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为建立可同时检测猪捷申病毒(PTV)与猪圆环病毒2型(PCV2)的双重PCR方法,本研究根据GenBank登录的相关病毒基因序列,选择保守区域设计引物,经过反应条件的优化,建立了可同时检测以上2种病毒的PCR诊断方法,扩增片段长度分别为187bp(PTV)、120bp(PCV2)。通过实验证明该方法具有良好的特异性和较高的敏感性,对PTV、PCV2核酸检测最低浓度分别为2.8×10^-2ng和6.6×10^-3ng。应用该方法对43份临床样品进行检测发现,PTV阳性率为23%,PCV2阳性率为38%,PTV与PCV2共感染率为16%。该方法的成功建立,为快速高效地检测以上2种病毒提供有力手段。 相似文献
44.
Rebecca J. Kessler Shelley Rankin Sheri Young Kathleen O'Shea Maria Calabrese Amy Guldin Nicole Lipson Donna A. Oakley Urs Giger 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2010,39(1):29-38
Background: While screening programs have reduced the risk of infectious disease transmission by donors in human and veterinary blood banking, bacterial contamination of blood products has emerged as a major complication in human medicine. Objectives: To describe a Pseudomonas fluorescens (Pf)‐contaminated feline packed RBC (pRBC) unit and experimentally investigate Pf‐contaminated canine pRBCs. Methods: Canine pRBCs were inoculated with Pf‐rich pRBCs from the sentinel feline unit and stored at 4°C or 20°C for 72 hours. Aliquots from the pRBCs were serially evaluated by microscopy, culture, and a eubacterial 16S rRNA real‐time PCR assay. Results: One Pf‐contaminated feline unit turned black after 22 days of storage and was removed from the blood bank; a source was not found, and no other contaminated units were identified. Canine pRBCs spiked with 5 or 25 μL of the sentinel unit became culture‐ and/or 16S PCR‐positive at ≥8 hours at 20°C and 48 hours at 4°C and developed a color change at ≥24 hours. Sensitivity studies indicated that without incubation, inoculation of ≥100 μL Pf‐rich pRBCs was necessary for a positive 16S PCR test result. Conclusions: P. fluorescens grows in stored pRBCs slowly at 4°C and rapidly at 20°C. Screening of blood products for color change, estimating bacterial concentration with microscopy, and 16S PCR testing are simple and fast ways to detect bacteria in stored blood. Aseptic collection, temperature‐controlled storage, and regular visual monitoring of stored units is recommended. Discolored units should not be transfused, but examined for bacterial contamination or other blood product quality problems. 相似文献
45.
新疆地区规模化奶牛场牛支原体流行病学调查 总被引:2,自引:0,他引:2
为了调查新疆地区规模化奶牛场牛支原体的感染情况,采用牛支原体间接ELISA和特异性PCR检测牛血清中的牛支原体抗体及病料中牛支原体核酸,共检测9个地区15个规模化奶牛场437份血清,肺脏、关节液及鼻腔黏液44份。结果显示,血清抗体阳性率为76.43%(334/437),其中脐带血抗体阳性率为40.00%(4/10);病料阳性率为40.91%(18/44)。检测结果表明,新疆地区大部分奶牛场存在牛支原体感染,部分奶牛场发生牛支原体肺炎及关节炎病例,并存在垂直传播的风险。牛支原体感染可能成为危害新疆规模化奶牛场犊牛健康的主要疫病之一。 相似文献
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Expression of receptor tyrosine kinases VEGFR-1 (FLT-1), VEGFR-2 (KDR), EGFR-1, PDGFRa and c-Met in canine primary brain tumours 总被引:1,自引:0,他引:1
P. J. Dickinson B. N. Roberts R. J. Higgins C. M. Leutenegger A. W. Bollen P. H. Kass R. A. LeCouteur 《Veterinary and comparative oncology》2006,4(3):132-140
Inhibition of tumour growth and angiogenesis by targeting key growth factor receptors is a promising therapeutic strategy for central nervous system tumours. Characterization of these growth factor receptors in canine primary brain tumours has not been done. Using quantitative real‐time TaqMan polymerase chain reaction (PCR), we evaluated the expression of messenger RNA (mRNA) for five tyrosine kinase growth factor receptors (vascular endothelial growth factor receptor [VEGFR]‐1, VEGFR‐2, endothelial growth factor receptor [EGFR]‐1, platelet‐derived growth factor receptor a [PDGFRa], and c‐Met) relative to normal cerebral cortex in 66 spontaneous canine primary brain tumours. Increased expression of VEGFR‐1 and VEGFR‐2 mRNA was greatest in grade IV astrocytomas (glioblastoma multiforme) and grade III (anaplastic) oligodendrogliomas. EGFR‐1 mRNA expression was more consistently increased than the other receptors in all tumour types, while increased PDGFRa mRNA expression was mostly restricted to oligodendrogliomas. The similarities in increased expression of these tyrosine kinase growth factor receptors in these canine tumours, as compared to data from their human counterparts, suggest that common molecular mechanisms may be present. 相似文献
48.
