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pcv-2和pcv-3 Cap蛋白在昆虫细胞-杆状病毒系统中的表达和鉴定

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中国兽医杂志 2023年第6期59卷 55-59页

作者：董宇璠金宏凯孙莉费东亮马跃宇马鸣潇李明锦州医科大学畜牧兽医学院辽宁锦州121000 锦州医科大学生命科学研究院辽宁锦州121000 锦州医科大学实验动物中心辽宁锦州121000

为了实现猪圆环病毒2型(pcv-2)和3型(pcv-3)Cap蛋白在昆虫细胞-杆状病毒系统中表达,本试验利用刚性连接肽连接pcv-2和pcv-3 Cap基因,构建杆状病毒转移载体pFastBac-pcv-2-3-Cap,双酶切验证正确后,将其转化至大肠杆菌DH10Bac感受态细胞,... 详细信息

为了实现猪圆环病毒2型(pcv-2)和3型(pcv-3)Cap蛋白在昆虫细胞-杆状病毒系统中表达,本试验利用刚性连接肽连接pcv-2和pcv-3 Cap基因,构建杆状病毒转移载体pFastBac-pcv-2-3-Cap,双酶切验证正确后,将其转化至大肠杆菌DH10Bac感受态细胞,经蓝白斑筛选和PCR鉴定,获得重组杆粒。将重组杆粒转染SF9细胞(昆虫卵巢细胞),盲传3代,获得重组病毒。将重组病毒接种SF9细胞96 h后,通过细胞免疫荧光检测、Western blot和电镜观察对其进行验证,并利用噬斑法检测其效价。结果显示:成功构建重组质粒pFastBac-pcv-2-3-Cap,获得了重组杆粒rAcBacmid-pcv-2-3-Cap,并制备出重组杆状病毒rvAc-pcv-2-3-Cap。重组病毒感染SF9细胞后,目的蛋白成功表达,并可以形成病毒样颗粒,病毒效价为3.41×10^(6) PFU/mL。本试验成功在昆虫细胞-杆状病毒系统中共表达了pcv-2-3-Cap,为防治pcv-2和pcv-3混合感染的二联疫苗研究提供了数据支持。

关键词： pcv-2 pcv-3 重组质粒杆状病毒真核表达

Virus Location and Lymphocyte Apoptosis in Lymph Nodes of Piglets Infected with pcv-2

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Agricultural Sciences in China 2008年第4期7卷 507-512页

作者： SHI Lū-juan HAN Hui-li ZHANG Shu-xia College of Veterinary Medicine Nanjing Agricultural University Nanjing 210095 P.R.China

The present study has been performed to understand the location of the virus, type of apoptotic cells, and their relation to lymph nodes of piglets infected with porcine circovirus type Ⅱ （pcv-2）. Nine 32-day-old conventional piglets free of infection with pcv-2 were used, and distributed into three groups： control group （n = 3）, piglets inoculated with pcv-2 alone （pcv-2, n = 3）, and pcv-2 inoculated and KLH immunostimulated group （pcv-2 ＋ KLH, n = 3）. Superficial inguinal lymph nodes from all piglets were collected for histological examination after 32 days postinoculation, and immunohistochemistry for pcv-2 detection. Location of apoptotic cells was detected with TdT-mediated dUTP nick end labeling （TUNEL） and cell cycle, and the apoptotic rates were measured by flow cytometry. The characteristic histopathological lesions of the piglets in pcv-2 and pcv-2 ＋ KLH were lymphocyte depletions in the cortex and paracortex of the lymph nodes, epithelioid-like macrophage infiltration, and intracytoplasmic inclusion bodies presented in epithelioid-like macrophages. pcv-2 was mainly found in epithelioid-like macrophages by immunohistochemistry. In the lymph nodes, lymphocytes presented higher apoptotic rates in the cortex by TUNEL, special B-cell areas, and similar apoptotic cells were found in this compartment in the control. The apoptotic rates of the lymph nodes were 0.41, 3.34, and 4.88% in the control, pcv-2, and pcv-2 ＋ KLH groups by flow cytometry, respectively. The apoptotic rates of lymph nodes for pcv-2 and pcv-2 ＋ KLH piglets were significantly higher than those for the control group （P〈0.05 and P〈0.01）. The proliferation index （PI） was 0.17_＋0.01, 0.12_＋0.01 and 0.12_＋0.04 in the control, pcv-2, and pcv-2 ＋ KLH group, the PI of the control group was higher than that of the other groups, but without the statistical difference. pcv-2 can induce lymphocyte depletion in lymph nodes of piglets by blocking cell proliferation and promoting apoptosis. This is one of th

