猪圆环病毒2型/3型双重荧光定量PCR检测方法     

王林青  樊霖  赵宇  田润博  崔建涛  韩昊莹  赵丽  陈红英 《安徽农业大学学报》2021,48(4):595-599

为了能够同时快速地检测和鉴别猪圆环病毒2型和3型(PCV2和PCV3),参考GenBank中已发表的PCV2和PCV3基因组序列,针对其保守区分别设计了2对特异性引物,经优化反应体系和条件,建立了能快速检测和鉴别PCV2/PCV3双重荧光定量PCR方法.结果 表明,PCV2和PCV3的R2分别为0.999、0.9993,E值分别为3.5731、3.3734.该方法能同时特异地检测PCV2和PCV3,而对其他5种猪病原均未检测到荧光信号;PCV2和PCV3的最低检测值分别为41.1 copies·μL-1、27.0 copies·μL-1;批内和批间变异系数均小于1％.临床样本检测结果显示,PCV2和PCV3的阳性率分别为62.12％(41/66)、48.48％(32/66),二者混合感染的阳性率为46.96％(31/66).表明该方法具有敏感、特异和可靠等特点,该方法为PCV2和PCV3单独或者混合感染的早期诊断、定量检测及其流行病学调查提供了可行的技术支持.  相似文献   

代乳料原料组成对早期断奶羔羊生长及脏器发育的影响     

李典芬  张鹏坤  魏时来  李伟  黄殿选  陈颢  李桂英  赵玉军  王志鹏 《草业科学》2010,27(8):129-133

为了探讨代乳料原料的不同组合效应对早期断奶羔羊的饲喂效果,选取25只甘肃高山细毛羔羊,分为5组,每组5只,对照组羔羊由母羊自由哺乳和放牧;试验组羔羊30日龄断奶,饲喂4种代乳料:A组主原料未膨化,B组主原料膨化,C组主原料膨化+血浆蛋白粉,D组在C组基础上增加乳清粉。试验期为30~90日龄。结果表明:50、70日龄时,对照组羔羊体质量最大,试验组羔羊受断奶应激的影响,相应指标较小。90日龄时,D组体质量显著高于B、C组(P0.05),超过了对照组。30~90日龄阶段,D组日增体质量最大,且显著高于B、C组(P0.05)。对照组脾脏质量显著高于4个试验组(P0.05),且脾脏指数最高。说明在代乳料中添加血浆蛋白粉及增加乳清粉,有利于羔羊健康发育,使代乳料的饲喂效果最佳。  相似文献   

Attenuation of Virulent Porcine Reproductive and Respiratory Syndrome Virus Strain CH-la and Genetic Variation of ORF5 Gene     

CAI Xue-hui  WU Guo-jun  LIU Yong-gang  LIU Guang-qing  SHI Wen-da  WANG Shu-jie  MAPing  LI Cheng-jun  HAN Wen-yu 《中国农业科学(英文版)》2012,(12):2035-2042

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Demonstration of Heterogeneous Genotypes of Taylorella equigenitalis Isolated from Horses in Six European Countries by Pulsed-Field Gel Electrophoresis     

Kagawa S  Klein F  Corboz L  Moore JE  Murayama O  Matsuda M 《Veterinary research communications》2001,25(7):565-575

Forty-six isolates of Taylorella equigenitalis were analysed by pulsed-field gel electrophoresis (PFGE) after separate digestion of the genomic DNA with ApaI and with NotI. The isolates had been obtained from horses in six European countries and were classified into 18 genotypes. In Belgium, 2 genotypes were detected in 2 isolates, in England 9 among 15, in Finland 2 in 2, in France 2 among 10, in Sweden 3 among 5, and in Switzerland 3 among 12. Two English isolates and 4 French isolates gave identical PFGE profiles to those of Kentucky 188 from the United States. A common genotype was found in 5 isolates from Belgium and England and also in 10 isolates from France and Switzerland. The analysis of genomic DNA from 12 isolates of T. equigenitalis obtained from male horses in France, Sweden and Switzerland gave no evidence of a sex-related difference in the genomic DNA. Genomic DNA from 11 streptomycin (STM)-susceptible isolates obtained in Sweden and Switzerland were classified into four genotypes by PFGE. Each of the six genotypes determined among the 17 isolates from these two countries had single phenotypes for resistance or susceptibility to STM.  相似文献   

Double in situ hybridization for simultaneous detection and differentiation of porcine circovirus 1 and 2 in pigs with postweaning multisystemic wasting syndrome   总被引：4，自引：0，他引：4  

