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选取水泡性口炎病毒 N基因序列 ,设计 1对引物 ,建立检测水泡性口炎病毒的 RT- PCR方法。对VSV各毒株进行检测 ,结果均为阳性 ,而对反刍动物病毒性疾病相关病毒进行检测 ,结果均为阴性。结果表明所建立的 RT- PCR技术可用于水泡性口炎的诊断和流行病学调查 相似文献
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0.05MpH9.6碳酸盐缓冲液和0.05MPH13NaOH溶液作为包被液,分别用于间接ELISA检测乙型五号病病毒抗体。结果表明,用0.05MPH13NaOH溶泡包被可完全灭活五号病病毒抗原而用0.05MPH9.6碳酸盐缓冲液却有87%的五号病病毒抗原未被杀灭;用于检测时,以0.05MPH13NaOH溶液为包被液在敏感性及检测结果的梯度方面都优于用0.05MPH9.6碳酸盐缓冲液。 相似文献
25.
首次应用XXG-A型心血管功能测试仪以无创伤检测技术对16例妊娠黄牛22项血液动力流变学指标做了检测,并首次对怀孕黄牛的血液动力流变学特征做了报道。 相似文献
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N. J. M. Roozen J. W. L. Van Vuurde 《European journal of plant pathology / European Foundation for Plant Pathology》1991,97(5):321-334
Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation ofClavibacter michiganensis subsp.sepedonicus (Cms) from cattle manure slurry containing c. 108 colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGMI consisted of YGM supplemented with nalidixic acid (2 mg/l), polymyxin B sulphate (30 mg/l) and S-0208 (125 mg/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4×103 and 2.3×104 cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (=16 mm), proved suitable for detection, but was c. 30 times les sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R
2=0.74). 相似文献
27.
用组织培养技术将富士苹果茎尖外植体建立在MS培养基中,形成试管苗。取4—6周龄的茎与愈伤组织,徒手切取约1mm厚的横切面,印迹在硝化纤维膜上。印迹组织经封闭后,与CLSV碱性磷酸酶标抗体反应。对反应结果进行显色。感染CLSV的印迹组织呈紫色反应,正常组织无显色反应。结果表明,该方法十分容易检测印迹组织中的CLSV。带毒愈伤组织较茎的显色反应敏感,显色反应时间仅为茎的一半(15min)。CLSV在愈伤组织中有两种分布情况:一是所有组织均有显色反应;二是呈不均匀分布。CLSV在茎组织中有三种分布情况:第一,除髓部外,表皮、皮层及维管系统中均有分布;第二,不均匀分布,CLSV集中分布于表度组织中,在皮层及韧皮部仅有少量分布;第三,“嵌合”分布,即同一种组织中部分组织带毒,部分为正常组织。 相似文献
28.
柑橘裂皮病类病毒的一步RT-PCR法检测及其在甜橙体内的分布 总被引:3,自引:0,他引:3
柑橘裂皮病是由柑橘裂皮病类病毒(Citrusexocortisviroid,CEVd)引起的一种重要的柑橘病害。分别采用快速微量核酸提取法和SDS-KAc抽提法在表现症状的Etrog香橼中提取核酸,采用一步RT-PCR法对CEVd进行检测,结果显示SDS-KAc法可以消除带毒样品中非特异性条带。从嫁接接毒的铜水72-1锦橙植株的接穗部取嫩叶、老叶、皮,从砧木部茎杆上取老皮以及根皮分别提取总核酸进行RT-PCR检测,分析CEVd在甜橙体内的分布情况。初步检测结果表明在接穗的嫩叶、老叶、皮,砧木部茎杆的老皮和根皮上都可以检测到CEVd,以接穗部叶和皮检出稳定性高。 相似文献
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M. Bertamini M. S. Grando K. Muthuchelian N. Nedunchezhian 《Physiological and Molecular Plant Pathology》2002,61(6):349
We have studied the effect of the apple proliferation phytoplasmal infection on some features of the thylakoids from field grown apple (Malus pumila) leaves. Changes in photosynthetic pigments, soluble proteins, ribulose-1,5-bisphosphate carboxylase, nitrate reductase, photosynthetic activities and thylakoid membrane proteins were investigated. The level of total chlorophyll and carotenoids were reduced in phytoplasma-infected leaves. Similar results were also observed for soluble proteins and ribulose- 1,5-bisphosphate carboxylase activity. The in vivo nitrate reductase activity was significantly reduced in infected leaves. When various photosynthetic activities were followed in isolated thylakoids, phytoplasmal infection caused marked inhibition of whole chain and photosystem II activity while the inhibition of photosystem I activity was only marginal. The artificial exogenous electron donors, diphenyl carbazide and hydroxylamine significantly restored the loss of photosystem II activity in infected leaves. The same results were obtained when Fv/Fm was evaluated by chlorophyll fluorescence measurements. The marked loss of photosystem II activity in infected leaves could be due to the loss of 47, 33, 28–25, 23 and 17 kDa polypeptides. It is concluded that phytoplasmal infection inactivates the donor side of photosystem II. This conclusion was confirmed by immunological studies showing that the content of the 33 kDa protein of the water-splitting complex was diminished significantly in infected leaves. 相似文献