋白 间接酶联免疫吸附试验
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小反刍兽疫病毒的研究进展 被引量:23
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作者 李林杰 谢军 +2 位作者 徐文凯 马晓霞 柏家林 《黑龙江畜牧兽医》 CAS 北大核心 2018年第1期66-70,252,共6页
小反刍兽疫(peste des petits ruminants,PPR)是由一种副黏病毒科麻疹病毒属病毒引起的主要感染山羊、绵羊的急性高治病性传染病,但牛感染后临床症状不明显。世界动物卫生组织(OIE)将其列为A类疫病,感染率和死亡率高达90%。由于绵羊和... 小反刍兽疫(peste des petits ruminants,PPR)是由一种副黏病毒科麻疹病毒属病毒引起的主要感染山羊、绵羊的急性高治病性传染病,但牛感染后临床症状不明显。世界动物卫生组织(OIE)将其列为A类疫病,感染率和死亡率高达90%。由于绵羊和山羊是非洲和西亚贫困人民的重要生产资料,PPR对食品安全和摆脱贫困构成巨大威胁。小反刍兽疫病毒(PPRV)和牛瘟病毒(RPV)与麻疹病毒(MV)联系紧密。通过大规模接种疫苗牛瘟已被彻底消除。虽然可以利用弱毒疫苗免疫机体预防PPRV感染,但由于其在亚热带气候中的热不稳定性、使用时所需剂量的不确定性及接种范围的不足导致该病仍未得到有效控制。另外已有证据表明,在疫苗与不同毒株之间存在很少的交叉中和,使得目前流通的PPRV疫苗的保护功效被质疑。文章简要介绍了目前全世界对PPRV的高关注度及PPRV的生物学特性、发病机理等。 展开更多
关键词 小反刍兽疫(ppr) 小反刍兽疫病毒(pprV) 蛋白 结构 病毒复制
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全球小反刍兽疫流行趋势 被引量:17
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作者 朱迪国 宋建德 +1 位作者 袁丽萍 魏荣 《中国动物检疫》 CAS 2014年第6期14-16,共3页
2013年以来,全球共43个国家报告发生小反刍兽疫(PPR)。其中,疫情仍然集中在亚洲和非洲。本文介绍了全球小反刍兽疫的发生情况,尤其是我国周边国家的疫情情况,分析了全球小反刍兽疫的扩散趋势。
关键词 小反刍兽疫 流行 趋势
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小反刍兽疫病毒的分子生物学特性及其在全球的流行 被引量:21
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作者 罗静 何宏轩 《河北师范大学学报(自然科学版)》 CAS 北大核心 2009年第4期543-550,共8页
小反刍兽疫由副粘病毒科麻疹病毒属小反刍兽疫病毒引起,经直接或间接接触传染,主要感染山羊、绵羊等小反刍动物,严重影响畜牧业,造成巨大经济损失.1942年首次发现于西非科特迪瓦,逐渐扩散至非洲及亚洲大部,涉及四十余国家.2007年7月首... 小反刍兽疫由副粘病毒科麻疹病毒属小反刍兽疫病毒引起,经直接或间接接触传染,主要感染山羊、绵羊等小反刍动物,严重影响畜牧业,造成巨大经济损失.1942年首次发现于西非科特迪瓦,逐渐扩散至非洲及亚洲大部,涉及四十余国家.2007年7月首次传入中国西藏阿里地区,并于2008年6月再度爆发,且呈由边境逐渐向内传播的趋势.该地区有多种珍贵濒危野生反刍动物,是小反刍兽疫病毒的潜在宿主,它们随季节迁徙及夏季产仔的习性更增加其受小反刍兽疫感染并传播小反刍兽疫的可能.因此,对小反刍兽疫病毒的生物学特征、流行特点和防控措施进行综述,必将有利于正确评估小反刍兽疫对野生有蹄类动物的威胁并采取合适的措施预防与控制小反刍兽疫疫情. 展开更多
关键词 小反刍兽疫 分子生物学特性 流行 野生有蹄类动物
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小反刍兽疫病毒样颗粒的构建及定量分析 被引量:1