Kim J  Chae C 《Veterinary journal (London, England : 1997)》2002,164(3):133-253

Double in situ hybridization using a digoxigenin-labelled porcine circovirus 1 (PCV1) and biotinylated PCV2 probe, was developed for the simultaneous detection and differentiation of PCV1 and PCV2 in formalin-fixed, paraffin-embedded tissues from pigs with postweaning multisystemic wasting syndrome. The combination of an alkaline phosphatase conjugated antidigoxigenin system with alkaline phosphatase conjugated streptavidin-biotin system allowed identification of PCV1 and/or PCV2. No evidence of cross-reaction was observed. Positive cells exhibited a red or dark brown reaction product for PCV1 and PCV2, respectively. Both PCV DNAs were observed mainly in the cytoplasm but occasionally in the nucleus. Co-localization of hybridization signal for both PCV1 and PCV2 was present in macrophages and multinucleated giant cells of the lymph node and spleen. This double-labelling technique for the differentiation between PCV1 and PCV2 is suitable for pathogenesis studies and diagnostic applications.  相似文献   

猪脑心肌炎病毒VP1白单克隆抗体的制备及鉴定     

《中国兽医科技》2007,37(1):24-28

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Genetic aspects concerning drip loss and water-holding capacity of porcine meat     

D. G. J. Jennen  A. D. Brings  G. Liu  H. Jüngst  E. Tholen  E. Jonas  D. Tesfaye  K. Schellander  & C. Phatsara 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2007,124(S1):2-11

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Isolation and Identification of a Variant Porcine Epidemic Diarrhea Virus and the Immune Efficacy of Its Inactivated Vaccine     

CHANG Xianyun  ZHAO Runze  TONG Ke  CHEN Shaoxian  LIU Guoping 《中国畜牧兽医》2007,47(11):3651-3658

This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID₅₀,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×10^8.0 TCID₅₀/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×10^8.0TCID₅₀/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China.  相似文献   

小鼠4、8-细胞胚胎单卵裂球体外培养体系的研究     

胡辛怡  周光斌  杨中强  刘国世  朱士恩 《中国畜牧杂志》2006,42(11):12-15

以CZB为基础培养液,培养小鼠4、8-细胞胚胎单卵裂球,研究葡萄糖、牛磺酸和猪输卵管上皮细胞共培养在其体外发育中的作用。结果表明:牛磺酸添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为29%、30%和14%、14%,无显著性差异(P>0.05)。添加葡萄糖后,4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%和19%,有显著提高(P<0.05)。含有葡萄糖而牛磺酸的添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%、38%和19%、20%,无显著性差异(P>0.05)。各组内4、8-细胞胚胎单卵裂球形成的囊胚细胞数分别为(10.44±1.24～(12.43±1.18)和(7.57±0.97)～(8.48±1.16),均无显著性差异(P>0.05)。4、8-细胞胚胎单卵裂球与猪输卵管上皮细胞共培养,其囊胚发育率分别为47%和26%,囊胚细胞数分别为(17.57±1.13)和(11.43±0.92),均高于单一培养(P<0.05)。  相似文献   

PCV2感染性分子克隆的制备及体内外感染性试验     

韩凌霞  刘建华  陈艳  仇华吉  王云峰  童光志 《中国预防兽医学报》2006,28(3):241-247,270

将猪圆环病毒2型（Type2 porcine circovirus，PCV2）JXL株cDNA插入到载体pMD18-T中，获得的pMDT-PCV转染猪肾细胞PK15细胞系，盲传4代后，利用PCV2阳性猪血清进行间接免疫荧光抗体试验（IFA），能检测到特异性荧光。将pMDT-PCV质粒DNA腹股沟淋巴结注射40日龄仔猪2头，分别接种2、3次，接种剂量500μg／次，首次注射35d心脏放血致死。在体内感染性试验期间，动物未出现明显临床症状。试验结束，病理切片结果显示，不同淋巴组织中分别存在淋巴细胞缺失或增多的现象；肾小球、扁桃体、空肠淋巴结和股前淋巴结等冰冻组织切片中存在大量PCV2病毒抗原；血清中PCV2特异性的ELISA抗体效价迭1：2560，IFA效价为1：1280；通过聚合酶链式反应（PCR）可以分别从体外感染PK15细胞和体内感染动物及同圈饲养的空白对照猪的淋巴组织中扩增到病毒特异性基因片段。本研究证明含有单拷贝PCV2的重组质粒pMDT—PCV在体内、外均具有感染性，能衍生出病毒，并产生符合自然感染PCV2的大体病变和组织病理学变化，并激发较强的体液免癌应答。  相似文献