关键词： apoptosis virus located lymph node pcv-2 piglets

某猪场致生长育肥猪腹泻的猪圆环病毒2型(pcv-2)病原学检测与病理组织学观察

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家畜生态学报 2023年第10期44卷 89-91页

作者：张永康杨秀进宁夏职业技术学院宁夏银川750021 北京科牧丰生物制药有限公司北京102600

某猪场80日龄左右的生长育肥猪群出现顽固性腹泻,腹泻粪便的形状复杂,为了查明病原,该试验采集病猪病料,提取病毒DNA和RT-PCR检测,并进行病理变化分析和病理组织学观察。结果表明:实时荧光定量PCR检测显示猪圆环病毒2型(pcv-2)强阳性,... 详细信息

某猪场80日龄左右的生长育肥猪群出现顽固性腹泻,腹泻粪便的形状复杂,为了查明病原,该试验采集病猪病料,提取病毒DNA和RT-PCR检测,并进行病理变化分析和病理组织学观察。结果表明:实时荧光定量PCR检测显示猪圆环病毒2型(pcv-2)强阳性,剖检发现脾脏有轻微梗死、淋巴结萎缩,病理组织学观察发现回肠肠粘膜上皮细胞显著增生,由单层变为多层,淋巴结和脾脏的淋巴细胞数量显著减少。试验说明pcv2是引起该猪场育肥猪腹泻的主要病原。

关键词：生长育肥猪腹泻 pcv-2 病原学检测病理组织学观察

Development of a drtqPCR Assay for Differentiation of pcv-2a and 2b in Shanghai Area

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Open Journal of Veterinary Medicine 2014年第12期4卷 286-293页

作者： Jian Liu Feifei Ge Houbing Ju Dequan Yang Xin Li Jian Wang Peihong Liu Jinping Zhou Shanghai Animal Disease Control Center Shanghai China

A dual real-time quantitative polymerase chain reaction assay (drtqPCR) was established to detect and differentiate between Porcine circovirus-2a (pcv-2a) and Porcine circovirus-2b (pcv-2b). Genotype-specific primer sets and probes were designed by using sequence data published for different pcv-2 strains. Specificity and sensitivity of the drtqPCR were examined by using pcv-2 isolates with known genotype. Among 367 tissue samples, 44.69% (164/367) were pcv-2 positive. From 164 pcv-2 positive samples, 10.98% (18/164), 92.56% (137/164), and 3.31% (9/164) were positive for pcv-2a, pcv-2b, and both genotypes, respectively. These results suggest that the dif-ferential drtqPCR can be used to detect pcv-2 and to differentiate the 2 genotypes from field sam-ples. The pcv-2 infection is quite common in swine of Shanghai area. Furthermore, the pcv-2b infective ratio is far higher than pcv-2a, and pcv-2a/2b mixed infections are also observed but at a lower prevalence in Shanghai area.

关键词： Genotype Differentiation drtqPCR pcv-2

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pcv-2对自然感染猪群生产性能的影响

黑龙江畜牧兽医（下半月） 2017年第12期 119-121页

作者：李瑞香吴奇强付小伟渭南市临渭区动物疾病控制中心陕西渭南714000 渭南市动物疾病控制中心陕西渭南714000

猪圆环病毒在猪群中感染已较为普遍,但仅有少数猪表现出明显的临床症状,大部分为隐性感染,常常表现为渐进性消瘦、皮肤苍白、腹泻、母猪繁殖障碍等症状,血清流行病学调查结果表明：猪圆环病毒2型（pcv-2）感染呈世界性分布,感染后常造... 详细信息

