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牙Ping淋巴囊肿病的PCR诊断方法研究 总被引:2,自引:0,他引:2
以中国养殖牙Ping(Paraclichthys olivaceus)淋巴囊肿病毒(Lymphocystis disease virus,LCDV_cn)主要衣壳蛋白(major capsid protein,MCP)基因的中间保守序列为目标基因,设计了一对特异性引物。该引物可扩增出172bp的病毒DNA片段,其最小DNA检出量为0.0183ng。用PCR法从人工感染淋巴囊肿病毒3天的牙Ping血,鳃,肝,脾,肠,胃及自然发病牙Ping的肿瘤中,分别检测到了LCDV的存在。本实验结果证明,PCR法对于早期检测LCDV是十分有效的。 相似文献
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军曹鱼淋巴囊肿病毒的系统关系研究及基因型分析 总被引:1,自引:0,他引:1
从患淋巴囊肿病的养殖军曹鱼肿瘤中分离到一种虹彩病毒,命名为淋巴囊肿病毒军曹鱼分离株(LCDV-rc).为探讨LCDV-rc的系统地位,从DNA水平上分析了该病毒株与淋巴囊肿病毒中国牙鲆分离株(LCDV-cn)、淋巴囊肿病毒韩国鲈鱼分离株(LCDV-sb)、淋巴囊肿病毒日本牙鲆分离株(LCDV-jf)、淋巴囊肿病毒欧洲分离株(LCDV-1)和淋巴囊肿病毒日本岩鱼分离株(LCDV-rf)的系统关系.以核衣壳蛋白(MCP)基因为分子标记,通过PCR扩增和克隆测序获得了长度为1356bp的含178个信息位点的mcp基因序列;用最大简约法、最大似然法、邻接法和Bayesian法构建了分子系统树并进行了置信度检验.结果表明,构建的淋巴囊肿病毒ML、NJ、MP和Bayesian系统树的拓扑结构一致,LCDV-rc和LCDV-sb聚为一支,LCDV-cn和LCDV-jf聚为一支,LCDV-1和LCDV-rf分别各为一支.在35种虹彩病毒的系统关系分析中,LCDV-rc与另5株淋巴囊肿病毒在系统树上聚为一大支,说明LCDV-rc与淋巴囊肿病毒的关系近于其他虹彩病毒.根据遗传距离和系统关系分析认为,LCDV-rc属于一种新的基因型,称其为淋巴囊肿病毒Ⅳ型基因型. 相似文献
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牙鲆淋巴囊肿病毒在牙鲆鳃细胞系FG-9307中的增殖 总被引:9,自引:1,他引:8
向已建立的牙鲆(Paralichthys olivaceus)鳃细胞系^[1](FG-9307)中接种淋巴囊肿病(Lymphocystis disease virus,LCDV)、探讨其是否可支持LCDV的增殖。光学显微镜观察发现:接种3天后,呈纤维状的鳃细胞隆起、立体感加强;之后细胞逐渐变圆,一周左右绝大部分鳃细胞变圆、呈现了细胞病变(cytopathic effect,CPE)现象。将CPE细胞制作超薄切片,电镜观察发现细胞内有大量具囊膜、直径133-186nm的LCDV。CPE细胞的核变形、线粒体增加且空洞化,出现了与淋巴囊肿细胞相同的特征。本实验证明:FG-9307是可持续LCDV增殖的鱼类细胞系。 相似文献
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淋巴囊肿病毒核酸疫苗在牙鲆体内的表达及免疫效果评价 总被引:2,自引:0,他引:2
将本试验室得到的牙鲆淋巴囊肿病毒(LCDV-cn)衣壳蛋白0.6kb片段重组质粒pEGFP-N2-LCDV-cn MCP0.6肌注入牙鲆体内,荧光显微观察及特异性RT-PCR检测其在牙鲆组织内的表达,并进一步测定牙鲆外周血、肠、脾脏、前肾的淋巴细胞增殖反应、呼吸爆发活性及抗体产生水平.研究结果表明, pEGFP-N2-LCDV-cn MCP 0.6可分布于牙鲆各组织内,并在一定时间内持续表达;免疫学检测表明,该重组质粒诱导了牙鲆特异性体液免疫和细胞免疫.本试验成果为淋巴囊肿病毒核酸疫苗的生产应用和产业化提供了重要的理论依据. 相似文献
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牙鲆淋巴囊肿病毒主要衣壳蛋白基因片段真核表达载体的构建、表达与免疫效果评价 总被引:1,自引:0,他引:1
用淋巴囊肿病毒LCDV-cn感染牙鲆鳃细胞系FG-9307,提取细胞总RNA,用RTPCR法获得了LCDV-cn主要衣壳蛋白(MCP)0.6kb基因片段.将该LCDV-cn-MCP0.6kb片断克隆入真核表达载体pEGFP-N2,得到重组质粒pEGFP-N2-LCDV-cn-MCP 0.6.采用脂质体法将重组质粒转染入牙鲆鳃细胞系FEC,并进行瞬时表达.通过荧光显微镜观察和特异性RT-PCR检测,证实重组质粒pEGFP-N2-LCDV-cn-MCP 0.6kb已成功转染到FEC细胞,并得到了初步表达.将重组质粒肌注入牙鲆体内,检测牙鲆外周血、肠、脾脏、前肾和淋巴细胞的增殖反应、呼吸爆发活性及抗体产生水平.结果表明,构建的核酸疫苗pEGFP-N2-LCDV-cn-MCP 0.6可诱导牙鲆特异性体液免疫和细胞免疫,具有明显的免疫保护作用. 相似文献
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牙鲆淋巴囊肿病的诊断技术研究 总被引:12,自引:0,他引:12
使用本实验室设计的PCR引物,在标记反应体系下,通过PCR反应合成了碱基数为172bp的牙鲆(Paralichthys olivaceus)囊肿病(Lymphocysits disease,LCD)的地高辛标记探针,在其琼脂糖凝胶电泳图172bp处有明显的亮带,由软件分析得出,合成探针的浓度为500ng/μl,此结果证明,该产物确是所需的探针,且具有很好的合成效率。通过实验证明本文采用的核酸探针-杂交诊断方法可以有效地定性和定位LCDV。实验结果还提示。LCDV是通过血液在牙鲆体内传播的,并可能同时存在垂直和水平两种传播方式。 相似文献
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A method is proposed for estimating the parameters of a mixture of exponential and Weibull distributions for which the accuracy of preliminary estimates obtained by graphical analysis is refined in accordance with the criterion of maximum likelihood. The efficiency of the proposed method is supported by the results of statistical modeling. 相似文献
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Liebert A Wabnitz H Grosenick D Möller M Macdonald R Rinneberg H 《Applied optics》2003,42(28):5785-5792