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作者 邓俊花 林祥梅 +2 位作者 张永宁 王彩霞 吴绍强 《中国兽医科学》 CAS CSCD 北大核心 2014年第10期997-1002,共6页
为研制一种无生物传染性且极易保存稳定的小反刍兽疫病毒(PPRV)RNA阳性质控品,以pTrcHis-MS2质粒为模板,借助点突变技术在噬菌体MS2编码的外壳蛋白第15位与第16位氨基酸基因之间插入组氨酸蛋白标签的相应序列,构建了pTrcMS载体。将pGEM-... 为研制一种无生物传染性且极易保存稳定的小反刍兽疫病毒(PPRV)RNA阳性质控品,以pTrcHis-MS2质粒为模板,借助点突变技术在噬菌体MS2编码的外壳蛋白第15位与第16位氨基酸基因之间插入组氨酸蛋白标签的相应序列,构建了pTrcMS载体。将pGEM-T-N382质粒与pTrcMS质粒分别进行KpnⅠ+HindⅢ双酶切、连接从而构建了pTrcMS-PPRV重组质粒。将pTrcMS-PPRV重组菌进行原核表达,借助偶联组氨酸蛋白标签的磁珠捕获技术获得纯化的小反刍兽疫病毒样颗粒,对它进行特性鉴定并定量检测。结果显示,pTrcMS-PPRV病毒样物质纯度高;电镜下为不规则的、直径约为26nm的多边形;耐RNA酶A,稳定;经荧光RT-PCR定量检测,此病毒样物质的溶液浓度相当于108.56 PFU/mL。本研究将为小反刍兽疫病毒分子生物学的检测提供阳性质控物质。 展开更多
关键词 小反刍兽疫 病毒样颗粒 组氨酸蛋白标签 纯化 定量分析
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小反刍兽疫活疫苗的安全性评价试验 被引量:5
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作者 薛青红 印春生 +1 位作者 李宁 支海兵 《中国兽药杂志》 2010年第9期4-6,共3页
2007年小反刍兽疫(PPR)在我国西藏首次暴发,在西藏和新疆部分地区使用PPR Nigeria 75/1疫苗株制造的疫苗进行免疫接种。为明确疫苗的安全性,中国兽医药品监察所国家牛瘟参考实验室对其安全性能进行了系统评价。健康易感山羊、绵羊及怀... 2007年小反刍兽疫(PPR)在我国西藏首次暴发,在西藏和新疆部分地区使用PPR Nigeria 75/1疫苗株制造的疫苗进行免疫接种。为明确疫苗的安全性,中国兽医药品监察所国家牛瘟参考实验室对其安全性能进行了系统评价。健康易感山羊、绵羊及怀孕山羊、怀孕绵羊按不同剂量接种疫苗后,均未观察到异常临床反应;怀孕母羊所产羔羊数量与对照组无明显差异。疫苗对小白鼠、豚鼠的非特异性安全试验表明,所有接种动物均健活。结果表明该疫苗安全性良好,可在田间大规模使用。 展开更多
关键词 小反刍兽疫 疫苗 安全
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小反刍兽疫免疫荧光诊断技术研究 被引量:1
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作者 蒋梅 赵文华 杨仕标 《黑龙江畜牧兽医》 CAS 北大核心 2012年第4期102-105,174,共5页
为了建立小反刍兽疫(PPR)荧光抗体诊断方法,研究选用小反刍兽疫病毒(PPRV)Nigeria75/1减毒疫苗株制备抗原,免疫试验山羊,制备小反刍兽疫高免血清,用硫酸铵盐析法粗提免疫球蛋白(IgG),再应用提取的IgG制备异硫氰酸荧光素标记抗体。结果表... 为了建立小反刍兽疫(PPR)荧光抗体诊断方法,研究选用小反刍兽疫病毒(PPRV)Nigeria75/1减毒疫苗株制备抗原,免疫试验山羊,制备小反刍兽疫高免血清,用硫酸铵盐析法粗提免疫球蛋白(IgG),再应用提取的IgG制备异硫氰酸荧光素标记抗体。结果表明:所制备的荧光抗体对试验用Vero细胞组织抗原和犬瘟热病毒(CDV)抗原无反应性,对小反刍兽疫病毒感染细胞的检出敏感性为1×106稀释度。 展开更多
关键词 小反刍兽疫(ppr) 诊断方法 荧光抗体
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