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1.
在研究鸡法氏囊病毒(HBDV)的A片断cDNA结构的基础上,建立了RT-PCR早期诊断技术,结果表明,使用PrimerDesign引物设计软件,在VP5和VP2的重叠基因区设计的一对引物具有非常高的同源性和保守性,采用的四种IBDV的标准毒株,一种野毒株和一例病鸡标本通过此引物进行RT-PCR检测均得到了明显的阳性扩增结果,而两种其他鸡病病毒和一例健康鸡的法氏囊组织扩增均为阴性结果。  相似文献   

2.
李坏死环斑病毒检测方法的比较研究   总被引:1,自引:0,他引:1  
针对感染李坏死环斑病毒(Prunus necrotic ringspot virus,PNRSV)植株病毒的特异性检测,将DAS—ELISA、RT—PCR方法综合改进、优化,建立了免疫捕获反转录PCR(IC—RT—PCR)检测PNRSV的方法.该方法采用PNRSV抗体特异性免疫捕获病毒,并根据GenBank中PNRSV序列设计的特异性引物对PNS/PN3进行PCR扩增,得到约760bp大小的特异性基因片段.扩增产物经克隆测序后,结果表明:产物序列与PNRSV的外壳蛋白编码基因存在94%~98%的同源性.检测广泛性以及灵敏度比较研究显示,IC—RT—PCR方法能从含量很低的受病样品中检出PNRSV,检测灵敏度与RT—PCR相近,但所检测样品的范围更广,适用于从植物各部位检测低含量的病毒.IC—RT—PCR检测灵敏度比DAS—ELISA方法高出1000倍.  相似文献   

3.
目的 建立阿留申病病毒PCR检测方法,以用于实验动物雪貂阿留申病病毒的检测。方法 参考Genebank中阿留申病病毒核酸VP2基因序列设计一对引物,建立PCR检测方法,进行特异性、敏感性测试,并对实验雪貂粪便样品进行筛查。结果 测序结果显示,使用设计的引物可特异性地扩增获得阿留申病病毒的VP2中531 bp基因片段;在常见实验动物DNA病毒,小鼠多瘤病毒、小鼠细小病毒、犬细小病毒、疱疹病毒、鼠痘病毒、仙台病毒、小鼠腺病毒中均未扩增出目的条带;以目的片段核酸为模板测试该方法敏感性,检测下限达到90. 6 copy/μL。应用该方法对39份雪貂粪便样品进行检测,未检测出阿留申病病毒核酸阳性。结论 本研究建立的方法具备一定的特异性和敏感性,可作为实验动物雪貂中阿留申病病毒感染病原筛查的方法。  相似文献   

4.
RT—PCR检测草鱼呼肠孤病毒的方法研究   总被引:3,自引:0,他引:3  
草鱼呼肠孤病毒(Grass carp reovirus,GCRV)为草鱼出血病的病原.本实验根据GenBank中GCRV和其他水生呼肠孤病毒毒株的第六基因片段,在其保守区设计了一对GCRV特异性引物,建立了快速检测GCRV的逆转录聚合酶链式反应(RT—PCR)方法.该PCR体系中,上下游引物的最适终浓度为120nmol/L,最适退火温度为52℃.PCR特异性试验表明:所设计的引物只能扩增GCRV的核酸,而不能扩增嗜水气单胞菌BSK-10、WSSV以及正常CIK细胞的DNA或RNA.敏感性试验表明,当GCRV的反转录模板稀释至5-7时,PCR结果还为阳性.用所建立的RT—PCR方法对5份样品进行检测,结果表明本研究建立的RT—PCR检测方法可靠且可行.  相似文献   

5.
根据GenBank中已发表的3株WSSV的VP28基因序列,设计了4条特异性引物.通过筛选最佳反应条件和试剂工作浓度,建立了检测克氏原螯虾WSSV的巢式PCR方法.该方法第一轮扩增的敏感性是36ng,第二轮扩增的敏感性是36Pg.在对9份克氏原螯虾临床病料的巢式PCR检测中,用引物W1和W2做第一轮扩增,未检测到阳性病例;用引物S1和S2做第二轮扩增,检测到一例WSSV阳性病例.以上结果表明:对于检测克氏原螯虾WSSV,巢式PCR较一步PCR具有更高的检测灵敏度和应用前景.  相似文献   

6.
目的建立检测实验猴及生物制品中猴空泡病毒(SV40)的PCR方法。方法根据SV40-776株设计合成三对引物对现有毒株进行PCR扩增并克隆测序。用这三对引物对56份猴肾、肺、脾及10批脊髓灰质炎疫苗进行检测。结果三对引物均扩增出目的片段;56份猴脏器和10批疫苗SV40检测均为阴性。结论56只恒河猴和10批疫苗中未检测到SV40,所设计的三对引物检测结果准确,均可用于检测。  相似文献   

7.
利用RT—PCR方法从铜绿微囊藻PCC7806 mcy基因中扩增出目的基因片段,用TA克隆方法将基因片段插入pGEMT—T Easy中来制备质粒标准品.通过不同连续稀释倍数的质粒标准品成功构建了16srRNA和mcyH两基因的标准曲线,该法构建的两基因标准曲线r^2分别为0.9916和0.9938,且质粒标准品具有较好的重复性,满足实时荧光定量RT—PCR的要求.  相似文献   