A novel method for the determination of the optical properties of tissue from time-domain measurements is presented. The data analysis is based on the evaluation of the first moment and the second centralized moment, i.e., the mean time of flight and the variance of the measured distribution of times of flight (DTOF) of photons injected by short (picosecond) laser pulses. Analytical expressions are derived for calculation of absorption and of reduced scattering coefficients from these moments by application of diffusion theory for infinite and semi-infinite homogeneous media. The proposed method was tested on experimental data obtained with phantoms, and results for absorption and reduced scattering coefficients obtained by the proposed method are compared with those obtained by fitting of the same data with analytical solutions of the diffusion equation. Furthermore, the accuracy of the moment analysis was investigated for a range of integration limits of the DTOF. The moment analysis may serve as a comparatively fast method for evaluating optical properties with sufficient accuracy and can be used, e.g., for on-line monitoring of optical properties of biological tissue. 相似文献
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An approximate analytical expression is derived for the velocity of the front of the wave of change of boiling modes on a
rod and on a plate. The impact of the Thomson effect and of the heater orientation in the gravity field is included. Adequate
agreement is demonstrated between the derived relations and experimental data. 相似文献
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Conventional methods for summation of direct measurement errors are considered. Conditions under which these methods provide consistent results are stated. It is proposed to utilize for characteristic of the total error, its interval estimator or the half-sum of the value obtained by adding up its arithmetic and geometric random errors.Translated from Izmeritel'naya Tekhnika, No. 4, pp. 10–12, April, 1995. 相似文献
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由于空压机进气侧轴承箱蜗壳温度探头密封效果不好,使空压机及其后续管道受到油污染。后将温度探头的O形圈密封改为骨架油封,杜绝了轴承箱漏油问题。文章简介空压机的结构和技术参数,详细介绍了骨架油封的制作和优点。 相似文献
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M. M. Gadenin 《Inorganic Materials》2013,49(15):1352-1356
It is shown that conditions of attainment of marginal states in the material of structural components are formulated on the basis of analysis of force and deformation criteria of formation and development of destruction. The parameters of these criteria are data on types of structural materials, standard and special characteristics of their mechanical properties, structural forms of considered elements of the equipment, and their operating stress loading. In this case, the construction of a system of state equations for describing interrelation of current damage, external actions, and responses to them is based on the concept of calculating variations in properties of materials at all stages of the design life cycle. 相似文献
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V. T. Kornienko 《Strength of Materials》1991,23(7):760-765
The article suggests a method of evaluating the microstructural inhomogeneity of deformation of metals by modeling. The method includes the measurement of deformations of metal according to meshes of a dividing grid with small base and the calculation of the distribution parameters of microdeformations in model representation. It is shown that the modeling method makes it possible objectively to evaluate the microinhomogeneity of deformation of metals independently of the chosen mesh size of the dividing grid. New structural characteristics are introduced: the base and wave of variation expressing the degree of crumbling or amalgamation of metal grains by inhomogeneity of deformation.Translated from Problemy Prochnosti, No. 7, pp. 40–44, July, 1991. 相似文献
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水三相点是开尔文热力学温度的唯一基准点,也是ITS-90国际温标重要的定义固定点。因此,水三相点不确定度分析对整个温标的建立、温度量值传递起着至关重要的作用。近3年来,中国计量科学研究院研制出一系列高质量的水三相点容器,加强了水三相点的研究,为不确定度的分析提供了更为可靠的实验依据。同时,不确定度的分析也是客观评价新研制容器性能的一个重要指标。因此,根据实验结果对新研制容器所复现的水三相点进行了不确定度评定。评定结果表明,其扩展不确定度为0.16mK(k=2.69,P=0.99)。 相似文献