猪圆环病毒在猪群中感染已较为普遍,但仅有少数猪表现出明显的临床症状,大部分为隐性感染,常常表现为渐进性消瘦、皮肤苍白、腹泻、母猪繁殖障碍等症状,血清流行病学调查结果表明：猪圆环病毒2型（pcv-2）感染呈世界性分布,感染后常造成机体免疫系统功能下降[1-3],从而可促使其他病原微生物的并发或者继发感染,使猪群发病率和病死率显著升高,严重制约着养猪业的发展。

关键词：自然感染 pcv-2 猪群生产性能猪圆环病毒2型母猪繁殖障碍血清流行病学免疫系统功能

Investigation on the co-infections of Toxoplasma gondii with PRRSV,CSFV or pcv-2 in swine in part of China

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Investigation on the co-infections of Toxoplasma gondii with...

表达PRRSV ORF5和pcv2 ORF2基因的重组鸡痘病毒的构建及鉴定

中国毒理学会兽医毒理学委员会与中国畜牧兽医学会兽医食品卫生学分会联合学术研讨会暨中国毒理学会兽医毒理学委员会第5次全国会员代表大会

作者： Shuaiwang Meng Zhang Xinchao Liu Tao Liu HanchunYang ShishanYuan Guangwei Zhao Hassan I.A Ruofeng Yan Xiaokai Song Lixin Xu Xiangrui Li College of Veterinary Medicine Nanjing Agricultural University

【Objective】The objective of the present investigation was to estimate the prevalence of Toxoplasma gondii infection and co-infection with porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV)and porcine circovirus type 2(pcv-2)in pigs in China.【Method】A total of 372 tissues or serum samples collected from pigs distributed in 9provinces/municipalities of China during the period from February 2011 to November 2012 were assayed for *** antigens and antibodies using enzyme linked immunosorbent assay(ELISA)technique,while the PCR was designed for the detection of the PRRSV,CSFV and pcv-2,respectively.【Result】The total seropositive rate of *** was found in 34(9.14%)out of 372 samples.A total of186 samples were positive for PRSSV and the positive rate was 50.00%.The positive rates of CSFV and pcv-2 were 37.10%(138/372)and 3.23%(12/372),*** the 34 *** sero-positive samples,8 samples were infected with *** alone,while the remaining 26 samples were simultaneously infected with *** and one of the PRSSV,CSFV and pcv-2,or the combination of 3 *** co-infection rate of *** with PRSSV,*** with PRSSV and CSFV,*** with PRSSV and pcv-2,*** with CSFV and pcv-2,*** with PRSSV,CSFV and pcv-2 was 1.61%(6/372),4.03%(15/372),0.27%(1/372),0.27%(1/372)and 0.80%(3/372),respectively.【Conclusions】The results of the present survey revealed that PRRSV and CSFV were the common pathogens co-existing with porcine toxoplasmosis in China,and both of them could increase the chances of *** infection in *** results of the present survey indicated that *** infection in pigs was as high as 9.14%in China,and the high infection of *** in pigs might be a risk to the safety of animal origin food and public *** and CSFV were the common pathogens co-existing with porcine toxoplasmosis in China,and both of them could increase the chances of *** infection in *** is th

关键词： Toxoplasma gondii co-infection PRRSV CSFV pcv-2 pig

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中国预防兽医学报 2007年第6期29卷 405-409页

作者：沈国顺金宁一庄天中韩贞珍李阳张金双吉林大学畜牧兽医学院吉林长春130062 军事医学科学院军事兽医研究所吉林长春130062

分别将猪圆环病毒2型的ORF2基因和猪繁殖与呼吸综合征病毒的ORF5基因,插入到鸡痘病毒转移载体pUTA2-16-LacZ单一启动子和复合启动子下游,构建重组鸡痘病毒转移质粒pUTAL-ORF5-ORF2。将该重组质粒与鸡痘病毒282-E4株共转染鸡胚成纤维细胞... 详细信息

