排序方式： 共有80条查询结果，搜索用时 15 毫秒
21.
为对犬孢子虫MAG1与IFN-γ融合基因重组真核质粒进行免疫学评价,本实验以含有MAG1-IFN-γ基因片段的克隆质粒为模板,扩增MAG1-IFN-γ目的片段,构建pVAX-MAG1-IFN-γ重组真核表达质粒,将其转染Vero细胞,采用间接免疫荧光(IFA)和western blot技术检测MAG1基因在Vero细胞中的表达,并免疫BALB/c小鼠,进行免疫学评价。结果表明,PCR扩增获得基因片段大小为1 536 bp,与GenBank中登录的MAG1和IFN-γ核苷酸序列的同源性为99%;IFA检测MAG1基因在Vero细胞中获得瞬时表达,转染的Vero细胞呈现特异性绿色荧光;western blot分析表达蛋白的分子量为57 ku,具有较好的反应原性。将构建的重组真核表达质粒免疫BALB/c小鼠,经间接ELISA和T淋巴细胞亚群含量测定表明,重组质粒具有较好的免疫原性。本实验为新孢子虫病核酸疫苗的深入研究奠定了基础。 相似文献
22.
Forty per cent of the members of the Norwegian Small Animal Veterinary Association (NSAVA) returned a questionnaire regarding the treatment and diagnosis of canine nasal mite infection in their practices in 1996. A total of 2392 dogs were treated for this infection by 156 NSAVA members in clinical practice in that year, averaging 15.3 treated dogs per veterinarian. An estimated minimum of 6000 Norwegian dogs were treated for nasal mite infection in 1996, at an estimated cost of treatment exceeding 3.3 million NOK. Fifty-nine per cent of the practitioners included at least a case history and clinical examination as diagnostic criteria prior to initiating treatment for nasal mite infection. Fewer than 10% routinely performed more thorough diagnostic procedures, and only 1.8% of the diagnoses were verified prior to treatment. A total of 27 different treatment regimens, involving either subcutaneously injected ivermectin or orally administered milbemycin oxime, were routinely used by NSAVA veterinarians to treat these dogs. Seventy-two (53%) of the veterinarians used ivermectin exclusively, 14 (10%) used milbemycin oxime exclusively, while 50 (37%) used both ivermectin and milbemycin oxime. No other drugs were used. Evaluation of treatment was made by assessing resolution of the clinical signs following treatment, and 97% of the veterinarians were satisfied with the effect of the treatment regimen they used. Adverse side-effects following treatment for nasal mite infection in 1996 were seen on 9 occasions; 4 following ivermectin administration and 2 following milbemycin oxime treatment, while in 3 cases no information on the drug used was provided. The risk of adverse reactions to treatment of nasal mite infection in dogs was 0.4%. 相似文献
23.
根据犬新孢子虫NcSAG1-NcSRS2融合基因序列,设计了1对含有Kozak序列、终止密码子、BamHⅠ和XhoⅠ酶切位点的引物,以含有NcSAG1-NcSRS2融合基因的质粒pGEX-tNcP43-P36为模板,经PCR扩增获得NcSAG1-NcSRS2融合基因片段,用BamHⅠ和XhoⅠ双酶切该片段,回收得到含有以上2个酶切位点黏端的NcSAG1-NcSRS2融合基因,将此基因片段克隆至相同酶切回收后的pcDNA3.1（＋）真核表达载体中,获得重组质粒pcNCSAG1-SRS2。经PCR鉴定、限制性内切酶酶切分析和克隆片段序列测定、比较,证实了重组质粒的正确性。将构建好的真核表达质粒转染到COS-7细胞中进行瞬时表达,经免疫荧光检测,证实了该载体能在细胞内进行蛋白表达。 相似文献
24.
新孢子虫NcSRS2基因的亚克隆和表达 总被引：1，自引：0，他引：1
本研究根据NcSRS2基因序列设计合成一对引物,将上、下游引物分别引入EcoRI,XhoI酶切位点,用PCR技术从pGEX-NcSRS2重组质粒扩增截去N端疏水氨基酸序列NcSRS2的基因片段（以下称dNcSRS2）,插入到pGEX-6p-1质粒的多克隆位点,转化大肠杆菌BL21感受态细胞,于氨苄阳性LB培养平板上筛选阳性克隆,酶切及PCR鉴定;经IPTG诱导在E.coli中表达,用SDS-PAGE和免疫印迹分析表达产物并纯化.结果表明,新孢子虫dNcSRS2基因体外扩增产物与预期值相符,约1041bp;所构建pGEX-dNcSRS2重组质粒经双酶切与PCR鉴定,与预期结果一致;SDS-PAGE和免疫印迹显示,表达融合蛋白的分子量约为62.6 kD,表达效率为32.3%,该蛋白具有特异的免疫反应性,为新孢子虫病诊断试剂盒的研制和疫苗的研制奠定了基础. 相似文献
25.
将构建成功的重组质粒pVAX-SAG1转染至BHK21细胞上,通过IFAT与RT-PCR方法检测重组质粒在BHK21细胞上的表达情况。大量提取重组质粒pVAX-SAG1,免疫Balb/c小鼠,分别在首免前、二免前、三免前以及三免后的第1、2、3周进行小鼠断尾采血,应用ELISA方法与流式细胞技术检测体液免疫水平和细胞免疫水平。结果表明,转染的重组质粒pVAX-SAG1在BHK21细胞中得到瞬时表达,并且重组质粒pVAX-SAG1免疫Balb/c小鼠后,在体液免疫水平上,pVAX-SAG1组与空载体pVAX1和PBS对照组差异极显著（P〈0.01）;在细胞免疫水平上,pVAX-SAG1组与PBS对照组差异显著（P〈0.05）。本试验为新孢子虫基因工程核酸疫苗的进一步研究奠定了基础。 相似文献
26.