YANG Yang XU Hou-qiang CHEN Wei ZHOU Di XU Min ZHANG Qing-qing ZHAO Huan-ping SUN Cheng-juan WANG Yuan-yuan ZHANG Ming YANG Tao 《中国畜牧兽医》2017,44(12):3401-3409
The objective of this study was to clone PDK4 and FGF10 genes, and investigate the expression level of PDK4 and FGF10 genes mRNA in different tissues of Large White pig and Congjiang Xiang pig. The PDK4 and FGF10 genes were cloned by RT-PCR and analyzed by bioinformatics, the relative expression of PDK4 and FGF10 genes were detected by Real-time PCR. The results showed that the coding region of PDK4 gene was 1 224 bp, encoding 407 amino acids; The coding region of FGF10 gene was 636 bp and encoded 211 amino acids. The homologies of nucleotide sequences of PDK4 gene with sheep, horse and human were 93%, 92% and 91%,respectively. The homologies of nucleotide sequences of FGF10 gene with sheep, cattle, human and mouse were 94%,93%, 93% and 90%, respectively. The phylogenetic tree of PDK4 gene showed that the genetic relationship of Congjiang Xiang pig, cattle and sheep were very close, the phylogenetic tree of FGF10 gene indicated that the genetic relationship of Congjiang Xiang pig, cattle, sheep, human and macaque were very close, but the genetic relationship of Congjiang Xiang pig, rat and chicken were far away. Real-time PCR results showed that, in different tissues of Congjiang Xiang pig,PDK4 gene expression in kidney tissue was higher than other tissues, with a higher expression in stomach and adipose as well,FGF10 gene expression in stomach tissue was higher than other tissues, with a higher expression in kidney and adipose as well, but both of PDK4 and FGF10 genes expression were the lowest in longissimus dorsi. In different tissues of Large White pig, both of PDK4 and FGF10 genes were expressed the highest in adipose than other tissues, PDK4 gene expression in longissimus dorsi was the lowest, while the FGF10 gene expression the lowest in heart. This study successfully cloned the PDK4 and FGF10 genes of Large White pig and Congjiang Xiang pig,and detected the relative expression of PDK4 and FGF10 genes in different tissues of Large White pig and Congjiang Xiang pig, and also provided scientific basis for further study on regulation of PDK4 and FGF10 genes on lipid metabolism and deposition. 相似文献
49.
杨盘二孢激发子的分离及稳定性研究 总被引:1,自引:0,他引:1
从杨盘二孢的培养滤液和菌丝中获得2种激发子粗提物,分别测定其糖和蛋白质的含量,发现滤液激发子粗提物(CFE)糖和蛋白质的含量分别为41.07和40mg/mL,菌丝激发子粗提物(CME)糖和蛋白质含量分别为48.07和55mg/mL。滤液激发子粗提物对温度不敏感而对碱性条件敏感;菌丝激发子粗提物对酸碱不敏感而对温度敏感。用Sephadex G-100柱层析的方法初步纯化2种激发子,并且菌丝激发子粗提物过柱后得到2个活性组分J14和J25;滤液激发子粗提物过柱后获得2个活性组分L9和L16。将4个组分进行烟草叶片过敏反应和I-895杨酶活性的诱导,结果表明,菌丝激发子强活性物质集中在J14组分;而滤液激发子强活性物质集中在L9组分。 相似文献
50.
采用培养皿法和常规生化方法,研究了新型除草剂唑嘧氯草胺(暂定名,代号:ZJ-2725)的作用机制。结果显示,同时添加20mg/L浓度的缬氨酸、亮氨酸和异亮氨酸能完全恢复唑嘧氯草胺对苘麻芽的生长抑制作用,而添加相同浓度的单一支链氨基酸只能部分消除其抑制作用。离体条件下,随着唑嘧氯草胺浓度的增大,对乙酰乳酸合成酶(ALS)活性的抑制率增加,ALS活性随反应时间的延长而增加,且两者之间符合Michaelis-Meten方程;活体条件下,唑嘧氯草胺对苘麻的ALS也表现出明显的抑制作用,ALS活性明显下降。表明唑嘧氯草胺在植物体内抑制了3种支链氨基酸的生物合成,进而导致植物蛋白质合成受阻而使植物生长受到抑制,ALS即为唑嘧氯草胺的作用靶标。 相似文献