8.
在研究鸡法氏囊病毒(IBDV)的A片断cDNA结构的基础上,建立了RT-PCR早期诊断技术.结果表明,使用PrimerDesign引物设计软件,在VP5和VP2的重叠基因区设计的一对引物具有非常高的同源性和保守性.采用的四种IBDV的标准毒株、一种野毒株和一例病鸡标本通过此引物进行RT-PCR检测均得到了明显的阳性扩增结果.而两种其他鸡病病毒和一例健康鸡的法氏囊组织扩增均为阴性结果.  相似文献   

9.
鹅常见病毒性疾病的快速诊断技术的建立及其应用   总被引:4,自引:0,他引:4  
根据3种病原基因组相关基因的序列特征设计了三对PCR引物,用于鹅常见的病毒性疾病包括鹅副粘病毒病、鸭瘟、小鹅瘟的鉴别诊断.三对引物均能从各自的参考毒株扩增出与预计片段大小一致的产物,而对其它的病原的扩增结果均为阴性;应用建立的PCR方法对来自广西不同地方的38份临床可疑病例分别进行诊断,DPV、APMV-1和GPV的检测阳性率分别为66.6%(10/15)、55.6%(15/27)和75%(6/8),二重及三重混合感染的占42.8%(9/21).研究结果表明建立的PCR方法具有快速、敏感、特异的特点,可用于疫病的临床快速鉴别诊断特别是多重感染的诊断以及流行病学的调查研究.  相似文献   

10.
提取BHK21细胞增殖的亚洲一型口蹄疫病毒(foot-and-mouth disease virus Serotype Asial)强毒株YNAs1.1的RNA,用一对引物P7,P13经反转录(RT)-PCR法扩增了约674bp的DNA片段。克隆目的基因后,采用双脱氧DNA链末端终止法测得了YNAs1.1的VP1基因36-633核苷酸序列。分析表明,病毒VP1基因的核苷酸序列与以色列以及印度已报道的Asia1型FMDV的同源性分别为82.11%与88.07%,对应的氨基酸序列同源性为87.94%与93.47%。该序列在GeneBank登陆号为AF241566。  相似文献   

11.
The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3~4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰~ -7.8‰, PDB) and matrix dolomites (-6.6‰~ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084~0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078~0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.  相似文献   

12.
AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…  相似文献   

13.
本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。  相似文献   

14.
Various applications relevant to the exciton dynamics,such as the organic solar cell,the large-area organic light-emitting diodes and the thermoelectricity,are operating under temperature gradient.The potential abnormal behavior of the exicton dynamics driven by the temperature difference may affect the efficiency and performance of the corresponding devices.In the above situations,the exciton dynamics under temperature difference is mixed with  相似文献   

15.
The elongation method,originally proposed by Imamura was further developed for many years in our group.As a method towards O(N)with high efficiency and high accuracy for any dimensional systems.This treatment designed for one-dimensional(ID)polymers is now available for three-dimensional(3D)systems,but geometry optimization is now possible only for 1D-systems.As an approach toward post-Hartree-Fock,it was also extended to  相似文献   

16.
17.
The explosive growth of the Internet and database applications has driven database to be more scalable and available, and able to support on-line scaling without interrupting service. To support more client's queries without downtime and degrading the response time, more nodes have to be scaled up while the database is running. This paper presents the overview of scalable and available database that satisfies the above characteristics. And we propose a novel on-line scaling method. Our method improves the existing on-line scaling method for fast response time and higher throughputs. Our proposed method reduces unnecessary network use, i.e. , we decrease the number of data copy by reusing the backup data. Also, our on-line scaling operation can be processed parallel by selecting adequate nodes as new node. Our performance study shows that our method results in significant reduction in data copy time.  相似文献   

18.
R-Tree is a good structure for spatial searching. But in this indexing structure,either the sequence of nodes in the same level or sequence of traveling these nodes when queries are made is random. Since the possibility that the object appears in different MBR which have the same parents node is different, if we make the subnode who has the most possibility be traveled first, the time cost will be decreased in most of the cases. In some case, the possibility of a point belong to a rectangle will shows direct proportion with the size of the rectangle. But this conclusion is based on an assumption that the objects are symmetrically distributing in the area and this assumption is not always coming into existence. Now we found a more direct parameter to scale the possibility and made a little change on the structure of R-tree, to increase the possibility of founding the satisfying answer in the front sub trees. We names this structure probability based arranged R-tree (PBAR-tree).  相似文献   

19.
There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.  相似文献   

20.
The geographic information service is enabled by the advancements in general Web service technology and the focused efforts of the OGC in defining XML-based Web GIS service. Based on these models, this paper addresses the issue of services chaining,the process of combining or pipelining results from several interoperable GIS Web Services to create a customized solution. This paper presents a mediated chaining architecture in which a specific service takes responsibility for performing the process that describes a service chain. We designed the Spatial Information Process Language (SIPL) for dynamic modeling and describing the service chain, also a prototype of the Spatial Information Process Execution Engine (SIPEE) is implemented for executing processes written in SIPL. Discussion of measures to improve the functionality and performance of such system will be included.  相似文献   

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