为了观察米尔贝肟片对犬弓蛔虫和钩虫驱虫效果,采用人工感染方式,对犬弓蛔虫和钩虫的动物模型进行疗效研究。结果表明,单次口服0.5 mg/kg米尔贝肟片对犬弓蛔虫和犬钩虫的虫卵减少率、驱虫有效率和治愈率均可达到100%。说明米尔贝肟片在治疗犬弓蛔虫和钩虫时具有剂量小、疗效高、作用快、副反应轻等特点,在临床治疗方面有十分广阔的应用前景。 相似文献
27.
28.
Systemic neosporosis in a dog treated for immune‐mediated thrombocytopenia and hemolytic anemia 
下载免费PDF全文
下载免费PDF全文 Angie Magaña Félix Sánchez Karina Villa Liliana Rivera Elizabeth Morales 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2015,44(4):592-596
A 4‐year‐old male Toy Poodle was presented to the Small Animal Veterinary Hospital of the Faculty of Veterinary Medicine of the Autonomous University of Mexico (FMVZ, UNAM) because of depression, lethargy, and hemorrhages involving several areas of the skin and around the eyes. Hematology data and a bone marrow analysis suggested hemolytic anemia and immune‐mediated thrombocytopenia. The dog was treated with prednisone, and after one month the hematology variables improved. However, the dog's clinical condition inexplicably worsened and it was euthanized. On necropsy, there were no relevant findings. However, in histology, multifocal lymphoplasmacytic and histiocytic meningoencephalitis and necrosis, and a protozoan cyst in the cerebellum were identified. In addition, moderate multifocal lymphoplasmacytic and necrotizing pancreatitis, hepatitis, myocarditis, and diffuse lymphoplasmacytic enteritis were observed. Immunohistochemistry of the cerebellum, liver, pancreas, and intestine with a specific antibody against Neospora caninum confirmed the diagnosis of systemic neosporosis. The systemic neosporosis in this dog was most likely caused by reactivation of latent parasites due to prednisone administration during the one month of treatment. It should be kept in mind that in dogs being treated with immunosuppressants for immune‐mediated conditions, opportunistic parasites, such as Toxoplasma gondii and N caninum, can be reactivated from a latent state, as it probably happened in the present case. 相似文献
29.
为了解新孢子虫AMA1基因的生物信息学特性,并构建重组原核表达质粒p GEX-4T-1-Nc AMA1。本试验对牛源犬新孢子虫吉林株AMA1基因进行分子克隆,利用生物信息学软件对该基因进行编码蛋白等电点、信号肽、跨膜区、糖基化位点及疏水性分析,并将该基因片段亚克隆至原核表达载体p GEX-4T-1,构建了重组表达质粒p GEX-4T-1-Nc AMA1。结果显示,克隆的Nc AMA1基因片段长1 695 bp,与GenBank(AB265823.1)上发表的基因序列同源性99.9%;Nc AMA1基因编码蛋白等电点为5.43,在其N端及C端分别存在一个跨膜区,为不溶性蛋白,且Nc AMA1蛋白抗原指数较高,有潜在疫苗研究价值。 相似文献
30.
AIM: To estimate the distribution and causes of abortion in dairy herds across New Zealand during the 2001/2002 breeding season. METHODS: A questionnaire survey was sent to all veterinary practices that employed members of the New Zealand Veterinary Association. Veterinarians were requested to record the numbers of dairy herds serviced by their practice in which the rate of aborting cows was zero (0%), low (1–5%) and high (6–10% and >10%), and all laboratory submissions, specimens, tests and results related to their investigation of abortions in these herds, for each category. RESULTS: Forty-two large animal practices serving 1,431 dairy clients from all except one region in New Zealand responded. The within-herd incidence of abortion was zero in 497 herds (34.7%), low in 876 herds (61.2%), and high in 58 herds (4.1%) in the season 2001/2002. Within the high category, 0.6% herds reported abortions at a frequency as high as may be expected in epidemic abortions (>10%). Compared to other regions, the abortion incidences tended to be relatively high in Northland, Auckland and the Bay of Plenty, and low in Hawke Bay, Waikato and Nelson (p<0.001). Cases were submitted for laboratory investigation from 51/934 (5.5%) herds. Even though submission rates increased with the level of abortion, most submissions originated from herds with a low abortion rate. The main agents identified among submissions as the cause of abortion were Neospora caninum (Nc) (35%) and bovine viral diarrhoea virus (BVDV) (16%). Among 39 submissions from low abortion herds, 33% were suspected due to Nc and 15% to BVDV, and this was not statistically different (p>0.10) from the distribution in 13 submissions from high abortion herds (46% Nc, 23% BVDV). While cases associated with Nc were submitted in April, May and June, submissions for suspected BVDV spread evenly over the months of March to July. CONCLUSION: In the 2001/2002 season, approximately 4% of dairy herds in New Zealand recorded a relatively high incidence of abortion (>5%) and 0.6% herds experienced an incidence of >10%. BVDV and Nc were the most frequently diagnosed agents associated with abortion, regardless of whether the seasonal incidence of abortion within a herd was low or high. 相似文献