分别将猪圆环病毒2型的ORF2基因和猪繁殖与呼吸综合征病毒的ORF5基因,插入到鸡痘病毒转移载体pUTA2-16-LacZ单一启动子和复合启动子下游,构建重组鸡痘病毒转移质粒pUTAL-ORF5-ORF2。将该重组质粒与鸡痘病毒282-E4株共转染鸡胚成纤维细胞,进行同源重组。通过3次溴脱氧尿嘧啶核苷(BrdU)加压筛选,经RT-PCR、IFA和Western blot鉴定,表明重组鸡痘病毒中的目的基因在鸡胚成纤维细胞中得到表达,获得一株携带有目的基因ORF2和ORF5的重组鸡痘病毒rFPV-ORF2-ORF5。

关键词： PRRSV ORF5 pcv-2 ORF2 鸡痘病毒

华东地区PMWS患病猪群pcv-2的检测及其基因特征

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中国兽医学报 2004年第3期24卷 240-242页

作者：王先炜姜平李玉峰张书霞许家荣南京农业大学农业部动物疫病诊断与免疫重点开放实验室江苏南京210095

用 PCR方法 ,对江苏、上海、山东、江西、安徽和浙江等省市 6 5家猪场 99例断奶仔猪多系统衰弱综合征(PMWS)肺脏和淋巴结病料 ,进行了猪圆环病毒 - 2 (pcv- 2 )基因检测 ,发现其中 18家猪场 2 1例病料为 pcv- 2阳性。源自安徽和上海的 P... 详细信息

用 PCR方法 ,对江苏、上海、山东、江西、安徽和浙江等省市 6 5家猪场 99例断奶仔猪多系统衰弱综合征(PMWS)肺脏和淋巴结病料 ,进行了猪圆环病毒 - 2 (pcv- 2 )基因检测 ,发现其中 18家猪场 2 1例病料为 pcv- 2阳性。源自安徽和上海的 pcv- 2分离株经基因序列分析显示 ,2个分离株部分 ORF2 与国外分离毒株同源性为 92 %～99%。安徽分离株 AHHF6与韩国分离株 KSY- 1同源性为 99% ,而上海分离株 SHHT1与加拿大分离株 Imp.114 7同源性为 98%。上述结果表明 ,我国 PMWS患病猪群广泛存在 pcv- 2 ,不同地区 pcv- 2存在一定基因差异。

关键词：华东地区 PMWS 患病猪群 pcv-2 基因猪圆环病毒 PCR

云南地区2008～2011年部分猪场CSF、pcv-2、PRRS、PR流行情况调查与分析

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饲料工业 2013年第7期34卷 63-64页

作者：宋春莲李文贵尹革芬谭丽勤王艳芬李箐刘洁舒相华云南农业大学动物科学技术学院云南昆明650201 云南蒙自县动物疫病预防控制中心云南蒙自661100 云南普洱市思茅区动物卫生监督所云南普洱66500 云南呈贡县洛龙街道办事处云南呈贡650500

为了解云南地区部分猪场CSF、pcv-2、PRRS和PR发病情况,设计引物对291个猪场2008~2011年送检的979份组织病料进行PCR检测。结果显示,这4种病在猪场都有发生,PRRS检出率最高为55.88%,CSF、pcv-2、PR分别为30.02%、36.57%、44.91%,混合感... 详细信息

为了解云南地区部分猪场CSF、pcv-2、PRRS和PR发病情况,设计引物对291个猪场2008~2011年送检的979份组织病料进行PCR检测。结果显示,这4种病在猪场都有发生,PRRS检出率最高为55.88%,CSF、pcv-2、PR分别为30.02%、36.57%、44.91%,混合感染检出率为64.45%,单独感染占35.24%;二重感染占64.76%;三重感染占21.55%;四重感染占2.66%,二重感染以CSF＋PRRS最多,占15.24%,三重感染以CSF＋PR＋PRRS为主,占8.08%。说明在云南地区部分猪群中的这4种疫病感染比较严重,以混合感染为主,PRRS是主要的致死疫病,年度发生和流行规律变化大,应加强监督和防控工作,减少猪群呼吸道和繁殖障碍疫病的发生。

关键词：云南地区 CSF pcv-2 PRRS PR